University of Nigeria

Research Publications
Author

OKPALANMA, Emeka Felix

PG/M.Sc/88/6772
Production of Malt-Based Syrups from Sorghum
Title

(Sorghum Bicolor) and Millet (Pennisetum

Typhoiodes) Grains
Department Faculty

Agriculture

Food Science & Technology

Date

May, 1991
Signature
I ~ ~ U C T I OOF
N MALT-BASED SYRUPS FROMSORGHUM(SORGHUMBICOLOR)
AND MILLET (PENNISEWM TYPHOJDES ) GRAINS

OKPALANMA, EMEKA FELIX

PG/M .sc./88/6772 '

DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY

FACULTY OF AGRICULTURE,
UNIVERSITY OF NIGERIA,
iJSL'KKA.

I N PARTIAL FULFUS/1ENT OF THE REQUIREMENTS FOR

THE AWARD OF MASTER OF SCIENCE DEGREE (M,SC,) I N
FOOD SCIENCE AND TECHN'OLOGY,
UNIVERSITY OF NIGERIA, NSUKKA

MAY, 1991 .
Mr. Okpalamm, Emeka F e l i x , a postgraduate student i n
t h e Department of ~ ' o o dScience and Technology, and with
r e g i s t r a t i o n Number PG/M,~o/88/6772has s a t i s f a c t o r i l y
completed t h e requirements f o r course and research w r k
f o r t h e degree of master of s c i e n c e i n Food Science and
Technology, The work embodied i n t h i s t h e s i s i s o r f g i n d
and h a s n o t been submitted i n p a r t o r f u l l f o r any o t h e r
diploma o r degree of t h i s o r any other u n i v e ~ s i t y ,

t
/&JkhM
Dr. C.C.1 OlQUOHA DR. A e L . IIGXDRONYE
SUPERVISOR.
 DEDICATION

his thssis Is dedicated t o all widours

who could muster enough courage, patience

to s t a y and train their children properly

as my mother did.

r"
i
 iii.

ACKNOWLEDGEMENTS

I want t o express sincere g r a t i t u d e and ' d e e p s t a p p r e c i ~ l t i o nto

m. A. I. Ihekoronye, my S ~ p @ J X i s o rf,o r t h e many hours hg spent i n

b l p i n g m e plan, canplete and e v a l u a t e this research project. I also

extend my th&s t o all my lectu~cersi n the department of Food Science

tind Technology.
i
I g r a t e f u l l y acknowledge t h a s s i s t a n c e of t h e departmental

technologists .: Onuchukwu, Onyebuashi , Kalu and Nwokedi of ~ i o c h e m i s t r y

department.

I express my personal thanks to Rev. Fr. E u t a b a s f l i , M., Mr. Obadiegu,

M., Dr. Chinyere, P . and Kene Oranu f o r many things. Also to M r . J. Nwabue

f o r h i s patience and f o r t i t u d e i n typing the. manuscript.

F i h a l l y , I owe a s p e c i a l debt o f g r a t i t u d e to a l l my l e c t u r e r s i n

the University of I f e , during my f i r s t degree programme.

 V.

TABLE OF CONTENTS Page

TITLE PAGE

CERTIFICATION

DEDICATION iii
. .
ACKNOWLEDGEMENT

TIlBLE OF CONTENTS

LIST OF TABLES

LIST OF FIGURES vii

ABSTRACT viii
INTRODUCTION

LITERATURE REVIEW
Production of Sorghum and Millet; Grains
sorghum grain ..
Millet grain ..
..
C h e m i c a l and Biochemical s t u d i e s on Sorghum
and Millet g r a i n s
sorghun g r a i n ..
Millet g r a i n .. 'i

..
Malting' C h a r a c t e r i s t i c s of Nigerian Sorghum
and Millet VazLeties
sorghum malt ..
~ i l l e malt
t ..
~ l u c o s esyrups ..
k fi n i t i o n s ..
~e-s of Production of Glucose Syrups
~ c i d
conversion ..
Acid-enzyme conversion ..
Enzymeenzyme conversion 22
Refining .. 25 ,
Properties and Functional uses of Glucose
@..-..-\a ' C)C)
Malt Based Syrupm ..
Mtuihing method ..
Preparation of m a l t syrups

MATERIALS AND METHODS .

Materials ..
Source of c e r e a l g r a i n s ..
Methods ..
Determination of Malting Characteristics
of the Cereal G r a i n s , Sorghum and M i l l e t
Determination of moisture content
Determination of percentage foreign seeds
and braken kernels ,
Determination of thousand corn weight
Determination of getminative energy
Determination of germinative capacity
Det@rmination of Optimum Malting conditions
of t h e C e r e a l Grains ,,
Moisture Content as a function of s t e e p time
Determination of Optimum steep time
Determination of Optimum g e d n a t i o n period
Effects of k i l n i n g a t 4s0c and varying periods
of time on moisture con t e n t of the ma1 t.

..
Determination of malting l o s s e s as a function
of gemination periods
Production of Sorghum, M i l l e t Malts.
Evaluation of M a l t ' s q u a l i t y c h a r a c t e r i s t i c s
Determination of Cold Water e x t r a c t
Determination of d i a s t a t i c power
Dekermhation of ~ o water
t extract
Determination of malting l o s s
Studies on Malt's Amylases
Extraction of m a l t anylase
Preparation of 1% buffered s t a r c h s u b s t r a t e
 . vii.

3.5.3 Preparatioh of Maltose Calibration curve 43

3.5.4
.
Datemination of optimum p~ f o r amylase
activity 44
3.5.5 Oetenninationof Optimum temperature
f o r amylase a c t i v i t y .. 44
44
.
3.5-6 s t a r c h Extraction £ran t h e Cereal g r a i n s
3 7 ~ r o x h a t e / ~ h e m i c aAnalysis.
l of g r a i n s ,
m a l t t i and s t a r c h e s f r a n sorghum and m i l l e t . 47
3.7- 1 Crude P r o t e i n determination 47
3.7.2 F a t determination 48
3,.7.3 Crude fibre determination ' 49
3 ..7.4 ~ s Determination
h
3.7.5 Total Carbohydratedetennination
3,'7,6 ~ e l s t i n L z a t i o ntemperature determination 51
3.7.7 Starch determination in s t a r c h concantrate
by hydrolytic method. 53
3.8 Production of M a l t Based Syrups 54
3.8.1. wort Preparation by t h r e e stage'dacoction
mashing method from sorghum malt 54
E f f e c t of varying mash concentrations and
a a c c h a r i f i c a t i o n periods on reducing sugar
contents of worts i n a t h r e e s t a g e decoction
mashing, .. I 54
E f f e c t of varying concentrations of
glucomylase and s a c c h a r i f i c a t i o n periods
on the reducing sugar content of malt hydrolysates 55
M a l t Based Syrup Production 55 .
Glucose Syrup Production 56
Determination of some p r o p e r t i e s of syrups 57
~ e t e n n i n a t i o nof s p e c i f i c gravi ty/l)egtee
baume' 57

Percentage reducing sugar content/

Dextrose equivalent(IX1 value determination 58
Determination of Colour 60
CHAPTER 4. RESULTS AND DISCUSSION 61

4.1
sorghum and millet. ..
Malting c h a r a c t e r i s t i c s of the cereal grains,
61
Optimum malting conditions of ths cereal
grains, sorghum and m i l l e t 62
Evaluation of m a l t 1s q u a l i t y c h a r a c t e r i s t i c s 68
Determination of optimum pH and temperature
c o n d i t i o m f o r malt's amylase a c t i v i t y . 70
~roximate/chemical analysis of t h e sorghum/
m i l l e t g r a i n s and malts. , . 74
Chemical analyses of starches extracted
from millet and sorghum grains. 76
E f f e c t s of varying mash, glucoamylase concantra-
t t o n s and s a c c h a r i f i c a t i o n periods on r e d w i n g
sugar contents of the wort syrup. 77
Properties of malt based eyrups and acid
enzyme converted glucose syrups. 81
CHAPTER 5. SUMMARY AND CONaUSICbJS
. 84

REFERENCES
APPENDIX
 LIST OF TABLES

TABLE 1 - Proximete analyses o f . P e a r l m i l l i t ( ~ W I 3 ) (memanwith .

ranges i n Parenthesis where available) $0

- Sorghum m a l t proximate analysis a f t e r four days

1, 2
gemination . As
n 3 Checklist of properties and f u n c t i o n a l ' u s e s
of corn syrupta I n s p e c i f i c food products. 30

t* 4
Sorghum and millet .
Malting c h a r a c t e r i s t i c s of t h e cereal grains,
62

5 valuation of malt * s q u a l i t y charact@.ristics 70

@ 6 ~roximate/chemical analysis of grains and malts 75

n 7 Analyses of starches from millet and sorghum grains 77

8 Properties of m a l t based syrups 83 -

" 9
syrups. ..
Properties of acid-cenzyme converted glucose
83 4
 LIST OF FIGURES

A General manufacturing procedure f o r corn syrups 25

Properties and functional uses of corn syrups 31

production from c e r e a l g r a i n s ..
Flow c h a r t of w e t milling operations i n s t a r c h
46

Flow c h a r t of acid-enzyme converted glucose syrups

production em 56

Flowchart of malt based syrup production 57

~t'

time(hours) ..
P l o t of moisture contents(%) a g a i n s t steeping
63

P l o t of d i a s t a t i c power(o~)against steeping time

(hours) a f t e r 4 days of germination 64
P l o t of d i a s t a t i c power (OL) against gurmination
periods(days1 a f t e r Sohours of steeping . 66.

periods(days) .
P l o t of malting l o s s ( % ) against gemination
67

period(hours) a t 45 C. .
P l o t of moisture cogtent(%) against kilning
, 69

p l o t of mg maltose against p~ . 72
. .
P l o t of maltose c a l i b r a t i o n curve 7

p l o t of mg ma1t o s e against t e m p e r a t ~ r e ( ~ ~ ) \.73

p l o t of glucose standard curve 78

and time(hours1 ..
P l o t of mg dextrose against mash concentration(%)
79

P l o t of mg dextrose a g a i n s t ~ 1 ~ = c s - i - ~ ~ L ~ c
concentration(%) and time (hours)
 xi.
ABSTRACT

he s u i t a b i l i t y of two cereals(sorghum and millet) f o r the production

of malt-based slyrup w a s determined.

i'roximate a n a l y s i s waa c a r r i e d on the grains. The g r a i n s were steeped

f o r SO hours, germinated for 5 days a t room temperature and k i l n e d f o r 48hrs

a t 4s0c. Ma1t i n g c h a r a c t e r i s t i c s of t h e g r a i n s determined include: the

germinative energy, germinative c a p a c i t y , 'Hot water e x t r a c t and d i a s t a t i c

power. S t a r c h w a s e x t r a c t e d from the two g r a i n s and used f o r syrup production

Optimum c o n d i t i o n s f o r t h e a c t i o n of m a l t amylases in syrup production were

a l s o determined,

The ma1t gs q u a l i t y c h a r a c t e r i s t i c s analysed showed t h a t sorghum g r a i n

generated b e t t e r malt. Malted g a i n s contained higher amounts of p r o t e i n and

crude f i b r e , and lower amounts o f f a t , ash, and t o t a l carbohydrates than tha

h a l t e d gains. 1000 g r a i n s o f m i l l e t and sorghum weighed 6 - 8 and 33,3g

respectively. Malting l o s s v a l u e s w e r e 16-20% f o r m i l l e t and 12016% f o r

sorghum. Germinative c a p a c i t y of the millet g r a i n s was 85% while sorghum

g r a i n s had a germinative c a p a c i t y of 90%. Hot water e x t r a c t v a l u e s were

1 8 0 . 1 ~ ~ / fko~r m i l l e t and 2 0 ~ ~ ~ O~timumd i a s t a t i c power of

3 f o~r sorghum,

2 7 O ~and 3 2 O ~were obtained f o r m i l l e t and sorghum r e e p e c t i v e l y , Sorghum

s t a r c h y i e l d e d syrup of a b e t t e r quality than millet s t a r c h . Sorghum s t a r c h

a l s o has a lower g e l a t i n i z a t i o n temperature and lower ash c o n t e n t than

millet s t a r c h . Optimum pH range f o r alpha amylwe a c t i v i t y i n b o t h m a l t

e x t r a c t s was 6-7, Optimum tempera- range f o r anylase a c t i v i t y was

found t o be 40-50°c f o r m i l l e t and 60-70°c f o r sorghum.

 CHAPTER 1

INTRODUCTION

The development of ma1t based syrup i n v o l v e s t h r e e fundamental stages:

'(1) Production of m a l t by a process c a l l e d malting.

(ill P r e p a r a t i o n of wort from t h e m a l t by d e m t i d n mashing process.

(iii)F u r t h e r s a c c h a r i f i c a t i o n o f the w a r t to malt based ~ y r u pusing

e x t e r n a l microbial amylases. ,

Glucose syrup i s t r a d i t i o n a l l y produced from corn s t k c h , hence

its name 'Corn s y r u p r . I t i s t h e p u r i f i e d ' c o n c e n t r a t e d aqueous s o l u t i o n

of n u t r i t i v e s a c c h a r i d e s o f Dextrose equivalent(DE) 20 o r more obtained .

by h y d r o l y s i s of e d i b l e s t a r c h e s !Whistl.er a t ' al,, 1984). The h y d r o l y t i c ,
-1

agents i n c l u d e a c i d , microbial amylases, m a l t amylases and combinations of

these. F u r i a (1968) has d e s c r i b e d t h e technology of glucose syrup

production, while M ~ ;U&istgr(1979)

C has described t h e a c t i o n p a t t e r n s of

enzymes used i n t h e commercial c o r n syrup production.

Malting is e s s e n t i a l l y a b i o l o g i c a l process in which t h e germination

of cereal g r a i n i s c a r r i e d o u t i n a c o n t r o l l e d environment. The t e c h n i c a l l y

intportant f e a t u r e s of g e m i n a t i o n are t h e synth.gies o f h y d r o l y t i c enzymes

and t h e degradation o f t h e g r a i n structure. When both p r o c e s s e s have

reached t h e d e s i r e d s t a g e , t h e germination i s i n t e r r u p t e d by d r y i n g o r

kilning,
 Malting s t u d i e s have been done by s e v e r a l workers; Aisien(l983)

i n v e s t i g a t e d t h e u t i l i z a t i o n of s o l u b l e carbohydrates during sorghum

yern~indtion.. Novellie (1960) e s t a b l i s h e d t h a t sorghum ma1t do n o t posses

t h e b - c u n ~ l c i s e,protease, c e l l u l a s a and hemi$ellulase a c t i v i t i e s of e i g h t

c u l t i v a r s of process millets.

Decoction mashing t r a d i t i o n a l l y employs malt which i s less modified . -. . L

thm t h a t used i n i n f u s i o n mashing and i s only l i g h t l y kilned(Briggs.5: &,

w r i n g t h e k i l n i n g process, an optimum temperature is chosen t h a t

s t r i k e s a balance between t h e development.of the c h a r a c t e r i s t i c m a l t

flavour, c o l o u r and the sustenance of high ma1t d i a s t a t i c power. The

1

colour i s produced d u r i n g ' k i l n i n g through m a i l l a r d r e a c t i o n s between the

p r o t e i n s and s u g a r s p r e s e n t i n t h e m a l t , (a n a t u r a l browning r e a c t i o n ) ,

Malt based syrups are used widely i n t h e food i n d u s t r i e s :

( i ) Brewing industry: D i a s t a t i c syrup c o n t r i b u t e s t o converting o t b r

s t a r c h y a d j u n c t s to simpler s u g a r s while n o n - d i a s t a t i c m a l t syrup

c o n t r i b u t e s towards t o t a l fermentables.
(11) Malt syrups .are i n c r e a s i n g l y being used as n a t u r a l food c o l o u r a n t s

thereby replacing caramels,

(iii) I n t h e baking industry, D i a s t a t i c malt syrups may be used i n breads

a s a yedst food t o r e l e a s i n g sugars n a t u r a l l y and c o n t r i b u t e t o l o a f volume

irnd texture. Malt syrup is a l s o used i n brown bread and dark cake manufacture,

b r e a k f a s t c e r e a l ,and b i s c u i t manufacture .
(iv) I n t h c p h a n n a c e u t i c a l industry, m a l t syrup could be incorporated i n t o

i n f a n t l i q u i d drug mixtures a s sweetening, colouring and f l a v o u r carriers. -

G ~ U C O S syrup
~ is widely used i n confectlonary and baking i n d u s t r i e s ,

i n canning of f r u i t s and vegetables, s o f t drink i n d u s t r y , i n beverages, and

other products r e q u i r i n g sweetness. Hoover(1963) has i l l u s t r a t e d t h e

functional p r o p e r t i e s of corn syrups a s they r e l a t e b > t h e type of conversion,

f i g u r e '2 Hoover(1964) has a l s o prepared a c h e c k l i s t of p r o p e r t i e s and

f u n c t i o n a l u s e s of corn syrups in a wide v a r i e t y o f foods, (Table 9)

The value of N i g e r i a ' s annual consumption of glucose syrup was

estimated atsN80.625 m i l l i o n and t h a t of c r y s t a l l i n e glucose I n form of

dextrose monohydrate was estimated a t about W60 million(Federa1 I n s t i t u t e

of I n d u s t r i a l Research, Technical memorandum No, 25, 1970)- These d a t a

derived from t h e l i m i t e d market surveys conducted were more l i k e l y t o b e

an under e s t i m a t e s t h a n over-estimates.

I n Nigeria, t h e r e l a t i v e abundance o f Sorghum and millet crop8 w i t h

an average m u a l production f i g u r e of 4,8 metric tonnqr and 2.4 metric

t o m e s respectively(Sumraru* miscellanous paper 90, 1979), has prompted

the current research e f f o r t s towards c o s t reduction. I n 1982 alone,

Nigeria's import value f o r m a l t was p u t a t about M40 million i n foreign

exchmge. Both wheat and corn are today in Nigeria ,Golden cereals': EF!A,
i

hence the quest f o r s u b s t i t u t e and/or blends.

Malting of t h e l o c a l c e r e a l s , sorghum and millet, generate endogenous

malt amylases which augment the imported microbial amylaae used i n t h e .

sclccharification process of malt symp pr=2utticn, thereby saving cost.

A i m s and Objectives of t h e Study:

The study was c a r r i e d o u t t o prepare malt-based syrups from l o c a l l y

avail able c e r e a l grains. The s p e c i f i c objectives were:

1. t o determine which of the two c e r e a l s , sorghum and m i l l e t has a higher

ma1t i n g potential.

2. to determine
,
t h e malting conditions necessary f o r optimizing the

sorghum/ m i l l e t malts' d i a s t a t i c power.

3. to determine the l e v e l s of microbial ainylase and conditions s u i t a b l e

f o r t h e production o f r e l a t i v e l y cheap malt based syrup.

 CIWTER 2

2. LITERATURE REVIEW

2.1 Production of Sorghwn and M i l l e t grains:

2 . 1 Sorghum qrain:

h l though vorghum rank8 f o b # mong c e r e a l s in c u l t i v a t e d a r e a world

wide following wheat, r i c e , and maize, i t i s t h e most important cereal i n

Nigeria occupying about 46% of t h e total land area devoted t o t h e growing

of ceredls. The a r e a devoted t o sorghum has i n c r e a s e d by about 25% over

t h e l a s t two decades growing from 4.6 m i l l i o n h e c t a r e s i n 1959 t o qin

e ~ t i m a t e d6.1 m i l l i o n h e c t a r e s i n 1979. Sorghum p r e s e n t l y accounts f o r

about 50% of t h e t o t a l c e r e a l production i n t h e country. Production has

\
gone from 2.5 m i l l i o n metric tonnes i n 1960 to 4.8 m i l l i o n metric t o i a s

i n 1978(Samaru miscellaneous paper No. 90, 1979). I n 1981/82, production

was 16,192 tonnes and y i e l d p e r h e c t a r e was 841 kilograms (Federal o f f i c e
i
of s t a t i s t i c s , Lagos, 1983. Survey of modern Holdings of A g r i c u l t u r e

1981/82, p. 2 0 ) .

The o r i g i n of t h e c r o p was traced to o t h e r r e g i o n s of A f r i c a , from

E t h i o p i a a c r o s s t h e Sudan (Damon, 1962). . A survey of t h e indigenous

sorghum v a r i e t i e s r e v e a l f o u r economically important varieties name:ii,

I

Guinea, kaura, F a r a f a r a , and Chad r a c e s (Buntung and C u r t i s , 1970).

2.1.2 ~ i l l e grain:
t

Millet although n o t as important a s some of t h e o t h e r cereals when

t o t a l world production figures are considered >it is nevertheless t h e 6

 6.

- G.,
b a s i c d a i l y d i e t ' o f s e v e r a l m i l l i o n people i n A f r i c a and I n d i a ( ~ a r t i ne t

There i s an average annual production of 2.4 m i l l i o n metric t o m e s and

d n a t i o n a l average y i e l d of 75Okg/hd (Sanaru miscellanous paper, 90; 1979). -

1n'1981/82 production w a s 9,862 t o m e s , and y i e l d p e r h e c t a r e , 566 kilogram

( ~ e d e r a lO f f i c e of s t a t i s t i c s , Lagos, 1983). There are two main types of

millet grown i n Nigeria. These are t h e Gero and Maiwa types. The Gero

m i l l e t s are of s h o r t e r d u r a t i o n taJcing 75-100days t o mature. Maiwa on t h e

o t h e r hand t a k e s between 120-150 days t o mature. T k Gero type, because

of i t s g r e a t e r a d a p t a b i l i t y , i s favoured over maiwa and over 8C% of a l l

millet grown i n N i g e r i a i s of the Gero type(Samaru miscellanous paper,

NO', 90, 1979'). Rachie (1974 l i s t e d the canmon and corresponding s c i e n t i f i c

names of t e n v a r i e t i e s of millets grown world over.

2,2 chemical and Biochemical s t u d i e s on Sorghum and Millet grains:

2,2,1 Sorghum grain:

'Neucere and sumrell (19801, s t u d i e d t h e proximate a n a l y s i s , f a t t y

dcid composition, f r e e sugars, mineral c o n t e n t and t h e i i d i s t r i b u t i o n of

t a n n i n s i n f i v e v a r i e t i e s of sorghum biccdor(L). Moench, They showed t h a t

t h e v a r i e t y w i t h predanlnantly f l o u r y endosperm (NSA 740) has t h e h i g h e s t

p r o t e i n c o n t e n t , . Some d i f f e r e n c e s i n f a t , ash, carbohydrate, and , f i b r e -

c o n t e n t s w e r e a l s o noted among t h e f i v e v a r i e t i e s . The c o n t e n t o f n e u t r a l

l i p i d s i n t h e f i v e lines of g r a i n sorghum ranged from 2.66 t o 3.49%. They

a l s o noted s u b s t a n t i a l d i f f e r e n c e s in t h e mineral uptake of t h e f i v e

v a r i e t i e s and according t o t h e r e s u l t of comparative m a l y s i s of f i v e

sugars i n theee v a r i e t i e s s t u d i e d , n m e l y ; f r u c t o s e , glucose., sucrose,

m d l toae a d raf f h o s e , f r u c t o s e and glucose comprised t h e h i g h e s t c o n t e n t s

m
of f r e e sugdrs.

Rooney a d s u l l i n s (1970) i n t h e i r study, compared t h e g r a i n produced

on d i p l o i d ( 2 ~ )and t e t r a p l o i d (4x1 l i n e s of t h e sorghum, Sorghum b i c o l o r

(L) moench, c u l t i v a r T x 403 f o r p h y s i c a l , morphological and chemical

properties. ~ c c o r d i n gt o their r e s u l t s , g r a i n f r a n t e t r a p l o i d was

y r e d t e r i n k e r n e l size and p r o t e i n c o n t e n t and was lower i n s t a r c h c o n t e n t

and test ? e i g h t than g r a i n from t h e d i p l o i d s . Mean v a l u e s of d i p l o i d and

t e t r a p l o i d s were 12.8 and 15.x p r o t e i n , 72.3 and 68.8% s t a r c h , 26.8 and

41.4 g/1000 k e r n e l s , and 74.3 and 70.0kg/hl respectively. The.result

a l s o showed t h a t endosperm cells o f t h e t e t r a p l o i d were l a r g e r than t h o s e

of t h e d i p l o i d . Kernel d e n s i t y and amino a c i d composition were similar.

Hoseney et &. ,(1974 ) examined t h e s t r u c t u r e of sorghum g r a i n

samples by scanning e l e c t r o n microscopy. They observed t h a t t h e s o f t o r

opeque endosperm i s c h a r a c t e r i z e d by r e l a t h e l y l a r g e i n t e r g r a n u l a r air

spaces, and showed t h a t its s t a r c h was e s s e n t i a l l y round and covered w i t h

a t h i n sheet of p r o t e i n . Furthermore, they discovered t h a t t h e hard

endosperm r e s u l t e d from s t r o n g adhesion between p r o t e i n and s t a r c h and

a l s o when t h e hard endosperm was f r a c t u r e d , many a t a r c h g r a n u l e s were

broken r a t h e r t h k t h e s t a r c h p r o t e i n i n t e r f a c e being broken. Results

a l s o r e v e a l e d t h a t a dwarf v'ariety from Sudan. had r e l a t i v e l y few p r o t e i n

bodies i n t h e endospenn and t h a t amino acid a n a l y s i s confirmed t h a t this

v a r i e t y contained 3.019 l y s i n e p e r 1009 p r o t e i n , s i g n i f i c a n t l y more than

normal i n sorghum grain.
1

S u l l i n s and ROOney (1974')compared sorghum g r a i n s t h a t d i f f e r i n

endospenn t e x t u r e and endospenn type i n order t o e v a l u a t e t h e usefulness

of microscopy t o account f o r d i f f e r e n c e s observed i n t h e feeding p r o p e r t i e s

of t h e s e grains. They observed t h a t t h e waxy.aorghum

. . .
kernel sections

hdd the smdllevt proportion of p e r i p h e r a l endospenn a r e a of t h e four

g r a i n s excrmined. The waxy s e c t i o n s were a l s o more r a p i d l y s o l u b i l i z e d by

pronase and alpha- amylase enzymes and by buffered .rumen f l u i d than, the

non w a x y sections. According t o them, t h e findings'might account f o r

observcltions of feeding trias in which steers fed non-waxy sorghum g r a i n

d i e t s r e q u i r e 8 t o 20X more feed t o produce a pound of g r a i n than steer

f e d waxy sorghum g r a i n d i e t .

Deyo et &., (1990) determined t h e proximate and amino a c i d composition

of mature and immature samples of sorghum grain. Their data indicate

marked d i f f e r e n c e s in amino acid content. ~ h e iobserved t h a t crude

p r o t e i n c o n t e n t of immature and mature sorghum g r a i n was shilar. The

feeding s t u d i e s which they c a r r i e d o u t showed less a v a i l a b l e energy fram

immature than mature sorghum grain.

Haikerual and ca hie son( 1971) determined total p r o t e i n and amino

i

acid composition of a number ofsorghum sample including those from two

f i e l d experiments. They showed t h a t t h e germ contained ' t h e h i g h e s t proportion

of p r o t e i n , followed by the whole kerzel, t h e endosperm and t h e p e r i c a r p

a l s o t h a t t h e amino ,acid c a p o s i t i o n of those p a r t s was d i f f e r e n t , with

higher proportion of l y s i n e , h i s t i d i n e , a r g i n i n e , g l y c i n e , a s p & t i c a c i d ,

threonine, and v a l i n e i n t h e g e m , than t h e whole kernel.

2.2.2 M i l l e t grain;

Bcrdi et_ &.,(I9761 i n their work, found t h a t p e a r l millet s t a r c h

ranged i n diarneter from 8 - 12 11, somewhat smaller than c o r n o r sorghum

atarch. They observed t h a t p a s t i n g p r o p e r t i e s of m i l l e t s t a r c h were

e h i l a r to those of sorghum s t a r c h , e x c e p t during the 1 hour holding period

a t 9 5 O ~ . They showed t h a t m i l l e t s t a r c h contained 1%amylose compared

w i t h 23% i n sorghum s t a r c h . Arnylograms of m i l l e t f l o u r a l s o gave low peak

v i s c o s i t i e s compared t o sorghum f l o u r i n d i c a t j n g an a c t i v e alpha-anylase

4)
s y s tern.

proximate a n a l y s i s of millet g r a i n has been c a r r i e d out. Shepherd

-
et, - dl., (1972) r e p o r t e d from E a s t A f r i c a t h e proximate c o m p o s i t i o n o f

m i l l e t g r a i n ( ~ r weight
y basis). P r o t e i n ranged between 11.5 and 13.8%

l i p i d ranged f r m 4.8 - 9.2%, f i b r e 1.0 - 3.8% and ash, 1.1 - 2.4%.

Table 2.1, shows t h e proximate a n a l y s i s a s r e p o r t e d by some authors,

g e n e r a l l y , p r o t e i n (%I vary from 8.4 - 21.8, l i p i d ( % ) from 2.9 - 7.5,

carbohydrate (%) ' f r a n 53.9 - 83.8, F i b r e ( % ) from 1.2 - 10.7, a s h ('961, from

- --
Badi e t ' aL., (1976) showed t h a t p e a r l millet &aln endospenn was

composed of both h a r d ( t r a n s l u c e n t 1 and s o f t ( o p e q u e ) p a r t s . . The hard p a r t

h s t i g h t l y packed, polygonal shaped s t a r c h - granules and a m a t r i x , p r o t e i n

c o n t a i n i n g r e l a t i v e l y l a r g e , embedded p r o t e i n bodies. The s o f t endospenn

F e t u g a (1977) Africa
G a d r y and Bideau(1974) Africa
nagbail( 1977-~ersondl
comnunication to Hulse Africa
et -
- al.(1980)
P o p l i and Singh(l972) India
L'prety and A u s t i n (1972)
Nigeria
Range o f means
Range of r a n g e s
hds l o s e l y packed, s p h e r i c a l s t a r c h g r a n u l e s c ~ v e r e dw i t h a t h i n s h e e t

of p r o t e i h . The s o f t endosperm c o n t a i n s many a i r spaces, and no p r o t e i n

bodies.

Lorenz a d liinze ( 1976 determined a d c&npared t h e functi&nal

c h u a c t e r i s t i c s of p r o s and f c r x t a i l millet s t a r c h e s with those of wheat

and r y e s t a r c h e s . The m i l l e t starches showed higher water binding

c a p c i t y values and g e l a t b i z a t i o n temperatures than t h e wheat s t a r c h .

with two exceptions, t h e millet s t a r c h e s produced swelling power values

d t 90°c which were similar t o t h o s e of the wheat s t a r c h . They observed I

t h a t t h e s o l u b i l i t i e s of t h e millet s t a r c h e s were lower than t h o s e of t h e

wheat s t a r c h , e x c e p t f o r t h e s t a r c h from one v a r i e t y of m i l l e t ; and t h a t

t h e amylograph v i s c o s i t i e s of m i l l e t s t a r c h e s were higher than those of

t h e wheat s t a r c h a t a l l r e f e r e n c e points.

l t a s u l -e-~ ( 1 9 7 7 )s t u d i e d t h e weight and composition of m i l l e t p a r t s . He

found t h a t t h e seed c o a t s contained r e l a t i v e l y high percentage of p r o t e i n s ,

sugars, and f a t s . The seed c o a t s had high c o n t e n t s of pentosans,

ht?raicelluloses, and f i b r e which are s i g n s o'f low n u t r i t i v e value.

. Removal

of seed c o a t s from m i l l e t s l e d t o a h i g h e r n u t r i t i v e value.

-
Ramachandra e t ' &.,(1977), found t h a t t h e t g t a l phenol and t a n n i n

l e v e l s of f i n g e r millet v a r i e t i e s i n d i c a t e wide v a r i a t i o n s i n phenolic

contents. T-hey showed thatqwhite g r a i n v a r i e t i e s had lower phenolic

c o n t e n t t h a n , t h e brown-grain varieties. I n v i t r o protein digestibility

values of low tannin samples were h i g h e r than those of t h e high t a n n i n

simples. Dehulling had e f f e c t of removing most o f . t h e phenolics from

f inger , rnille t g r a l n w i t h .concomitant i n c r e a s e i n i n v i t r o p r o t e i n

digestibility.

- A.,
A U ~ U St et ( 1979) , analysed f o r p r a k h , SZL~Z
acid c = p ~ s i t i W ,

and mineral a s s a y of 14 inbreed l i n e s o f p e a r l millet(Pennisetum americanum

(L) ~ e a k e )from t h e p l a n t , b r e e d i n g . proyrdn a t T i f t o n , Georgia. Tkiir

d a t a shared t h a t p r o t e i n c o n t e n t v a r i e d ' f r o m 10.7 - 17.1%. Chemical

s c o r e s on the amino a c i d s showed l y s i n e t o be t h e l i m i t i n g amino acid.

hey e s t a b l i s h e d t h a t mineral c o n t e n t v a r i e d considerably among t h e

d i f f e r e n t hybrids. The predominant elements were phosphorus and potassium.

p r u t h i and ~ h a t i a ( l 9 7 0 )s t u d i e d two improved strains4of Pennisetmm - i

) were found t o have a l i p i d c o n t e n t o f about 5.0%

t y p h o i d e u n ( @ b a j r a @and

and bound l i p i d c o n t e n t of about 0.5%. They observed t h a t i n t h e non-polar

f r a c t i o n , s t e r o l esters, hydrocarbons, xd t r i g l y c e r i d e s , are t h e p r i n c i p a l

constituents. They s e p a r a t e d p o l a r l i p i d s by twodimentional t N n - l a y e r

chromdtography and l e c i t h i n was found to be t h e major component.

Dorisova et s.,(1982) i n v e s t i g a t e d t h e e f f e c t of v a r i o u s s t a g e s of

t h e t e c t h o l o g y of process m i l l e t on t h e amino a c i d c o n t e n t o f m i l l e t

protein. They s t a t e d t h a t t h e l e v e l s of methionine and t y r o s i n e in

husked m i l l e t i n c r e a s e d by about 1196, whereas those o f l y s i n e and g l y c i n e

decreased by 1% and 11%canpared to unhusked millet. P o l i s h i n g of

husked m i l l e t decreased t h e l e v e l of g l y c i n e by 22.- and t h e l e v e l s of

threonine, t y r o s i n e by 7.1% to 13.8% compared to unpolished husked m i l l e t .

Cooking of polished husked m i l l e t decreased t o t a l amino acid c o n t e n t by

2.3 ~ dt i n.g chdrac teristics of Nigerian sorghum and Millet v a r i e t i e s :

2.3.1 Sorghum malk

Aisian G,, g.,(1978) in a study of t h e germination behaviour of

Guinea corn, (Sorghum v u l g a r e ) i n v e s t i g a t e d its percentage germination

(germination energy) and l e n g t h of t h e a s c r o s p i r e ranged between 2-2.5cm,

1
he r e s u l t a l s o showed t h a t t h e optimum moisture c o n t e n i f o r r a p i d .

g e m i n a t i o n was between 35 and 40%, a t opkhum t e m p e r a t i r e of 22Oc. he

r e s u l t s f o r t h e rest of t h e germinative c a p c i t y , percentage germination a t

d i f f e r e n t times of g e m i n a t i o n are t a b u l a t e d .

Daiber and ~ o v e l l i e ( 1 9 6 8 )found t h a t g i b b e r a l l i c a c i d had l i t t l e e f f e c t

on amylase development in normal k a f f i r corn. They observed t h a t only

immature seeds and very l a r g e g r a i n s produced more amylase when t r e a t e d w i t h

g i b b e r a l l i c a c i d , - b u t this e f f e c t was much s m a l l e r than t h a t found w i t h

barley. They concluded on f u r t h e r i n v e s t i g a t i o n t h a t amylase ,formation i n

sorghum appears t o be preponderantly' a f u n c t i o n ' ,of t h e embryo.

~ o v e l l i e ( l 9 6 0 )e s t a b l i s h e d t h a t sorghum m a l t s are poor i n betzi-amylase

compared w i t h b a r l e y m a l t s ( b u t s i m i l a r t o Oat and r a g i ) and do n o t possess

i

high d i a s t a t i c power, H e showed t h a t sorghum m a l t s contained beta-ainylase

i n c o n s i d e r a b l e q u a n t i t i e s , 18-39% of Ule sdcchslri-k'ying r l c t i v i t y being due

t o t h e beta-amylase. H e found t h a t t h e alpha-and b e t a - amylases developed

a t approximately t h e same r a t e d u r i n g germination since t h e i r r a t i o ( w h i c h

vAries from 0.22:l to 0.64:1) was p r a c t i c a l l y c o n s t a n t throughout the

ma1t i n g process.

~ i s i e n ( 1 9 8 2 )i n h i s s t u d i e s found t h a t modification i n t h e sorghum

g r a i n endosperm during s e e d l i n g growth and malting was a s s o c i a t e d mainly

w i t h increased a c t i v i t i e s of alphsmylas4endo-b)-4lucanase, limit

dextrincise and endoprotease. H e found t h a t t h e major s t a r c h - degrading

enzyme was alpha-amylase and also observed t h a t t h e a c t i v i t i e s of endo-

/+gluccmase, l i m i t d e x t r i n a s e a d endoprotease were comparatively higher, in

t h e endosperm than in the embryo during s e e d l i n g growth.

~ d y l o r ( l Y 8 3 )i n h i s study, observed t h a t when sorghum i s malted, much

of t h e n i t r o g e n i n t h e k e r n e l i s t r a n s f e r e d t o t h e r o o t s and shoots. His

exanination of Osborn p r o t e i n f r a c t i o n s e x t r a c t e d from t h e k e r n e l r e v e a l s

t i n t h e c a s e of b a r l e y t h e prolamins are t h e m a j o r source of t h e

t t ~ ds

nitrogen transferred. E'urthennore, he fouhd t h a t t h e 'two most important

f r e e a c i d s of sorghum m a l t appear t o b e asparagine and glutamine, as i n

germinated wheat and maize.

w i l l i a m ( 1983) s t u d i e d t h e e f f e c t s of t a n n i n on malting a s we1.l a s the

change i n polyphenols during malting of b i r d - r e s i s t a n t and non bird-

resistant cultivars, H e observed t h a t no d i f f e r e n c e could be found i n

t h e p e r c e n t germination nor in the r o o t and s h o o t production of t h e m a l t s

of t h e two c u l t i v a r s , H e found t h a t t h e r e was an. i n c r e a s e i n t h e antho-

cyanidin c o n t e n t of t h e r o o t s and shoots during malting.

Nigerian s p e c i e s of sorghum
~waifotl983),9ma-irradi~~dtw0 -
Sorghum acaUdatum(sk. 5912) and sorghum guineense(HP - 3)
'9
p r i o r t o malting

on a c o b a l t i r r a d i a t o r . H e exposed t h e s p e c i e s to t h e following doses;

0.22, 0.44, 1.76, and 4.95 k r d - I n t h e a s s a y s f o r d i a s t a t i c power,

/+amylase, and a l p b a n y l - .W h i l e i n t h e assays f o r germinative energy

and l e n g t h s of r o o t l e t s and acrospire,, they were exposed t o a dose of

0-5 krd. H e found t h a t a dose of 1.76 k r d r a i s e d t h e d i a s t a t i c 'power,

6-mylase ,g(- dmylclse, germinative energy, and lenghts of r o o t l e t s and

crcrospire ~n(utimcrllyr e l a t i v e t o those of t h e u n i r r a d i a t e d sorghun is't%

species studied. H e observed t h a t the e f f e c t of 1.76krd was, however,

higher i n Sk 5912 species than i n HP3 species.

irnfchie( 1982) studied f i v e Nigerian sorghum ' v a r i e t i e s B.E.S., F.F.B.L.

~~21 9 , LRV, and came c u t with results of the proximate a n a l y s i s

~ ~, 1 4 9 and i

a f t e r four days of g e m i n a t i o n as shown on t a b l e 2.2 below,

TABLE , 2 .

SORGHUM MALT PROXIMATE ANALYSIS (ANICHIE 1982)

-- - -- -- - --

Ma1t i n g l o s s (%) 25,OO 24.00 22-00 22.50 17-82

log f i l t r a t i o n time 1.2b 2 .01 1.62 1.78 1.52

Ma1t Nitrogen (%I 1,70 1.68 1-61 1-54 1-74

Aisien (1982) i n v e s t i g a t e d t h e u t i l i z a t i o n of s o l u b l e carbohydrates

during sorghum germination and seedling growth, He determined sucrose,

r a f f i n o s e and f r u c t o s e l e v e l s in the s c u t e l l v n of i n t a c t and excised

sorghum seedling during growth, H e found t h a t i n the scutellum of the

i n t a c t g r a i n embryo, sucrose and r a f f i n o s e l e v e l s declined sharply over

the germination phase b u t increased a t post-germination (ie r o o t enwgence)

a s hexose sugars from the modifying endosperm passed into t h e scutellum.

tie observed t h a t maltose, m a l t o t x i o s e and glucose were t h e main products

of t h e enzymic modification of t h e endospem during s e e d l i n g development,

which is a post-germination e v e n t atxi t h e r e f o r e concluded t h a t t h e growing

-.---z cf t h e embryo, w i t h its higher i n v e r t a s e a c t i v i t y showed g r e a t e r

. --!

cdpdcity f o r sucrose metabolism than t h e scutellum.

2.3.2 Millet Malt:

I
opoku et &.,(1981) g e m i n a t e d m i l l e t g r a i n s f o r 84h and k i l n e d a t

45Oc t o o b t a i n a m a l t product. They conducted a n a l y s i s of vitamins,

phytate, oxillate, tannins, total phenols, and calcium t o d e t e r m i n e t h e

n u t r ' i t i o n a l value of t h e g r a i n s and the malt. hey found t h a t the.,.levels

of vitamins were higher i n t h e m a l t than i n t h e grains. Also t h a t s l i g h t

i n c r e a s e s i n p r o t e i n and tote$ phenol w e x e observed in t h e m a l t , while

l i p i d , phytase, and o x a l a t e l e v e l s decreased during malting. The r e s u l t

of t h e proximate a n a l y s i s w a s given i n a table. . '

Skovron and Lorenz (1979) determined t h e - a?ylase, p r o t e a s e ,

c e l l u l a s e , and hemicullulase a c t i v i t i e s of e i g h t c u l t i v a r of proso

( ~ a n i c u mmiliaceum) m i l l e t s . They found t h a t a l l t h e c u l t i v a r s showed

b-my1 ase, pro t e a s e , c e l l u l a s e and hemicellul a s e a c t i v i t i e s with the

exception of one sample that sllowed no hemicellulase a c t i v i t y . The

optimum pH f o r b-amylase a c t i v i t y was found t o b e approximately 5.0, and

production of maltose p e r in'illilitre of e x t r a c t ranged from 0.73 t o 1.93 fi

6
a f t e r l h of incubation a t pH 5.25. Also t h e pH optimum f o r p r o t e a s e

a c t i v i t y was n e a r 3.0 and 5.0, production of t y r o s i n e p e r m i l l i l i t r e of

e x t r a c t ranged f r m 12.5 to 75.5 Yj a f t e z ii! of h ~ & i i i i o i i ai pH 4.8.

 Opokus 3 d.,(1983) s t u d i e d t h e q u a n t i t d t i v e and q u c l l i t ~ t i v echanges

i n c d b o h y d r a t e s , p r o t e i n s , and l i p i d m a t e r i a l s d u r i n g t h e germination of -
millet. They found t h a t a two-stage metabolism was e x h i b i t e d d u r i n g

ycrmin~i
t i o n a d t h t ~Y t a c h c o n t e n t d e c r e a o d d u r i n g germination which

coincided with cir~ increacja i n s o l u b l e carbohydrate and p r t e i n s . . They

f u r t h e r observed t h a t tho high l i p i d c o n t e n t o f t h e g r a i n was reduced t o

Ueleia culd arts son-Varriano(198Ib) s t u d i e d t h e e f f e c t of p e a r l millet

dnylases on i n t a c t s t a r c h g r a n u l e s and heated s t a r c h suspensions, Amylases

i n crude m i l l e t e x t r a c t s showed higher amylolytic a c t i v i t y on wheat s t a r c h

than on m i l l e t s t a r c h , b o t h i n amylograph determination and s t u d i e s on

hydrolysii of raw s t a r c h e s . According to t h e r e s u l t s , t h e a c t i v i t y p a t t e r n

of m i l l e t alpha amylase was similar t o t h a t of o t h e r cereal a l p h a amylases

'. w i t h t h e r a t e of appearances of h y d r o l y s i s products being dependent on the

particular starch substrates,

Gudisevd e t - &., 1981) screened twelve v a r i e t i e s of

' ( sorghum( Sotghwn

b i c o l o r ) , 14 v a r i e t i e s of p e a r l millet(Pennisetwn t y p h o i d e m ) , 12 v a r i e t i e s

of s e t a r i a ( s e t a r i a italics), f o u r v a r i e t i e s of ragi(E1eucine coracana),

11 v i d e t i e s o f echinocloa m i l l e t (Echinocloa c o l o n a ) , 1 3 v a r i e t i e s of

proso (Panicium meliacem), 11 varieties of kodo ;, (Paspalum scorbiculatum),

did 11 v a r i e t i e s of miliare(Pani.ciwn m i l i a r e ) f o r i n h i b i t o r y a c t i v i t y

a g a i n s t human s a l i v a r y amylase, Echinocloa, proso, kodo and miliare had

no d e t e c t a b l e a c t i v i t y . Two s t r a i n s of sorghum and one s t r a i p of p e a r l

millet d i d n o t show .&-anylase i n h i b i t o r y a c t i v i t y . ~ lo tlh e r s e e d s had

 6
a c t i v i t y , t h e highest being observed i n sorghum. According t o t h e r e s u l t s ,

t h e i n h i b i t o r s were non-dialysable and were i n a c t i v a t e d by pepsin treatment.

Also s e t a r i a and 8orghum i n h i b i t o r s vere r o l a u v e i y t i n e n o i a b i l e compared

to r a g i and p e u l m i l l e t inhibitors.

Maileshi and Desikacha (1979) evaluated t h e malting p o t e n t i a l of high

yielding v a r i e t i e s of ragi(E1eusine coracana). Three were found t o be of

good malting v a r i e t i e s a s they possessed good germinative energy, high

amylase a c t i v i t y , with good y i e l d s of malted flour. Gemination conditions

were 24h .teeping and 72h germination a t 25-26O~.

s k o r a i n and ~ a g l e ( 1 9 7 3 )in evaluating the use of b a j r a o r p e a r l

m i l l e t f o r malting purposes, compared the b e t a amylase a c t i v i t y of b a j r a

and barley malts. They found t h a t b e t a amylase a c t i v i t y of germinated

b a j r a increased up to 30h and decreased up t o 72h., while t h a t of b a r l e y

?

increased continuously up to 72h. They t h e r e f o r e concluded t h a t i f b a j r a

i s t o be used f o r m a l t production, then s h o r t malting i s radvocated.

Abdul-Hassan and Varriano-Martson(1982) - s t u d i e d t h e amylolysis of .

p e a r l m i l l e t s t a r c h and its f r a c t i o n s by p e a r l m i l l e t alpha amylase.

Gemination r e s u l t e d in a 120 f o l d i n c r e a s e i n s p e c i f i c a c t i v i t y of t h e

enzyme over t h a t of t h e alpha amylase f r a n mature grain. Results showed

t h a t raw m i l l e t s t a r c h w a s r e s i s t a n t to a t t a c k by alpha amylase. Fran

germinated m i l l e t , a l s o amylase was r e a d i l y hydrolysed by p u r i f i e d millet

alpha my1 ase, while sane p o r t i o n s of m i l l e t anylopectin were hydrolysed

slowly by alpha amylase.

 t'd et A.,
( lgVl6
) found t h a t decreasing germination temperatures .
fruu 35 t o 2 5 O ~i n b a j r a and from 25 to 1 5 O ~i n b a r l e y , = e s u l t e d i n a '
s i g n i f i c a n t i n c r e a s e in t o t a l amylolytic a c t i v i t y a s w e l l as p r o t e o l y t i c

a c t i v i t y of green m a l t s prepared fram the two cereals.. They observed

t h d t t o t a l m y l o l y t i c a c t i v i t y was mainly due to b e t a amylase i n b o t h

mdltk4, more s o i n b a j r a malt. They suggested t h a t t h e germination a t low

temperdture l e a d s t o b e t t e r y i e l d s a s w e l l a s q u a l i t y o f malt.

Pokhryal e t --
a l . , (1977) s t u d i e d hybrids of p e a r l m i l l e t g r a i n s

(Pennisetum typhoides Linn(Brum) s t a p t .dnd tiubb), They examined hybrids

f o r t o t a l p r o t e i n c o n t e n t and amino a c i d s p e c t r a . They found t h a t p r o t e i n

values ranged from 11.0 - 14.7X. Lysine and threonine which are l i m i t i n g

amino a c i d s according t o chemical s c o r e showed range o f 2 .S6 - 3.46 and

1.99 - 2.44 g / 1 6 g ~ , respectively. ,

L'dl a d.,
( 1973) compared v a r i o u s p r o p e r t i e s of m a 1 t from b a j r a w i t h

t h a t of b a r l e y malt. They observed t h a t proximate a n a l y s i s r e s u l t s o f t h e

two m a l t s showed l i t t i e d i f f e r e n c e i n t h e i r canposition. Both b a j r a and '

b a r l e y m a l t s had comparable amylolytic a s w e l l as p r o t e o l y t i c a c t i v i t i e s .

According t o t h e r e s u l t , there w e r e v e r y few d i f f e r e n c e s i n enzymatic

physical p r o p e r t i e s of a good malt though i t developed a b i t t e r t a s t e

after a short t i m e .

2.4 Glucose Syrups:

Contdolled h y d r o l y s i s of s t a r c h w i t h a c i d , enzymes, o r combinations

of t h e s e y i e l d s s e v e r a l s t a r c h hydrolysates which include: glucose syrup,

rnaltodextrinu, high maltose syrup and high fructose corn syrups and t h e i r

sol i d s respectively .
s t a c h from corn, sorghum, m i l l e t , potato, tapioca and other p l a n t

sources are used i n producing those hydrolysates.

2.4.1 ~ einitions:
f

syrup(Corn syrup).
Gl~.~cose Is t h e p u r i f i e d concentrated aqueous

solution of n u t r i t i v e saccharides of DE 20 o r more'obtained by hydrolysis

of e d i b l e starch.

Maltodextrin: Is a mixture of purified n u t r i t i v e saccharides obtained

by hydrolysis of s t a r c h having a DE of l e s s than 20. r/

~ i g hMaltose syrup: as t h e name implies has a higher than normal

maltose content when compared t o other enzymatically produced sykps.

High fructose corn S~~UE(H.F.C,S): Is corn syrup produced with t h e

additional s t e p of enzymic conversion of a portion o f P.glucose t o D-

fructose .
Dextrose Equivalent(DE): Is an indication of t o t a l reducing sugars

calculated as D-glucose on a dry-weight basis. The DE value is inversely

r e l a t e d t o the degree of polymerisation,(DP).

~non,(1979)c l a s s i f i e d corn s y w p s according t o method o t conversion;

acid conversion, acid-enzyme conversions and enzyme-enzyme conversion.

2.5 ~ e t h o d sof production of Syrups:

~ ~ U C O S ~

2.5-1 Acid Conversion: Acid conversion process i s c a r r i e d out i n d: pressure

vessel termed a I c ~ n v e r t e r * ~ # t a r icsh mixed with water t o form a suspension

o r s l u r r y , containing 3040% dry starch. The required amount of d i l u t e acid,

usually about 0.12%, based on the weight of s t a r c h , i s added and the

tt.rnper*tu~.er d i s e d by l i v e stem to 140-1600~. The h e a t i n g continues f o r

12-20 minutes, The cooked o r g e l a t i n i z e d s t a r c h i s converted f i r s t t o t h e

higher polysdccharides. As t h e process proceeds, o t h e r s u g a r s are produced;

~ccording B e M i l l ~ ( 1 9 6 7 1 ,tlle #- D -(1 44 ) l i n k a g e s undergo

hydrolysis more e a s i l y than do the 4- U - (13 6 ) linkages. Furthermore,

linkages n e a r e r t h e non-reducing end of t h e s t a r c h polymer are hydrolysed

more r a p i d l y than bonds l o c a t e d in the polymer i n t e r i o r , r e s u l t i n g t h e r e f o r e

i n a random hydrolysis.

H a r ~ e y ~ ( 1 9 8 3observed
) t h a t a c i d h y d r o l y s i s of g r a i n products is

considered t o modify f a t t y and p r o t e i n c o n s t i t u e n t s , r e s u l t i n g i n o f f - ,

flavoured m a t e r i a l s . According t o . h i m , a c i d h y d r o l y s i s c o n t r i b u t e s t o

the }\reduction of miscellaneous sugar products t h a t in t u r n can c o n t r i b u t e

t o v a r i a b l e f l a v o u r and f e n n e n t a b i l i t y .

w i t t and Blythe (1976) i n v e s t i g a t e d t h e f e r m e n t a b i l i t y of m a l t worts

s u p & n e n t e d w i t h 35% a c i d - thinned and 35% enzyna thinned corn syrup

s o l i d s , respectively. Under p i l o t brewing c o n d i t i o n s , t h e worts c o n t a i n i n g

t h e acid-thinned syrup showed a slower fermentation r a t e ,

2.5.2 Acid-enzyme conversion:

~cid-enzyme converted corn syrups are produced by m e a n s of a two-

s t a g e hydrolysis. The f i r s t s t a g e ( l i q u e f a c t i o n ) is accomplished w i t h

a c i d , a s described above, and i t s e x t e n t i s determined by t h e d e s i r e d DE

value and carbohydrate cornposition of tine f i n i s h e d syrup. The second

s t a g e ( s a c c h a r i f i c i l t i o n ) i s c a r r i e d o u t by means of s t a r c h hydrolysing

enzymes, u s u a l l y O(-my l a s e , P-amylase and glucoamylase depending on

t h e r e q u i r e d t y p e of c o r n s y r u p s and its composition. he a c t i o n p a t t e r n s

 22.

of enzymes used i n canmercial corn aymp manufacture have been described

by Mac A l l i o t e r (1979).

he scid-enzyme process according to Ough (1962) tends t o eliminate

carbohydrates degradation products and b-linked reversion products such

as gentiobiose. Hurst and Turner(1964) have described a patented process

for production of highly fermentable, n o n - ~ ~ ~ ~ t & ! , I i z icorn

; ; g syrdps with

high l e v e l s of glucose and maltose contents, w1th.a mixture of gluco-

cirnyldse and fungal d-amylasg . Different r a t i o s of P.glucose to maltose

can be obtained by a l t e r i n g the proportdo& of th. two enzymes, then

concentrations and conversion time.

Alternatively, when high maltose syrups a r e desired, barley &amylase

i s added and the hydrolysis proceeded u n t i l the required l e v e l of maltose

i s produced. Maeda and Tsao( 1979) reported the use of microbial 8-amylase
*
i n Japan i r r d u s t r i d l y r a t h e r than the p l a n t enzyme. Mltsushashi e t al.,

(1974) developed a patent which employs simultaneously, maltorgenic enyme

and pullulanase ( o(-l,6-glucosidasei irr tie prepuration of high maltose

syrups from acid l i q u i f i e d starch.

2.5.3 Enzyme-enzyme conversionr

High conversion hydrolysates are prepared almost exclusively by the

use of eniymes. Mac ~ l l i s t e r ( 1 9 7 9 )observed t h a t acid-catalyzed

hydrolysis of s t a r c h i s n o t capable of giving p r a c t i c a l hydrolysates with

more than about 90% P g l u c o s e , owing t o acid catalyzed reversion and

dehydration r e a c t i o n s r e s u l t i n g in a s i z e a b l e l o s s of D-glucose.
 The objective of t h e l i q u e f a ~ t i o nprocess is to convert a concentrabd

auepenoion of s t a r c h granules i n t o a s o l u t i o n of soluble d e x t r i n s of low

v i s c o s i t y f o r convenient handling in ordinary equipment and f o r easy

converoion to glucose by gluco-amylase. Mac Allister(1979) described t h e

process. According t o his process, a suspension of s t a r c h in water i s

treated with calcium hydroxide (slaked lime) to pH 6-7,q optimal f o r 0( -amylase.

Lime is used, because it serves a s a source of calcium ion needed by most

O( -amylase as a c t i v a t o r and s t a b i l i z e r . A s o l u t i o n of b a c t e r i a l M-amylase

i s then added, and the suspension i s pumped i n t o a steam jet where the

temperature i s r a i s e d inf&antaneoualy to 80-115~~. The s t a r c h is immediately

g e l a t i n i z e d a d i n the presence of t h e amylase, i s depolymerised rapidly to

- ilzld ;ass. Ir
The s a c c h a r i f i c a t i o n process t h a t follows ensures the conversion of I
II

s t a r c h t o D-glucose i n y i e l d s as high a s possible using glucoamylase. Once

the liquefaction s t a g e has'been completed, the r e s u l t i n g solution, containing

a mixture of maltose-oligosaccharides, is transformed to a high conwersion

syrup by holding i t f o r 36dOh in a stored tank a t appr6ximately SSOC and

pH 4.3 w i t h glucoamylase.

The amylt.r.se and amylopectin portions of s t a r c h are converted by

4 - a n y l d s e during liquefaction to a c o l l e c t i o n of l i n e a r and branched

dextrins. The l i n e a r d e x t r i n s are rapidly and almost . t o t a l l y converted

t o D-glucose by glucoamylase. The branched d e x t r i n s are much less

susceptible t o hydrolysis. A b d u l l a h e t &.,(1963) observed t h a t t h i s was

due t o the lower r a t e a t which glucoamylase cleaves the o ( - ( l j 6 ) D -

glucosidic linkage, as campared t o cleavage of theO(-~-(l+41 linkdge.
 Tkirpk &.,(1976), using a s i n g l e enzyme system, produced glucose

syrup m d dextrosd fran maize g r i t s . H e found o u t t h a t glucose syrup

production in. a s i n g l e enzyme e y s w with b a c t e r i a l 4-amylase a t pH 6.0

and 8 5 ' ~ eliminate many disadvantages of tkie d o a l e enzyme system eg.

microbial infection, pH adjustment during the reaction, high enzyme c o s t s ,

proteolysio. The syrup produced has a DE value of 38%. 0.3% ash and 0.03%

nitrogen.

Yoshizawaet &.,(1980) in their s t u d i e s found t h a t s t a r c h heated a t

1 2 0 f~o ~r 20 minutes was e a s i l y digested by o(-mylase a t p~ 6.0, while

raw s t a r c h was only p a r t i a l l y digested, Also they discovered that

d i & s t i b i l i t y of l i q u e f i e d corn s t a r c h was higher than t h a t of rice.

H i l r s t s &.,(1971) produced s t a r c h conversion syrups having a minimum

fermentable e x t r a c t s (F.E) value of 7%, a minimum dextrose e q u i v a l e n t ( ~ ~ 1

value of 47% and a max.dextrose content of 47% by s a c c h a r i f i c a t i o n of a

s t a r c h hydrolysate with an enzyme composition comprising a d i a s t a s e ,

glucoamylase and amylo -1,6- glucosidase.

-
Mandels et &,,(1975) reviewed the enzymic conversion of waste

c e l l u l o s e material t o glucose syrups f o r use in the food industry. They

discussed the production of a canplete c e l l u l a s e complex from Trichodenna

-Qu
Viride 9414 and pretreatment of s u b s t r a t e s by b a l l milling t o produce

maximum saccharification. Ac.cording t o the review, sane prunising s u b s t r a t e s

f o r conversion a r e listed: milled bagasse gave 42% s a c c h a r i f i c a t i o n in 4h,

milled m i l k cartons 81% saccharification i n 24h a t 50% pH 4.8.

Figure 2.1 shows a general manufacturing.procedure f o r glucose syrups.

 Process Ytee

Modern systems are continuous c o n v e r t e r s

F a t and protinaceous i m p u r i t i e s
precipitate.

Removes major p o r t i o n of i n s o l u b l e
impurities.
x

~ e m a i n i n gi n s o l u b l e i m p u r i t i e s removed

s o l i d s i n c r e a s e d t o 55%

&
/znzylne s a c c h a r i f i c a t i o n f For acid-enzyme hydrolysed syrups
enzyme t r e a t m e n t a p p l i e d a t this stage. .
I
47
Carbon r e f i n i n Powdered o r a c t i v a t e d g r a n u l a r carbon
used.

ion-exchange t r e a t m e n t is o p t i o n a l
used when ash free, very c o l o u r s t a b l e
syrup are desired.

S o l i d s i n c r e a s e s t o 82%

Fig. 1. A general manufacturing procedure f o r corn syrups.

2.6 Refininq:

I n t h e r e f i n i n g processes, h i g h q u a l i t y c o r n s y r u p s and s o l i d s o r
c r y s t a l l i n e d e x t r o s e demands t h e removal of:
( a! coloured compounds (b) m e t a l l i c ions.
(C) Hydroxymethyl f u r f u r a l

(dl nitrogen containing canpounds introduced with t h e o r i g i n a l s t a r c h

or with t h e enzyme preparations used i n t h e process

( a ) .Orgwic acids which can impart undesirable flavours o r colours to

t h e various products and

(f: sol-like p a r t i c l e s of unhydrolysed or degraded starch,

Carbon treatment removes most of t h e soluble proteinaceous material&

present and s u b s t a n t i a l l y a l l t h e 5-(hydroxy-methyl),- 2 - furaldehyde *

formed during the acid treatment, Also, many. commercidl-ly activated carbon

are e f f e c t i v e i n removal of heavy metals such as i r o n and copper,. t h a t

can a c t a s c a t a l y s t s f o r developing colour, - Most new ' i n s t a l l a t i o n observed

Conlee(l971) use counter c u r r e n t applicatiok of iactlvated granular arba an

in c y l i n d r i c a l column because it can be conveniently re-activated,

yielding more favourable economics,

A t y p i c a l ion-exchange deionization system c o n s i s t s of s i x fixed bed

columns (three p a i r s of c a t i o n and anion axcircuiyrt i.tz3in 03 aervice r'or

regeneration), The c a t i o n exchange r e s i n s used are strong acid exchangers

(sulfonated resins i n the hydrogen fonn) and the anion exchangers uaually

are weak base r e s i n s (tertiary m i n e i n the free base form), The anion

exchange r e s i n s removes acids generated by reaction of t h e , s a l t s i n the

syrup l i q u o r with t h e cation-exchange resins.

- &,(1981) discussed an experimental equipnent used f o r

Bezhal c t

e l e c t r o - f i l t r a t i o n of glucose syrups which achieves separation of various

i

foreign substeinces eg. micro-ozganims,pmteinls and o t h e r macromolecular

cornpoundrr, f u r t h e r colouring m a t t e r s , c o l l o i d s etc. The syrup p a s s

through a l a y e r of g r a n u l a t e d m a t e r i a l , under t h e a c t i o n of d i r e c t

elecuical c u r r e n t . The u n d e s i r a b l e p a r t i c l e s are coagulated and trapped

on t h e granules.

~ a l d a s s a r(i1971) described a process whereby concentrated solLtLons

o f sugar are t r e a t e d w i t h resins to remove i m p u r i t i e s such a s amino a c i d s ,

mineral a c i d s and salts, o r g a n i c b a s e s and a c i d s etc. by passage through

e s e r i e s o f f o u r ion-exchange r e s i n f i l t e r s . The r e s i n s are r e d m e r a t e d

w i t h 10-15% H2S04 o r 44% NaOH. I n c o n t r a s t w i t h o t h e r processes, t h i s

o n l y needs 2 4 o p e r a t i v e s , cuts i n d u s t r i a l costs by 80% and g i v e s an

improved pr@uc t.

Hersiczky (1972) c o n s t r u c t e d a f i l t r a t i o n u n i t f o r continous o p e r a t i o n ,

maximum c a p a c i t y 10,0001/h and s u i t a b l e f o r all t y p e s of r e a c t o r and

hydrolysis. With s i n g l e passage o p e r a t i o n , t h i s p u r i f i c a t i o n u n i t removed

94% of suspended m a t t e r f r a n upper, middle and lower r e g i o n s of the

hydrolysate and i n c r e a s e d o u t p u t o f the f i l t e r s t a t i o n by 50%.

2.7 P r o p e r t i e s and Functional u s e s of Glucose syrups.

'1
There are s e v e r a l types of c o r n syrups each of which has i t s own s a t

of p r o p e r t i e s . These p r o p e r t i e s are the sun of t h e c h a r a c t e r i s t i c s o f the

components which make up each syrup. These include: Dextrose e q u i v a l e n t

(DE), carbohydrate c a n p o s i t i o n , a c i d i t y and pH, sulphur d i o x i d e , fermentable

e x t r a c t , Baume and s p e c i f i c g r a v i t y , Ash, P r o t e i n , c o l o u r , v i s c o s i t y ,

Humectancy and hygroscopicity 'etc.

 ~ u d v i get &.,(1975), studied the f a c t o r s responsible f o r the browning

of glucose syrups during storage. They suggested t h a t i n i t i a l s t a r c h

materials must not contain 0,6 - Om% protein, with only small mounter

of o i l and fibres. Also after f i l t r a t i o n , a pre-concentration t o 28-32

t3dume' degrees i s necessary, followed by a second f i l t r a t i o n . Further more,

the optimal temperature f o r c r y s t a l l i z a t i o n i s important.

Keaslay ( 1978) studied the c a t a l y t i c hydrogenation of glucose syrups

as a means of controlling hygroscopicity and s u s c e p t i b i l i t y of browning

a d fermentation reactions without changing properties such a s v i s c o s i t y

9
osmotic p r e s s u r e or sweetness. According t o the r e s u l t s , hydrogenation

s i g n i f i c a n t l y decreased (P 0.05) moisture uptake of syrups a t 100% RHO

Browning decreased with DE before hydrogenation of syrups, and hydrogenation' .

(eg reduction i n DE from 100-76) of a syrup reduced colour development when .

the syrup was heated with amino acids. Furthermore, hydrogenation of

syrups decreased % fermentable sugars.

Hoover(1963) has i l l u s t r a t e d the functional properties of ,Corn Syrups

a s they r e l a t e t o tha type of conversion. This is shown i n f i g u r e 2.2.

he arrows of increasing s i z e p o i n t t o the d i r e c t i o n of t h e most d e s i r a b l e

corn syrup t o use f o r a p a r t i c u l a r property, a l l o t h e r f a c t o r s being t h e

same. I n s e l e c t i n g the most s u i t a b l e corn syrup, t h e g r e a t e r the number

of arrows t h a t go in the desired d i r e c t i o n , the b e t t e r is t h a t corn syrup

f o r t h e intended application. For example, i n choosing a corn syrup f o r f

I

use i n i c e cream production, the lower coriversion syrups are preferred.

These syrups.increaae the bodying and cohesive e f f e c t s , viscosity, and

prevent excessive growth of ice c r y s t a l s during freezing.

 29.

~ o o v e r ( 1 9 6 4 )has a l s o prepared a c h & l i s t of p r o p e r t i e s and

f u n c t i o n a l uses o f c o r n syrups i n a wide v a r i e t y of foods. Table 3 shows

t h e s e data. This table may b e used advantageously by f i r s t determining

t h e property o r p r o p e r t i e s of a food which may be improved w i t h t h e u s e

o f c o r n syrup and then s e l e c t i n g the most s u i t a b l e syrup. For example, in

bakery products higher conversion c o r n syrups are p r e f e r r e d where browning,

f ermen t a b i l i t y , sweetness, and f l a v o u r enhancement are d e s i r e d . '

2.8 Malt based Syrups.

2.8.1 Mashing Method.

~ a l its converted t o wort by b a s i c a l l y two methods of mashing; t h e

decoction mashing method and the single-temperature mashing system c a l l e d

i n f u s i o n method(Briggs e t a1 1981). I n i n f u s i o n m u h i n g process, no p a r t ' '

of t h e mash i s b o i l e d and r e t u r n e d to t h e main mash, r a t h e r t h e whole mash

i s g r a d u a l l y heated f r a n mashing-in t o rnashing-off. When the mashing-in

temperature ( 35-SOOC is p r o g r e s s i v e l y r a i s e d t o mashing-of f temperature

( 75-80°C through t h e s a c c h a r i f i c a t i o n temperature range of 65-70°c, thet

mashing process i s c a l l e d upward i n f u s i o n , while the downward i n f u s i o n

process resrllts when t h e f i n a l temperature o f t h e marsh(65-70°C) i s lower

than t h e i n i t i a l temperature o f t h e mash(75-80°c). ~ n f u s i o nmashing is

s u i t a b l e f o r ttie highly modified m a l t . (r"

I n decoctl-on mashing, ' a p o r t i o n of the mash i s b o i l e d and r e t u r n e d

to t h e rest of t h e mash in t h e mash tun. T r a d i t i o n a l decoction mashing

employs malt which is less modified than t h a t used i n i n f u s i o n mashing

--
and i s o n l y l i g h t l y k i l n e d ( 8 r i g g s e t a 1 1981). There are three d i f f e r e n t

k i n d s of decoction methods. '

 IYPF OF CORN S Y R U P '
PROPFRIY OR IIItJC1IC)NAI L J S F
- (ALPllAnEllCAILY) LOW.CONV. REG.-CONV. IN1ER.-CONV. IiIOII-CONV.

BODYING AGEN.1

BROWNING REACTION

CONFSIVENESS

FERMENTABII.ITY

FLAVOR ENIIAFICEMENT

FlAVOR TRANSFER
MEDIUM

FOAM STARILIZFR

)tVMECTANCY

blYGROSCOrlCITY

NUTRlllVE SOLIDS

OSMOTIC TRFSSURE

PRFVENTION 01: COARSE

I C E CRYSIALS DURING
FRtEZING

5HEEN PRODUCER
 Checklist of properties
and funclional uses o f corn syrups
in specific food producls

-- -. - -. . --- -. --- - .- -- -. -- -..--

-Baby -loods---- -- --- --- ---.- -
..-..Bakery
..... - producls
-. -. .............. - ..--.--..
-Beverages,
-..... -brewed
...... - ......-... -..-.
Bcveragcs, carbonated - lion alcoliulic
- .-.--- -- - ---- -- - - --. -
Breakfast loods
--------.---
---
Catsup, chili sauce, --
ton~dlo
-raucr
.- ...
- Cereals, prepared
0--- ----
- -
Cheese spreads and Irluds

-- Chewing
- gunr --
Chocolate p~oducls
-. -- - ... ...
Cilrus juices, .
dried
- ...... -.. ----.
,-.-

~ondcnsedmilk
.
-,
'
-Cunlrctionr - ---
Cordials and liqueu~s
.-... ........
Eggs, lruzer~or dried
.----
Extracts and llavors
-----------. --
- Frostings and icings
------ - ----- -
Fwil bullers
Fruit juicer and fruil jirice drinks
 ( i ) The one mash method'

( i l l The double mash method

(iii)The t h r e e mash methud.

K a r e l (1967) described and i l l u s t r a t e d t h e production, on a continuous

b a a i e a 'complete mash' c o n t a i n i n g almost a l l t h e s o l u b l e m a l t substances

and moat of t h e enzymes. I n h i s study, f i n e l y ground m a l t was mashed a t 5

6 5 O ~ ( 11 5 ) by s t i r r i n g f o r 30 seconds. The mash was then s u b j e c t e d t o a

p r e s s u r e shock of 294 p.s.i and immediately converted by passing through a .

p r e s s u r e r e l i e f valve i n t o a s a c c h a r i f i e r , where t h e temperature was i n c r e a s e d

by IOC p e r minute from '65O t o 70-71°c. T h i s temperature was maintained

u n t i l t h e i o d i n e test gave a yellow colour. The t o t a l conversion period

w a s 10-20 minutes,

. B a r r e t and G r i f f i t h s (1966) s t u d i e d s h e e f f e c t s of m a l t k i l n i n g on

wort p r o p e r t i e s , The r e s u l t s s h o w that as c o l o u r i n c r e a s e d and moisture

decreased t h e e x t r a c t value o f the m a l t s remained e s s e n t i a l l y c o n s t a n t

I
while t h e d i a s t a t i c power decreased s u b s t a n t i a l l y . ~ h l si n t u r n was

p a r a l l e l e d by a r e d u c t i o n of f e r m e n t a b i l i t y i n the d e r i v e d worts. Measurement

of t h e i n d i v i d u a l s u g a r s p r e s e n t i n t h e worts showed t h a t t h e decrease i n

f e r m e n t a b i l i t y was a s s o c i a t e d w i t h a diminution in the percentage of maltose

and an i n c r e a s e 10 t h e d e x t r i n c o n t e n t , while the values f o r o t h e r s u g a r s

were r e l a t i v e l y unaffected.

Desrousseaux and Montreuil(l966) s t u d i e s on commercial mashing showed

t h a t , /+amylase a c t i o n occured o p t i m a l l y a t about 6 3 O ~ , it slowed down as

t h e temperature reached 70°c, and w a s i n h i b i t e d a t higher temperatures. The

-
optimum f o r 1hit d e x t r i n a s e a c t i v i t y w a s 50-63O~, h i g h e r t e m p e r a w e s
destroying t h e enzyme. Alpha-amylase a c t i o n began above 63Oc, a t t a i n e d a

maximum around 72Oc and diminished a t higher temperature.

Narzisa and L i tzenburger (1977) , inves'tigated t h e mashing conditions

and gum contents and thus concluded t h a t it was p o s s i b l e to r e g u l a t e gum

contents t o some e x t e n t by v a r i e t l o n of mashing method b u t t h e s t a t e of

modlficetion of the malt plays a much more decisive p a r t than mashing

conditions. Also they added t h a t the a l t e r a t i o n of t h e pH of mashing t o

5.5 e f f e c t e d advantageous degradation of ti-glucan only a t high mashing

temperatures.

2.8.2 Preparation of m a l t syrups.

The t h r e e mash mmthod involves usshing i n a t about 35-40°c.

a After I

sometime about one t h i r d of the mash, t h e f i r s t mash, is taken l n t o a k e t t l e

I
1
and boiled and brought back to t h e mash tun where the temperature of t h e
i
i
whole mash i s r a i s e d t o 5 0 - 5 5 ~ ~t,h i s i s k e p t f o r a period of t h e , about
II
15-30 minutes. Boiling destroys t h e enzyme i n t h e boiled portion as
i

w e l l a s g e l a t i n i z e s t h e starch. D i a s t a t i c action is thus f a c i l i t a t e d by

1
t h i s process. Then a second mash (again one t h i r d portion of t h e e n t i r e

mash) i s boiled and returned to t h e main mash. T h i s brings t h e temperature

j
t o 6 0 - 6 5 ~ ~t h, e s a c c h a r i f i c a t i o n temperature. It i s allowed a t t h i s

temperature f o r 30-60 minutes. L a s t l y a t h i r d is t r e a t e d i n the same manner,

and i t r a i s e s tho temperature of t h e whole mash t o 70-75O~, the mashing-off

temperature. This i s a l s o k e p t f o r 30-60 minutes. 3

The r e s u l t i n g wort is c l a r i f i e d , theapH adjusted t o 4-5 and appropriate

amount of glycoamylases added. The s a c c h a r i f i c a t i o n is allowed to c o n t h u e

f o r several hours (12-72h) depending on t h e e x t e n t of conversion d e s i r e d

r~ldt h i s i s determined by i t s dextrose equivalent value(DE).

he melt syrup which contains a mixture o f saccharides i s neutralised

and concentrated by evaporation in multi-effect e v a p o r a b a or f o r

laboratory works, on a b o i l i n g water-bath to about 80% slolids s o aa t o

I

i n h i b i t microbial spoilage.
 CHAPTER 3

MATERIAIS AND METHODS

3.1 MATERIALS

3.1,1 M i l l e t g r d n s (Pennisetun~~ y p h o i d e s )and sorghum grains(s0rghwn

h l c o l o r ) were purchased f ran Orba m a r k e t , NsulEka.

3.1.2 ~myloglucosidase(AMG), from ~ s p e r g i l l u sn i g e r was purchased from

Nove ~ n d u s t t i a lEnzymes Division, Novo Alle DK - 2880 Bagsvaerd, Denmark.

I
I
I
3.2.3 Other Chemicals and r e a g e n t s were o f the p u r e s t a n a l y t i c a l grades.

3.2 -
METHODS

Determination o f m a l t i n g ~ h ~ w a c t e r i s t i cofs t h e c e r e a l q r a l n s ;
392 I

Sorqhum and m i l l e b ,

-j 2 .I Determination of moisture content:

I
The moisture c o n t e n t s of the g r a i n s and m a l t s w e r e determined i
I
i
&cording t o t h e I n s t i t u t e of Brewery, I,O.B.(1977) method of a n a l y s i s , !
dS ~ O ~ ~ O W S :

About 209 sample of g r a i n s were f i n e l y ground in a Thanas Wiley M i l l

Model ED-5, and thoroughly mixed. 5g of the ground sample was placed

in a moisture d i s h which was c l o s e d and weighed immediately to 0.001g.

i1 .
The cover of t h e d i s h w a s removed and placed in a pre-heated oven f o r I

e x a c t l y 3h a t I O S ~ C , The l i d was r e p l a c e d and removed 'from t h e oven,

then allowed t o cool i n a d e s i c a t o r f o r a t l e a s t 20 minutes t o ,room

temperature. The d i s h was t h e n re-weighed t o 0.001g.

Calculation: %

The moisture percentage(M) of the sample

Wkre W1 - weight of sample b e f o r e d r y i n g

3.2r2'.
w2 - weight of sample a f t e r d r y i n g

~ e t e r m i n a t i o nof Percentage Foreign Seeds and broken k e r n e l s .

The percentage of f o r e i g n s e e d s and broken k e r n e l s of t h e c e r e a l

g r a i n s were determined according t o t h e method of Association of O f f i c i a l

Analytical Chemists, A.OoA.Cm(1980) a s follows:

509 of each of t h e g r a i n s was weighed and the f o r e i g n s e e d s and broken

kernels were counted out. he g r a i n s were reweighed and t h e d i f f e r e n c e i n

weight recorded a s a percentage o f t h e o r i g i n a l weight.

3.2.3 ~ e t e r m i n a t i o nof thousand Corn weight:

A thousand corn weight of t h e g r a i n samples were determined according to

t h e method of l.O.B(l977) a s follows:

20g samples were weighed o u t a f t e r removal of f o r e i g n m a t t e r and half

corns. The number of c o r n s i n each sample counted and moisture c o n t e n t

determined. ;4

Calculation:

-The weight o f 1000 c o r n s o f d r y c o r n s i n gram(g)

Where W - t o t a l weight of c e r e a l g r a i n s taken

DM a D r y matter percentage of t h e g r a i n s

N = T o t a l number of c o r n s counted.
 I

3.204 a~
The o b j e c t i v e of t h i s test was to measure the. piercen t a g e of g r a i n s .

which cm be expected to germinate f u l l y i f t h e sample i s malted normally s

d t t h e time of t h i s test. The I,O,Ei.(1977) method of a n a l y s i s was adopted

as follows:

100 c o r n s f r a n the samples w e r e placed i n a p e t r i d i s h l i n e d w i t h two

f i l t e r pcrpers i n t h e bottom t o which 4 m l of water had been added, The

p e t r i d i s h was covered and t h e g r a i n s allowed t o g e m i n a t e i n a cupboard,

The c h i t t e d c o r n s w e r e removed a t 24, 48, and 72h from t h e beginning of

steeping,

Percentage of c o r n s chii2ted a s t h e g e q l n a t i v e energy were c a l c u l a t e d

thus:

Germinative energy = GE(%)

3.2 ,5 Determination of germinative Capacf t .

The o b j e c t i v e of tNs test w a s t o measure the percentage of l i v i n g

c o r n s i n t h e sample. The germinative cap'acity of the g r a i n s was determined

using the hydrogen peroxide method a s d e s c r i b e d by H o u g h s , &,,(1981)

a s follows:

200 c o r n s were s t e e p e d i n 2OOml of 0.75% hydrogen p e r o x i d e ( ~ ~ f0o~r

48h a t room temperature, The s t e e p l i q u o r w a s replaced w i t h f r e s h hydrogen

peroxide ' s o l u t i o n and was l e f t f o r f u r t h e r 24h. G e r n l i n a t i m c a p a c i t y was

then c a l c u l a t e d a s f o l l w s :

Where n I number of c o r n s t h a t d i d n o t g e h i n a t e ,
 3.3 ~kterminsCiotrof Optinrun Malting Conditions of the Cereal Grains:
?
.. .
? .I
A t u r e Content as a f u n c t i o n of s t e e p time:
i~iois

E i g h t p e t r i d i s h e s l i n e d w i t h filter papers a t t h e i r boktoms were ,

provided and f i l l e d with equal volumes of t a p water. 20g of t h e g r a i n s were

cledned m d steeped u t roOm temperature i n each of t h e p e t r i d i s h e s f o r

v x i o n s t i m e s , 10-80h, w i t h e i g h t hourly change of s t e e p l i q u o r . .

A t t h e end of edch s t e e p period, t h e g r a i n s were d r a i n e d , s u r f a c e

wdter b l o t t e d with f i l t e r paper, then t h e moisture c o n t e n t determined, as

i n section 3 - 2 ' 1

3.3.2 ~ e t e r m i n a t i o nof Opbhum Steep t i m e :

ha g r a i n s (209) were s t e e p e d a t v a r i o u s times, 10-80h as described

dove. Each of t h e e i g h t sets w a s allowed t o g e m i n d i e f o r 4 days i n a

deck cupboard and then k i l n e d f o r 48h a t 55Oc, after which t h e m a l t ' s

d i a s t a t i c power was determined as i n s e c t i o n 3.2 -4

3.3.3 Determination of o p t h u m germination period:

The g r a i n s (20g) were s t e e p e d f o r 50h and germinated f o r v a r i o u s

p e r i o d s , 1-7 days i n a dark cupboard, l a t e r k i l n e d f o r 48h a t 5 5 ' ~ and

t h e ma1 t m
s d i a s t a t i c power determined.

3.3.4 E f f e c t s of k i l n i n g a t 4 5 ' ~ and varying p e r i o d s of time on moisture

c o n t e n t of t h e m a l t :
- %

Samples of malted g r a i n s a t optimum malting conditions(50h steeping,

and 5 days germination) were Kilned a t v a r i o u s p e r i o d s ( l 2 h , 24h, 36h, 48h,

60h) a t 45'~. and moisture c o n t e n t determined.

 3.3.5 Determination of Maltinq l o s s e s a s a f u n c t i o n of germination periodr

The g r a i n s (209) were s t e e p e d f o r 50h and germinated f o r y a r i o u s

p e r i o d s , 1-7 days, then tho r e s u l t i n g malting losses p e r n t h day o f

t-
germination determined as i n s e c t i o n 3 -4-4 .

Production of swghwn, Millet Malts:

I k g of each of t h e g r a i n s was cleaned and steeped i n o r d i n a r y t a p

water f o r 50 hours a t room temperature w i t h 8 hourly change of s t e e p

l i q u o r t o both minimize t h e growth of microbes and p r o v i s i o n of more

oxygen t o t h e embryo of the g r a i n s . A t t h e end of t h i s s t e e p i n g p e r i o d ,

t h e g r a i n s were drained and spread on a cleaned f l o o r of a dark cupboard.

Wdter W ~ Ss p r i n k l e d on t h e c o r n s when t h e y were v i s i b l y d r i e d , t o e n s u r e

adequate moisture supply throughout t h e 5 day genningtion period.

Germin.ation was terminated by k i l n i n g a t 4 5 O ~f o r 48h in a hot-air

oven. A t t h e e x p i r a t i o n of t h i s time, t h e malt became f r i a b l e and t h e

k i l n i n g was stopped. .. ., .

30 4 - Evaluation o f Ma1t @
s quality characteristics:

3.4-1 ~ e t e r m i n a t i o no f Cqld Water Extract(CwE)

c o l d water e x t r a c t s of t h e m a l t s were determined according t o t h e

I.O.B.(1977) methods of a n a l y s i s as follows:

10y ground m a l t was d i g e s t e d w i t h 200ml of d i s t i l l e d water c o n t a i n i n g

121111of 0 . l ~ ammonia f o r 3h a t 20°c, s t i r r i n g d t h a l f Hourly i n t e r v a l s .

The r e s u l t i n g s o l u t i o n was f i l t e r e d and the s p e c i f i c g r a v i t y o f . t h e

0
f i l t r a t e measured a t 20 C O
Cdculation;

The Cold water e x t r a c t (CWE) %

x 20

where G r t h a excess degrees of g r a v i t y of t h e f i l t r a t e t a k i n g water a t

?oOc ds 1000.

ie G = 1000(SG - 1).

3.4.2 Determindtion of d i a s t a t i c Power (Using F e h l h q ' s T i t r a t i o n ) .

D i a s t a t i c Power determination w a s c a r r i e d i n accordance w i t h t h e

~ n s t i t u t eof Brewery, I.O.B. (1977) methods of a n a l y s i s a s follows:

iul u u f i l t e r e d c o l d water e x t r a c t o f a m a l t i n f u s i o n was prepared and

allowed t o ' s e t t l e . 3ml a l i q u o t o r s u i t a b l e volume of t h e s u p e r n a t a n t

l i q u i d was p i p e t t e d into 3 100ml of 2% b u f f e r e d s t a r c h s o l u t i o n attemperated

dt 20°c, and contained in 2 0 h l f l a s k . The f l a s k was shaken and maintained

d t t h i s temperature f o r e m c t l y 1 hour from when t h e al.iquot was added.

30ml of 0 . l N NaOH s o l u t i o n was added .to s t o p t h e r e a c t i o n , and made

up t o 2001111 a t 20°c w i t h d i s t i l l e d w a t e r . 5ml o f mixed Fehlings s o l u t i o n

was p i p e t t e d I n t o a 1501111 narrow-necked b o i l i n g f l a s k . The ,digested

s t a r c h s o l u t i o n was added from a b u r e t t e t o t h e c o l d F e h l i n g s t o within

l r n l of t h e f i n a l end point. T@ f l a s k c o n t e n t s was mixed and b o i l e d w i t h

moderate e b u l l i t i o n f o r 2 minutes. The b o i l i n g was continued and w i t h i n

1 minute, 3 drops ,of methylene b l u e i n d i c a t o r was added and t h e t i t r a t i o n

completed.

The end p o i n t was i n d i c a t e d by d e c o l o r i z a t i o n ' of t h o h e t h y l e n e b l u e

and t h e r e a c t i o n l i q u i d j u s t becoming' red.

C d c u l atlon:

Dias t a t i c power( DP) expressed i n degree ~ i n t n e r ( O ~ )

Where X = no of m l of malt e x t r a c t

y I no of m l of converted s t a r c h t o reduce 5ml o f Fehlingee.

S = t i t r e f o r s t a r c h blank.

Determination of t i t r e f o r s t a r c h blank:

The u n d i l u t e d 2% s t a r c h s o l u t i o n was t i t r a t e d a g a i n s t a mixture of

l m l of mixed Fehlinges s o l u t i o n and 2ml o f F e N i n g e s s o l u t i o n B, using

t h e technique described under method, w i t h methylene b l u e i n d i c a t o r .

(The blank may be neglected i f i t i s less than 3% of t h e measured

d i a s t a t i c value of t h e m a l t ) .

3-4.3 ~ e t e r m i n a t i o no f Hot Water E x t r a c t ( H w ~ ) r

The h o t water Extract of t h e m a l t w a s determined by t h e procedure

described i n t h o method o f I n s t i t u t e o r Brewery I.O.B(l977) as follows:

50g o f ground m a l t was mixed w i t h 360ml of d i s t i l l e d water p r e v i o u s l y

0
heated t o about 68 C s o as t o ensure an i n i t i a l mash mix temperature o f

6 5 O ~w i t h c o n t i n o u s ' s t i r r i n g f o r 10 minutes. The mixture was l e f t a t

6s0c ' f o r 1 hour. The mixture was t h e n q u i c k l y cooled t o 20°c(with ice

c h i p s ) and t h e volume m d e up t o 515ml w i t h d i s t i l l e d water, The mixture

was f i l t e r e d and t h e s p e c i f i c g r a v i t y of 'the f i l t r a t e was determined a t

20°c w i t h s p e c i f i c g r a v i t y k o t t l e w i t h i n one hour of c o l l e c t i n g t h e sample.

 Calculation of Hot W a b r Extract.

~ h t i~ x t r a c t ( E ) as-is' expressed a s l i t r e degrees/kg I G x 10.13.

Where (i - 0
excess degrees of g r a v i t y of t h e f i l t r a t e taking water a t 20 C

3 .4 -4 ~ e t e r m i n a t i o nof Malting Loss(%):

The percentage malting l o s s of t h e malted samples was determined

according to the method described by ~ o v e l l i e ( 1 9 6 2 )q$ follows:

A thousand kernel weight of t h e o r i g i n a l (unmal.kd) grain was

determined on a dry weight b a s i s before malting. After malting,. the

thousand kernel weight of the r 1 1 ~ 1 i ; e c isanple was a l s o determined after

removal of the roots and shoots by hand-threshing and moisture by heating

( d r y weight b a s i s ) .

Calculations:

Malting l o s s (%) = 100(Co Cn)-

co

where, Co - 1000-kernel weight of t h e m a l t e d grain.

cn P 1000 - kernel w t , of t h e m a l t on the n t h day of germination.

. .5 s t u d i e s on Malt's amylase:

3.5 ~ x t r a c t i o nof malt amylase:

,
Malt anyrase w a s extracted according t o t h e mthod of Shanbe 'et. - 2..
(1988) a s follows;

The malted grains (5.0g) (under optimum conditions) were ground

separately i n a morter, and q u a n t i t a t i v e l y transfered i n t o 100ml standard

volumetric f l a s k by washhg with d i s t i l l e d water, then made up t o 100ml.

 ~t was incubated a t 3 7 ' ~ f o r 3h i n 250ml c o n i c a l f l a s k and 2.01111 samples

withdrdwn, c e n t r i f u g e d (8000g, 0.5h) and t h e s u p e r n a t a n t s t o r e d i n a

r e f r i g e r &or.

3.5.2 P r e p a r a t i o n of 1;Yo b u f f e r e d s t a r c h s u b s t r a t e :

59 s t a r c h ( d r y b a s i s ) were made i n t o a p a s t e with a l i t t l e c o l d

w a t e r and then poured i n t o 400ml o f b o i l i n g water, The mixture w a s b o i l e d

f o r 2 minutes and then cooled. Adequate quantity o f each of t h e b u f f e r

s o l u t i o n s prepared ealier $ s e e appendix) w a s added and each o f t h e mixtures

was made up t o 500m1 r e s u l t i n g t o 1% b u f f e r e d s t a r c h s u b s t r a t e s o l u t i o n o f

pH 4, 5 , 6 , 7 and 8 r e s p e c t i v e l y .

3.5 - 3 erepclration of m a 1 tom c a l . i b r a t i o n curve:

A series of maltose s o l u t i o n s were prepared, s o t h a t 2ml c o n t a i n

0.4 - 2,Omg anhydrous maltose as follows:

I n t o e a c h of t h e t e n test tubes were added 0.4 - 2.0ml o f stock

standard maltose s o l u t i o n c o n t a i n i n g 2mg/2ml r e s p e c t i v e l y . The s o l u t i o n s

were r e s p e c t i v e l y made up to 2ml each by a d d i t i o n of a p p r o p r i a t e amount of

d i s t i l l e d water. 1 m l of t h e 1% s t a r c h s o l u t i o n and 2ml o f DNS r e a g e n t

were added.

The t e s t - t u b e s were t r a n s f e r r e d t o a rack i n a b o i l i n g water b a t h

and heated f o r fi;e minutes and then cooled t o room temperature a f t e r

which t h s c o n t e n t of each tube w a s d i l u t e d t3 2 0 m l w i t h d i s t i l l e d water.

., s u i t a b l e amount of each sample was poured i n t o a ' c o l o r i m e t e r cuve.tte f o r

o p t i c a l d e n s i t y determindtion a t 505 n'!n a g h a s t a r e f e r e n c e blank which

.-contain o n l y 2 m l water, Iml s t a r c h and 2ml DNS reagent.

'i'
 3 5 .A ~ e t e r m i n a t i o no f optimum p~ f o r amylase a c t i v i t s

2ml of t h e d i l u t e d amylase e x t r a c t (2ml e x t r a c t i n 2001111 d i s t i l l e d

water) was added to test tubes i n a rack-containing lml each of t h e 1%

buffered s t a r c h s u b s t r a t e s o l u t i o n a t t h e pH 4, 5 , 6 , 7, 8 previously

prepared. The tubes were shaken f o r 5 minutes to mix it properly and

i n c u b a t e d i n a t h e r m o s t a t i c a l l y c o n t r o l l e d water b a t h a t 37Oc f o r 10 minutes.

The d i a s t d t i c r e a c t i o n was stopped by t h e a d d i t i o n of 2ml D N S c o l o u r .

reagent. ~ l th
l e tubes were heated i n a b o i l i n g water b a t h f o r 5 minutes

and then cooled t o room temperature a f t e r which t h e . c o n t e n t s o f t h e tubes

were d i l u t e d t o 20ml w i t h d i s t i l l e d water, The absorbance was read a t i

505 run a g a i n s t a r e f e r e n c e blank. The blank was prepared by b o i l i n g t h e

arnylase e x t r a c t f o r 5 minutes b e f o r e adding to t h e 1%b u f f e r e d s t a r c h

substrate solutions, The c o n c e n t r a t i o n of t h e reducing s u g a r s a s maltose

i n t h e s t a r c h h ~ d r o l y s a t ewas c a l c u l a t e d by e x t r a p o l a t i n g i t s absorbance

value from t h e maltose c a l i b r a t i o n curve.

3.5.5 l a t e r m i n a t i o n of optimum temperature f o r amylase a c t i v i t g

2ml of t h e d i l u t e amylase e x t r a c t was added t o 1 m l of 1%s t a r c h

( s u b s t r a t e ) s o l u t i o n buffered a t optimum p~ range 6-7, and incubated f o r

30 minutes a t v a r i o u s temperatures, 30, 40, 50, 60, 70 and 80Oc.

;!ml of (D*) c o l o u r r e a g e n t was added(and o t h e r procedures r e p e a t e d )

and t h e c o n c e n t r a t i o n of reducing s u g a r s a s maltose c a l c u l a t e d .

3.5.6 s t a r c h e x t r a c t i o n from t h e c e r e a l m a i n s :

The w e t m i l l i n g method o f Watson(l97Q) was adopted i n t h i s work. The

I process involves c l e a n i n g , s t e e p i n g , c o a r s e m i l l i n g , deyerrning fine

~ r d l l i n y , s e p a r a t i o n of f i b r e , s t a r c h s e p a r a t i o n f r a n g l u t e n , and d r y i n g of

3tdrch as shown in the flow c h a r t ( F i g . 3.1).

Two kilograms of cleaned g r a i n was placed i n a troqgh, and covered

't
with 3 l i t r e s of 0.45% sodium m e t a b i s u l p h i t e s o l u t i o n . The trough was k e p t

i n a water b a t h maintained a t 4 5 O ~f o r 40h. A t the end of t h e 40h, t h e

g r a i n s were d r a i n e d and washed w i t h c l e a n water. The g r a i n was c o a r s e l y

ground w i t h a manually operated corona handmill, c o n s i s t i n g of two s t u d

p l a t e s of wtdch one p l a t e r o t a t e s while t h e o t h e r i s s t a t i c . The a d j a c e n t

f ~ e ;iif tl-ii plates are studded 'with t e e t h , which are s o arranged t h a t t h e

g r a i n s pass i n between t h e s t a t i o n a r y and moving p l a t e s . By a d j u s t i n g t h e

p l a t e s , it was p o s s i b l e to vary t h e d i s t a n c e between t h e t e e t h so a s to :

o b t a i n a d e s i r e d p a r t i c l e size. The purpose of this m i l l i n g s t e p is t o

t e a r the kernels apart i n order t o l i b e r a t e the gems. .Some:endosperm

s t d r c h was however l i b e r a t e d during degenning. \

The s l u r r y of c o a r s e l y ground g r i s t w a s degenned by f l o a t a t i o n i n a

b i g b a s i n of water. The f l o a w d g e m was scooped w i t h a guaze while t h e

rest of t h e m a t e r i a l s s e t t l e d . The o p e r a t i o n was r e p e a t e d many times by

s w i r l i n g t h e c o n t e n t o f t h e b a s i n u n t i l a l l the geqn w a s v i r t u a l l y removed.

The d e g e m d g r i s t was n e x t screened on nylon b o l t i n g c l o t h of 100 mesh to

remove t h e bulk of t h e f r e e , s t a r c h and gluten. The s c r e e n t a i l i n g f r a c t i o n

composed l a r g e l y of p i e c e s o f honey endosperm and f i b r e .

The s c r e e n t a i l i n g f r a c t i o n was f i n e l y ground to achieve maximuril

starch liberation. The f i b m w a s s e p a r a t e d from t h e s t a r c h and g l u t e n on

, \
 I--$
screenin

~ i g .3. Flow c h a s t of w e t m i l l i n g o p e r a t i o n s i n s t a r c h production from

cereal grains. . &/"
t h e nylon b o l t i n g c l o t h . .The starch a n d - g l u t e n f r a c t i o n s obtained from

both c o a r s e m i l l i n g and f i n e m i l l i n g were mixed. The s e p a r a t i o n of s t a r c h

.-
from g l u t e n was achieved by sedimentation. The s t a r c h with a d e n s i t y of . -.

1.5 s e t t l e d i n t h e aquaous s l u r r y a t a f a s t e r r a t e than t h e g l u t e n p a r t i c l e s

\
of d e n s i t y 1.1 (Kent, 1975)., T i m s e t t l e d g l u t e n and s t a r c h were made to

flow down a slop1)ed tray. The g l u t e n flowed o u t f i r s t . The . s t a r c h

f r d c t l o n wds f u r t h e r washed with water and t h e remaining glugen separated.

h f ' a i r l y gluten-free s t a r c h was obtained.

The s t i x c h c o n c e n t r a t e was placed between f o l d s of f i l t e r paper a d

0
dried i n an oven k e p t a t 50 C f o r 48h.
G

3*7 - ~ ~ r o a i m a t e / ~ h e m i caanla l y s i s of g r a i n s , ma1 ts and s t a r c h e s from

Sorghum and ~ i l l e t :

3.7,1 Crude p r o t e i n determination:

Crude p r o t e i n of t h e samples was determined i n accordance w i t h t h e

procedure of Association of O f f i c i a l A n a l y t i c a l Chemists A.O.A.C.(1980)

as follows:

bout 0.29 of t h e s a p l e w a s weighed o u t a c c u r a t e l y i n t o a 50ml

kjeldahl flask. The following were then added i n t o t h e f l a s k . 59

&hydrous sodium s u l p h a t e , 1g hydrated c u p r i c s u l p h a t e and l O m l concentrated

s u l p h u r i c acid. The d i g e s t i o n w a s c a r r i e d o u t by h e a t i n g t h e f l a s k on an

e l e c t r i c c o i l u n t i l i t s c o n t e n t s becane clear. Heating however was

continued f o r a t l e a s t one hour after t h e s o l u t i o n had c l e a r e d . ~ f t e r

kwating,the c o n t e n t s was t r a n s f e r r e d w i t h s e v e r a l washing i n t o 25Oml

v o l u n ~ e t r i cf l a s k , and was made up t o t h e mdrk a f t e r cooling.

D i s t i l l a t i o n apparatuswas s e t up, and steam was passed through it

f o r 10 minutes. 5rnl of b o r i c a c i d i n d i c a t o r was placed i n 25Qml c o n i c a l

fldsk. The c o n i d a l f l a s k was placed under t h e condenser such t h a t t h e

condenser t i p was placed i n the d i s t i l l a t i o n apparatus and was r i n s e d

 'i"
down w i t h d i s t i l l e d water.' The cup was c l o s e d w i t h t h e rod, and 5ml of

60% N&H was p u t i n ; t h i s was l e t In c a r e f u l l y , leaving 'behind a l i t t l e

to prevent ammonia escaping.Steam w a s t h e n l e t through f o r about 5 minutes

. -
( u n t i l t h e amount of l i q u i d in the c o n i c a l f l a s k was about twice what i t w a s i n

i n t h e beginning of d i s t i l l a t i o n ) . Then t h e b o r i c acid i n d i c a t o r was t i t r a t e d

w i t h 0.0W H c l t o the end point. The t i t r e was t h e number of m l s of 0,OlM

[icl trhdt changes t h e i n d i c a t o r from green t o p i n k i s h colour.

Calcul &ions:
-- ---

Let w r e p r e s e n t weight of sample

I,ut T r e p r e s e n t ~ n l sof titre 0.0l.M ~ c l

1 litreof MHC1 a 14.01gN

1 l i t r e of 0.0lM H c l = 0.4401 g~

Tml of 0.01M H c ~ = 0.0001401 x T g N

250
---1
5
of d i g e s t = 0.0001401 x T x 250gN -
5

% crude p r o t e i n = 0.0001401 x T x 250 x 6.25 x 100

, wx 5

3,7,2 F a t Determination:

F a t w a s determined according t o t h e A.O.A.C.(1980) method of a r d y s i s

as follows:
0
A n e x t r a c t i o n f l a s k was cleaned, d r i e d i n an oven a t 100 C and its

weight determined. 29 of t h e sample was a c c u r a t e l y weighed and t r a n s f e r r e d

I

i n t o t h e e x t r a c t o r thimble. The thimble w i t h the sample was then p u t into

t h e Soxhlet extractor. About three q u a r t e r s of, t h e c o n t a i n e r w a s f i l l e d

w i t h petroleum e t h e r .

The f l a s k was p l a c e d on t h e h e a t e r , t h e S o x h l e t e x t r a c t o r connected

to it and condenser i n t u r n connected t o t h e Soxhlet. The t a p to t h e

condenser was t u r n e d on and t h e h e a t e r switched on. The e x t r a c t i o n was

allowed to r u n f o r about 3h.

~t t h e end of t h e e x t r a c t i o n , t h e thimble w a s removed and t h e e t h e r

recovered. F i n d l y , t h e o i l wils dried a t 1 0 0 i~n ~an oven and t h e

e x t r a c t e d o i l weighed.

~alculatiom-

% fat L W t of o i l x
Wt. o f sample

3.7 , 3 Crude F i b r e determination:

The A m e r i c a n A s s o c i a t i o n o f C e r e a l c h e n i s t s , A.~.C.c.,(1976) method

o f a n a l y s i s wds employed i n the d e t e r m i n a t i o n o f c r u d e f i b r e i n t h e samples

a s follows:
ti'
The r e s i d u e s from e'ther e x t r a c t d e t e r m i n a t i o n or d e - f a t t e d samples

were t r a n s f e r r e d t o a d i g e s t i o n f l a s k . 200ml of b o i l i n g 1.5% s u l p h u r i c

a c i d was added. The d i g e s t i o n f l a s k was t h e n connected to a condenser and

heated, The f l a s k was f r e q u e n t l y r o t a t e d u n t i l t h e s a n p l e w a s thoroughly

wet. The f l a s k was removed after 30 minutes, f i l t e r e d through l i n e n i n

a f u n n e l and washed w i t h b o i l i n g w a t e r u n t i l the f i l t r a t e was n o l o n g e r

acidic. The i n s o l u b l e matter was washed back i n t o the d i g e s t i o n f l a s k

c o n t a i n i n g b o i l i n g 15% NaOH s o l u t i o n . The f l a s k was connected to a

r e f l u x condenser and b o i l e d f o r 30 minutes, a f t e r which t h e m i x t u r e was

 SO.

d l o w e d t o s t m d f o r b o n e minute. f he c o n t e n t s were then f i l t e r e d through

a cheese c l o t h i n a funnel and residue thoroughly washed w i t h b o i l i n g water

II

and then w i t h 1%hydrochloric a c i d , and again w i t h b o i l i n g water u n t i l

no l o n y e r a c i d i c . Then i t w a s wcrshed twice w i t h 95% e t h a n o l , t h r e e times

w i t h d i e t h y l e t h e r , and f i n a l l y t r a n f e r r e d to a c r u c i b l e . The c r u c i b l e

and c o n t e n t s w e r e d r i e d to a c o n s t a n t weight a t 1 0 0 ~ ~ The

. c o n t e n t was

then heated over a flame a t r e d h e a t f o r 2 0 minutes. The c r u c i b l e was

cooled i n d d e s l c d t o r m d weighed. The percentage of crude f i b r e was

c a l c u l a t e d ds follows:

Crude f i b r e (%) 5

w t , of sanple -
l o s s i n w t . x 100
1
I

3.7.4 ~ e t t : r m i n-a-t-i o-n-of A s t r

-

The A.O.A.C.(1980) method of a n a l y s i s was used f o r ash c o n t e n t

determinations.

About 5g of t h e stunple was weighed and heated i n a 50ml p e t r i d i s h

a t 1 0 0 u~n ~t i l water was expelled. Few drops of pure o l i v e o i l w a s added

and t h e mixture heated over flame u n t i l swelling stopped. The d i s h was

then placed on a furnace a t 525Oc and l e f t t h e r e u n t i l white ash was

obtained. ,The a s h was moistened.with water, d r i e d on a steam b a t h and

t h e n on a h o t p l a t e , t h e r e a f t e r re-ashed a t 5 2 5 ' ~ t o c o n s t a n t weight.

The a s h c o n t e n t w a s c a l c u l a t e d a s a percentage of t h e o r i g i n a l

weight
. . of t h e sample as follows:

Ash I w t . of ash x 100

wt. of sarnple

3-7.5 T o t a l Carbohydrate determination:

0,2g of ground sample w a s mixed w i t h 50ml of d i s t i l l e d water i n a

b o i l i n g beaker. .. .
3ml of concentrated I l c l . w d s added and t h e mixture b o i l e d

u n t i l complete hydrolysis. The c o n t e n t of t h e f l a s k was cooled &:d

i
n e u t r d l i s e d with 5N NaO1.I s o l u t i o n . The hydrolysate was then t r a n s f e r r e d

i n t o a 100rnl volumetric f l a s k and the volume rnade up t o 100ml w i t h

d i s t i l l e d water. 0.2ml of this s o l u t i o n w a s p i p e t t e d and mdde up t o 2ml

(10 f o l d d i l u t i o n ) w i t h water f o r the dctemir?at,ion.

G ~ U C O Swas
~ determined using t h e anthrone r e a g e n t (Deriaz, 1961). A

stock glucose s o l u t i o n of O.8mg/ml was prepared. Glucose s t a n d a r d s were

prepared by ddding O m l , 5m1, 10m1, 15m1, 20ml, and 25ml of t h e stock

s o l u t i o n i n t o s i x d i f f e r e n t l O O m l volumetric f l a s k s . The volumes were

made ui) t o the mark with d i s t i l l e d w d t e r .

l m l of each of the s t a n d a r d s o l u t i o n s and test simples were

r e s p e c t i v e l y p i p e t t e d i n t o test tubes. To each of t h e test t u b e s , 5ml

of dnthrone r e g e n t wcjs added and p r o p e r l y mi,xed. These were covered ond

inunedidtely p u t i n a b o i l i n g water b a t h f o r 20 minutes f o r t h e c o l o u r to

develop. They were cooled and t h e i r absorbance measured a t 620nm. From

t h e s t a n d a r d glucose c a l i b r a t i o n curve p l o t t e d , t h e c o n c e n t r a t i o n of

glucose i n t h e t e s t sample was r e a d off.

Calculdtion:

% total. carcohydrate P mg of glucose e q u i v a l e n t t o sample absorbance

from graph x conversion f a c k o r ( 2 5 ) x d i l u t i o n f a c t o r ( 1 0 ) .

3.7.6 G e l e t i n i z a t i o n temperature determination:

Ttm g e l e t i n i z a t i o n temperature determination was c a r r i e d o u t in

accordance w i t h t h e rnethod of Novellie ilnd ~ c & t e :(1961).

 29 of ground sample w a s p u t i n 40ml of water i n a l a r g e t e s t - t u b e , and

quickly brought t o t h e desired t u n p e r a t u r e w i t h good s t i r r i n g f o r a t l e a s t

5 minutes heating. ~ f t e gr e l e t i n i z a t i o n , * e mixture was cooled t o 30°c

md placed i n a c o n s t a n t temperature water bath. Malt e x t r a c t (40rnl) was

added and a f t e r a thorough i n i t i a l mixing, t h e mixture was s t i r r e d h a l f

'
hourly f o r 3h. A s m p l e 20ml was withdrawn and added t o 30ml of 0,SN

NaOH s o l u t i o n i n a 250ml ' v o l u m e t r i ~f l a s k . The mixture was made t o volune

asld t h e s u g a c o n t e n t detennincd by Benedict's q u a n t i t a t i v e method(Plummer,

1977). For t h e blank, equal weight of sample was p u t i n t o 40ml of

w d t e r and e x t r a c t (40ml) added to t h e u n g e l a t i n i z e d s t a r c h a t 30°c f o r 3h,

T h e reducing sugar w d s s i m i l a r l y determined a f t e r stopping t h e enzyme a c t i o n

with 30ml of 0.5N NaOH s o l u t i o n .

25ml of ~ e n e d i c t l sq u a n t i t a t i v e r e a g e n t was measured i n t o a l O O m l

conical flask. To t h i s was added 3g of anhydrous Na--CU3 and a few p i e c e s

of anti-bumping granules. The mixture was heated over a bunsen flame and

when i t b o i l e d vigorousl.y, t h e s u g a r s o l u t i o n was run i n slowly from a

tiurt:tte. When ~i bulky white p r e c i p i t d t e w d s formed, t h e s u g a s o l u t i o n was run

i n more slowly u n t i l t h e l a s t t r a c e s of b l u e had disappeared. The t i k e

which was t h e volume of s u g a r r e q u i r e d was noted.

The c o n c e n t r d t i o n of t h e s u g a r was c d l c u l a t e d from t h e following

f d c t o r s ; 25ml of Benedictcs r e a g e n t i s e q u i v a l e n t t o 50mg of glucose, 53mg

8

of f r u c t o s e , 68.8mg of l a c t o s e , 74 mg o f maltose o r 49mg of hydrolysed

sucrose.
 m g s u q a per 100ml

where f P. sugar f d c t o r

3 7.7 s t a r c h deterinination i n s t a r c h c o n c e n t r a t e by h y d r o l y t i c method:

A t y p i c a l method employing a c i d h y d r o l y s i s f o r t h e determination of

sterrch i n f-lours i s t h a t of Radley(1976) employed i n t h i s s t u d y as follows:

2-59 of d r y s t a r c h c o n c e n t r a t e w a s mixed w i t h 50ml o f c o l d water and

dllowed t o stand f o r lh. of H c l and 150ml of water were added t o the

2011-11

suspension. his w a s p u t i n t o a round bottomed f l a s k and r e f l u e d f o r 2h.

The c o n t e n t of the f l a s k was cooled and n e u t r a l i s e d w i t h 5N NaOH. The volume j

I
1
w a s l a t e r mdde up to 2SOml. lml was p i p e t t e d into a 100ml volumetric f l a s k

m d t h e volume made up t o t h e mark w i t h d i s t i l l e d wat

t;Jr . Glucose was

determined using t h e anthrone reagent(Deria2, 1961). A series of glucose

s o l u t i o n s were prepiired, s o t h a t l m l c o n t d n s 0.04 - 0.2mg, and.these

I
were used t o c a l i b r a t e t h e glucose s t a n d a r d curve. .. ,

I m l of each of the s t a n d a r d s o l u t i o n and t h e test sample was r e s p e c t i v e l y

p i p e t t r d i n t o test- t*&es. To each, 5ml o f anthrone r e a g e n t was added and

I
!
1
' - --r- * - y mixed.
-.7,7nC.."l
These were covered and immediately p u t i n a b o i l i n g water

b a t h f o r 20 minutes f o r t h e c o l o u r t o develop. They were cooled and t h e i r

absorbance measured a t 6 2 0 m a g a i n s t a blank which contained only I m l o f ,

water and 5ml of anthrone reagent. The concentcation of t h e test sample

. . .
wcls obtained f r a n t h e & s o r b a x e by e x t r i r p o i a i i a ~ . The mass of glucose d

wclv o b t d n e d by c d c u l d t i o n s involving t h e c o n c e n t r a t i o n s and d i l u t i o n s

mdde. The mclss of s t a r c h was con:eq!ently obtained from t h e mass of

j l u c o s c using t h e r e l a t i o q

~ , i s sof glucose x 0.9 = Mass o f s t a r c h .

production of Malt based Syrups:

3 , ~ .1 *wort p r e p a r a t i o n by t h r e e s t a g e decoction Mashing Method from

Sorghum malt.

The method, c u r r e n t l y adopted f o r masl~ingb a r l e y m a l t a s described

-
by ~ o u g he t , &.,(1981) was used i n t h i s work as follows:

S i x t y grams of Sorghum malt m i l l e d t o 1-2mm p a r t i c l e s i z e ( i n a Thomas

illy ill Model'ED 5 ) was mixed w i t h 32Gml of t a p water, t o g i v e 18.4%

mash. T h i s !as held a t 40°c f o r 30 minutes. One t h i r d p o r t i o n o f mash

was withdrawn, b o i l e d f o r 5 minutes, and qeturned t o the main mash. The

temperature of t h e mash r o s e t o SOOC and was maintained a t t h i s temperature

f o r 15 minutes a t a pH 6.5. (This was adjusted w i t h 2~ Ca(OHI2 ~ 0 1 ~ t i 0 n ) ~

One t h i r d p o r t i o n of t h e mash was again removed, b o i l e d f o r 5 minutes and

r e t u r n e d t o t h e main mash. The temperature of t h e mash was r a i s e d t o 60°c,

and t h i s was maintained a t a temperature range of 60-65O~ f o r 30 minutes. .

A f u r t h e r one t h i r d p o r t i o n of the mash was removed, b o i l e d f o r

5 .minutes and r e t u r n e d to t h e main mash. The temperature r o s e , and was

maintained a t 70-75O~ ( t h e m;shing o f f temperature) f o r 30 minutes.

3 w 8 2 E f f e c t s of varying mash c o n c e n t r a t i o n s and S x c h a r i f i c a t i o n p e r i o d s

on reducing s u g a r c o n t e n t s of worts in a t h r e e s t a g e decoction
mashing.

I n t h e mashing method d i s c u s s e d above, p r o t e o l y s i s was dllowed a t

4uDc and 50°c. s a c c h a r i f i c a t i o n by t h e malt amylases was encouraged by

holding t h e mash a t 60 - 6 5 O ~rvld 70 - 7s0c temperatures r e s p e c t i v e l y . Mash

c o n c e n t r a t i o n s o f 25%, 35%, and 45%, and t o t a l s a c c h a r i f i c a t i o n p e r i o d s of

I h , 2h, and 3h were employed i n t h e production of warts. The r e s u l t i n g '

reducing s u g d c o n t e n t s ( a s g l u c o s e ) determined a s d e s c r i b e d under s e c t i o n

"
3.9.4 .

3-8-3 E f f e c t of v x y i n g concentr~tt.jonsof -a l u c o m y l a s e
- ,
and uaccharif i c a t i o n
periods on t h e reducing sugar c o n t e n t s of wort.

Wort W ~ Yp r e p d e d from a 25% mash. Its pH was a d j u s t e d t o 4.3 w i t h a

0
3~ ~ c slo l u t i o n , and temperature maintained a t 55 C. Varying c o n c e n t r a t i o n s

of g l u c o a m y l ~ s c ; 0.09%, 0.10X and 0,15X (dry weight b a s i s of t h e mash) were

ddded r e s p e c t i v e l y , then s a c c h a r i f i c a t i o n c a r r i e d o u t f o r 12h, 24h, and

The r e s u l t i n g hydrolysates were n e u t r a l i s e d , f i l t e r e d and m a l y s e d f o r

reducing sugar c o n t e n t ( as g l u c o s e ) , as d e s c r i b e d under s e c t i o n 3.9.4 '

3 . 8 -4 Ma1t bdsed- Syrup production:

50y o f sorghum m a l t m i l l e d t o 1-2mm p a r t i c l e s i z e ( i n a Thomas wiley

M i l l , Model ED-5) was mixed w i t h 2 0 h l o f tap water t o g i v e 25% mash. The

ptf of t h e r e s u l t i n g wort w a s a d j u s t e d t o 4.3. X t r temperature was maintained

a t 5 5 ' ~ using a t h e n n o s t a t i c a l l y c o n t r o l l e d water bath. 0.15%- glucoamylase

(D.W.B of t h e mash) was added and incubated f o r 24-72h (with constant

shaking) u n t i l t h e d e s i r e d DE v a l u e was a t t a i n e d .

The m a l t h y d r o l y s a t e s o l u t i o n produced was n e u t r a l i s e d w i t h 2M Na2C03

solution. A t e n f o l d d i l u t i o n of it was made, f i l t e r e d ( w i t h f i l t e r paper)

and. f i n a l l y concentrated to a syrupy c o n s i s t e n c y by evaporation on a

b o i l i n g water bath.
 / ~ h r e es t a g e decoction mashinq/
1
J
/~nzyme s a c c h a r i f i c a t i o n l
1

. .
JI
h i n i s h e d product ( ~ a l syrup)/
t

~ i g .5. Flow c h a r t of m a l t based syrup production. . \

Glucose Symp Production:

3.9
The acid-enzyme(dua1-stage) conversion method w a s used i n t h e glucose

syrup production. I n t h e f i r s t s t a g e , which w a s a c i d l i q u e f a c t i o n , s t a r c h

w d s mixed w i t h water t o form a suspension of s i u r r y , conicaininy 25% d r y

starch. his was poured i n t o a pyrex f l a s k and i t s pH a d j u s t e d to 1.8

w i t h 3M H c l s o l u t i o n , The f l a s k and its c o n t e n t w a s autoclaved f o r 20 minutes,

The r e s u l t i n g h y d r o l y s a t e w a s cooled, and n e u t r a l i s e d t o pH 6.0. This

was l a t e r f i l t e r e d i n t o a flask, and t h e second s t a g e of the conversion,

enzyme s a c c h a r i f l c a t i o n e f f e c t e d by a d d i t i o n of 0.15% g l u c o a m y l a s e ( 9 ~o~f

s t a r c h ) , and holding i t i n a w a t e r b a t h ( w i t h c o n s t a n t a g i t a t i o n ) a t
was determined by t h e d e s i r e d DL l e v e l . The r e s u l t i n g s t a r c h h y d r o l y k i t e

was concentrated by e v a p o r a t i o n on a b o i l i n g . w a t e r bath,

hon - exchange
4
deionized/
I

Fig. 4. Flow c h a r t of Acid-enzyme converted glucose syrup production.

3.9 Determination o f some P r o p e r t i e s of Syrups:

3.9.2 Determination of S p e c i f i c q r a v i t y / ~ e g r e ebaume o f the Syrups.

The s p e c i f i c g r a v i t y of t h e syrups were determined according t o t h e

procedure d e s c r i b e d in ~.O,A.C.(1980) method of a n a l y s i s .

 A 50ml s p e c i f i c g r a v i t y b o t t l e was f i l l e d w i t h d i s t i l l e d water,
0
stoppered irnd immersed i n water b a t h a t 20 C. A f t e r 30 minutes-, t h e

s p e c i f i c g r a v i t y b o t t l e was removed, d r i e d with f i l t e r paper and weighed.

The s m e procedure was repaated w i t h t h e s m p l e s . The s p e c i f i c g r a v i t y

of t h e samples were c a l c u l d t e d as follows:

SG P weight of l i q u i d held i n SG b o t t l e
weight of water held i n SG b o t t l e .

Uegree baurne ( ' Me' 1

T h i s i s r e l a t e d t o S p e c i f i c g r d v i t y b y , t h e following fonnula(Corn

r e f i n e r s Assacidtion, 1 9 6 5 ) .

3 ,go 3 " Determkidtion o f percentage t o t a l s o l i d s of t h e Syrups.

A sample of t h e syrup w a s t r a n s f e r r e d i n t o a p r e v i o u s l y d r i e d and

weighed s t a i n l e s s steel dish. The d i s h and c o n t e n t was l a t e r placed on a

boiling water b a t h and evaporated to dryness, This was weighed and then

placed i n .an oven and d r i e d f o r 3h a t 1 0 5 ~ ~The

. d i s h was r e t u r n e d t o t h e

oven and weight checked a t thirty minute i n t e r v a l s u n t i l no f u r t h e r l o s s

i n weight could be detected. The d i s h was cooled i n a d e s i c a t o r f o r

20 minutes and t h e weight taken.

ti'
The percentage t o t a l s o l i d s , was c a l c u l a t e d thus:- .

% t o t a l s o l i d s = W t of sample a f t = d r y i n g x 100
W t o f sample bekore 'drying

! ' 3.9.4 J Percentage reducing s u g a r conten t / ~ e x t r o s ee q u i v a l e n t ( DE.) value

determination.

Reducing sugar c o n t e n t (as glucose)/Dextrose eqUivalent(DE) values of

thesyrup s m p l e s were detenninfid &cording t o t h e I n s t i t u t e of Brewery

I.O.U.(1977) method of analysis as follows:

It% w/v s o l u t i o n was ,prepared by weighing.25,OOg of t h e syrup i n a

. . .

g l d s s d i s h and d i s s o l v i n g same b y . g r a d u a 1 s t i r r i n g i n warm water. his

w d s t r a s f e r r e d q u a n t i t a t i v e l y to a 2501111 graduated f l a s k , and a f t e r
0
a d j u s t i n g t h e temperature t o 20 C, it was mdde up t o t h e mark a t t h a t

temperature. 251111 of t h i s s o l u t i o n was p i p e t t e d i n t o a 250ml graduated

f l a s k and d i l u t e d t o mark a t .20°c. T h i s was mixed w e l l and f i l t e r e d . It

wris used a s t h e " d i l u t e d solution".

25ml of mixed Fehlings' s o l u t i o n was pipetted i n t o a 250rnl S a i l i n g

f l a s k ; and m almost sufficient o f t h e d i l u t e d s o l u t i o n was added from t h e

b u r e t t e t o t h e cold f e h l i n g s ' s o l u t i o n t o e f f e c t r e d u c t i o n , s o t h a t , i f

p o s s i b l e , n o t more than l m l was r e q u i r e d l a t e r t o complete t h e t i t r a t i o n .

The c o n t e n t s of t h e f l a s k was mixed, and heated over a w i r e gauze, k e p t i n

moderate e b u l l i t i o n f o r 2 minutes and t h r e e drops of methylene b l u e i n d i c a t o r

added without removal of t h e flame, then t h e t i t r a t i o n completed i n one

minute w i t h continuous e b u l l i t i o n .
I
The end p o i n t ( d e c o l o r i z a t i o n of t h e methylene b l u e ) was taken a s t h e

volume 'dt which t h e r e a c t i o n mixture turned red. The t i t r e was recorded.

Calculations:

The r e s u l t s were,ccilculated a s t h e most a p p r o p r i a t e sugar (glucose o r

maltose) using t h e a p p r o p r i a t e f a c t o r from t h e l a n e & Eynon t a b l e . For t h e

d i l u t i o n given, t h e percentage reducing s u g a r i n t h e sample ' a s i s g

% ~ e d k i ns u~g a r = Lane & Eynon f a c t o r x 100

Titre.
 Dextrose Z q u i v a l e n t ( ~ ~r )% reduclnq sugar ( a s glucose) x 100
% total solids

3 m9.5 . - Deterrnl-nation o f c o l o u r of t h e syrups:

The c o l o u r of the syrups were determined by t h e procedure d e s c r i b e d

i n A.O.A.C(l980) method of a p d y s i s as follows:-

59 c e l - i t e was added t o 100rnl f i l t e r e d syrup, mixed and f i l t e r e d

through f i l t e r pclper. The f i r s t 40ml f i l t r a t e was r e t u r n e d t o t h e f i l t e r .

Absorbmce of c l e a r t o t a l f i l t r a t e was determined a t 430m using a

spectrophotorner. Syrup c o l o u r was c a l c u l a t e d a s . follows:

colour = 1 0 . x x correction t o 4' cell s i z e . c ~ l o u r was r e p o r t e d t o t h e

n e a r e s t 0.05 u n i t s .
 CHAPTER 4

4. RESULTS AND DISCUSSIONS

4.1 ~ a l t i n gc h a r a c t e r i s t i c s o f t h e c e r e a l p;rains, sorghum and millet:

Table i shows t h e malting c h a r a c t e r i s t i c s of t h e c e r e a l g r a i n s

used. The v a l u e s of 33.30g and 6.8g 1000-kernel weight obtained r e s p e c t i v e l y

f o r sorghum and millet g r a i n s were s m a l l e r compared t o 50.9g f o r b u l e y

reported by s i n g h and ~ a u r o ( 1 9 7 7 ) . However, s i n c e t h e s e g r a i n s are

s m a l l e r i n s2ze, it is expected t h a t t h e weight would be smaller. his

agrees w i t h t h e suggestion made by Nout and Davis(1982) t h a t i n l a r g e

s c a l e malting, modification i n p l a n t cm be e f f e c t e d t o t a k e care of t h e

d i f f e r e n c e s i n s i z e of t h e g r a i n s .

The percentage f o r e i g n s e e d s and broken k e r n e l s gave 0.32% and 0.56%

re3;)ectively f o r Sorghum and m i l l e t g r a i n s . These r e s u l t s show t h a t

broken k e r n e l s and f o r e i g n s e e d s are r e l a t i v e l y low from t h e bulk g r a i n

samples.

tiough et &., ( 1981) r e p o r t e d t h a t f o r good malting q u a l i t y of b a r l e y ,

minimum germinative energy (G.E) r e q u i r e d i s 96%. However, i n t h i s study,

G.E. of 82% and 76% were obtained r e s p e c t i v e l y f o r sorghum and ' m i l l e t

grains. Aniche(l302) obtained 60% C.E. and 98.5% germinative ' c a p a c i t y

(G.C. ) for L 181 sorghum v a r i e t y ; while Singh and Tauro(1977) r e p o r t e d

7% germinative energy f o r millet. G e r m h a t i v e c a p a c i t y (G.C.) o f 90%

and 85% were obtained f o r sorghum and m i l l e t r e s p e c t i v e l y i n t h i s study,

iind s i n c e G.C i s a measure of percentage of l i v i n g c o r n s under aided

c o n d i t i o n s , i t i s t h e r e f o r e suggested t h a t hydrogen peroxide be used

 62.

durlriy 111dlthg of sorghum iuld m i l l e t g r a i n s t o enhance germindtivt: cclpacity.

MAIJTING CKARACTERISTICS OF l l i E CERAL GRAINS: SORGHUM AND MILLET

~haracteristics Sorghum Millet

1000 - kernel weight(g) 33.30 6 .8

% Foreign seeds and broken k e r n e l s ' 0.32 0.56

Germinative energy (% ) 82 76

O p t h u m m j l t i n g c o n d i t i o n s of t h e Cereal grains: Sorghum and Millet:

Figures 6 t o 10 p r e s e n t t h e optimumgmalting c o n d i t i o n s of t h e

c e r e d l grains. The v a r i a t i o n o f moisture c o n t e n t ( % ) a g a i n s t s t e e p time(hours1

i.s shown i n f i g u r e 6 , The r e s u l t i n d i c a t e a sharp rise i n water uptake

during t h e f i r s t 1'0 hours. F u r t h e r s t e e p i n g above 20 hours showed marginal

increases. I t i s e v i d e n t from t h e r e s u l t s t h a t sorghum g r a i n s absorbed

moisture fclster than m i l l e t g r a i n s , t h i s may be due t o i t s r e l a t i v e l a r g e

corn s i z e . -
Dahlstron e t g . , ( 1 9 6 3 ) observed t h a t l a r g e r c o r n s absorb water -
more r a p i d l y than s m a l l e r ones i n i t i a l l y , and t h e d i f f e r e n c e i n water

absorption a f t e r 24 hours is marginalised. Also Hartong and Kretschmer

(1961) found t h a t samples of g r a i n s t h a t absorbed w a t e r f a s t e r gave b e t t e r

m a l t s than g r a i n s t h a t absorb water more slowly.

0
The r e s u l t obtained from t h e p l o t of d i a s t a t i c power( L) a g a i n s t

steep t i m e ( h o u r s ) f i g u r e ' 3 , showed t h a t t h e o p t h u m s t e e p t i m e Was

 1 1 I 1 1 I I
\ I
10 20 30 40 50 . 60 70 80
Steeplng t i me (hours)
Fig. 6 : Moisture content (%) against s t e e p i n g t i m e ( h o u r s ) .
Sorghum grain
i Millet grain
 20 ' 30
St seping
Fig. 7 : Diastotic power (%) against st;esping tima (hours) a f t e r
4 days o.f g e r m i n a t i o n . - ..
b-d 8 Sorghum, o-o 8 Millet '
50 kwurs. A t t h i s time, the d i a s t a t i c powers of the two c e r e a l s were

mdx.i.mum a f t e r the grains w e r e allowed t o germinate f o r four days. Furthermore,

extrapolating t h e 50 hours optimum s t e e p t h e t o r e s u l t s of the p l o t of

moisture content(%) against time(hours) f i g u r e fj, gave variously 38%

and 33% optimum moisture contents f o r sorghum and m i l l e t grains respectively.

I

out and Davis (1982) obtained 45% moisture c o n b n t a f t e r 35 hours and

20 hours of steeping sorghum QAndivo' and sorghum 'igumba, respectively.

'
The optimum gemination period of 5 days was obtained after steeping
I
the g r a i n s f o r 50 hours. This r e s u l t was deducted from t h e p l o t of

d i a s t a t i c power (OL) against germination periods(days), f i g u r e 8 The

'
0
r e s u l t a l s o i n d i c a t e t h a t rnJxknum d i a s t a t i c pX!=rs of 3 2 O ~and 2 7 L were

obtained respectively f o r sarghum and m i l l e t malts under the same s t i p u l a t e d

, I

malting conditions. The r e s u l t shows t h a t t h e r e was no d i a s t a t i c power I

measurable in ungerminated grain b u t rises sharply fran 1-3 days reaching

i t s peak a f t e r 5 days of gemination. This suggests t h a t d i a s t a t i c enzymes

absent i n ungerminated g r a i n develops with germination. I t has been established

t h a t alpha anylase was not present i n the ungerminated sorghum grain and t h a t

the a c t i v i t y was only measurable a f t e r germination(Aisien et z., 1983). i

I

Figure 9 -shows the r e s u l t of malting l o s s (%I measurements during

the various periods of germination(days). The r e s u l t showed t h a t the

malting l o s s of g r a i n s increased with increase i n the period of germinatim.

S i g n i f i c a n t increases i n malting l o s s were recorded between 2-4 days of

germination, which correspond t

a t h e periods of s i g n i f i c a n t drops in

1000 - kernel weights during gemination. The ranges of 12 - 16% and 16.20%
 Germination period (day,)
Fig. 8 : Diastolic power (%I a g a i n s t germination periods(days)
a f t e r 50 h o u r s of steeping. I

H'= Sorghum, M = Mill8 t

I 1 I 1 1 1
2 3 4 5 6 7
Germinat i o n p k r i o d ( doyo )
9 : Molting loss (%I against perminotion period ( d a y s ) .
04a Millet molt
M Sorghum ma I t
malting l o s s e s were obtained respectively f o r sorghum and m i l l e t m a l t s

covering 4 to 7 days of gemination. The malting l o s s f o r barley have

been given by Hough et: ( 1981) ae 6~12%. The high malting l o s s of

t h e millet grains could be due t o excessive aeration during sixaping

leading t o grains growing uncontrollably dullng germination according to

Houghe, &., (1981). Malting l o s s could also r e s u l t from long steeping

period a s materials trend to be leached i n t o t h e s t e e p water. .To reduce

malting l o s s , it,i s suggested t h a t the f i l t e r paper8 used should n o t be

s a t u r a t e d with water, o r I n the a l t e r n a t i v e , malting f l o o r s should be wed.

Relault of kilning s t u d i e s , ( f i g u r e 10 ) shows t h a t moisture r e l e a s e is . ;

reciprocally r e l a t e d to moisture absorption. S i g n i f i c a n t l o s s i n moisture

w a s recorded after 12 hours of k i l n i n g a t 4s0c. With regards to moisture

l e v e l i n r e l a t i o n to storage q u a l i t i e s of malta, optimum k i l n i n g can be

c a r r i e d o u t a t 4 5 O ~f o r 24-48 hours depending ~n the moist- l e v e l of

t h e green malt and the moisture content of the m a l t required. The e f f e c t

of temperature on the m a l t characters was n o t investigated, however,

other workers; Nout and Davis ( 1 9 8 2 ) and Novellie,(1962) showed t h a t only

kilning a t .70°c r e s u l t e d in a significant loss i n diastatic activity,

kilning in the range of 4 0 - 6 0 ~caused

~ only negligible destruction.

4.3 Evaluation of ma1t;l'squality characteristics:

The malts* q u a l i t y c h a r a c t e r i s t i c s were evaluated and the results

presented on t a b l e 5 .. Generally, low Cold Water Extract(C.U.E), high

Hot Water ~ x t r a c t ( ~ . W . ~ . and

) la static P o w ~ ( D P )are
. i n d i c a t i v e of good

m a l t characteristics. Hough et; %,(1981) reported ~ a l u e of

s 307 ~ O h g
 Kilning period (hours)
ig. 1 0 : Moisture content (%) agoins* kilning p e r i o d ( hours) at 45OC.

o-a 8 M i l l e t malt, - k + - ? 8 $ o t r g h u m m a l t ,
(HUE , 18.6% (CWE) and 6 3 ' ~(DP) f o r barley malt!. Tha corresponding values

obtained i n this atudy f o r sorghum and m i l l e t malts( t @ l e

I
f, .' were
r e l a t i v e l y inadequate to pass f o r good malts. The high C.W.E obtained

may be due t o malting a t high temperature of around 2 8 ' ~ ~

aa against t h a t
0
f o r barley which i s usually a t 15 C o r less (Preece, 1954). as w e l l ao a

higher moisture content achieved by sprinkling of water. Improvements I n

the malting conditions f o r t h e grains could considerably reduce the high

C-9-E-, and increase the H.W.E. and D.P. values obtained. However, the

values obtained f o r Sorghum malt i n ' t h e three,q u a l i t y c h a r a c t e r i s t i c s arc

comparatively more encouraging than those of m i l l e t malt.

THE MALT'S C\UALITY CHARACTERISTICS

Characteristics Sorghum m a l t M i l l e t malt

Cold Water Extract, CWE(%)

~ o water
t e x t r a c t , HWE ( ~ O / k g )

4.A Determination .
of Optimum pH and Temperature conditions f o r malt's
- .
amylase a c t i v i t y .

The r e s u l t s of t h e optimum conditions (pH and tempersture) f o r m y l a s e

a c t i v i t y of t h e malts were as pretsentsd i n f i g u r e s 12 ' and -13 respectively.

Figure 12 shows t h a t a t optimum pH range of 6-7, t h e amylase a c t i v i t i = e

I

of both sorghum and m i l l e t malts were maximum. Also f i g u r e 13 reveals

t h a t amylase a c t i v i t y of sorghun m a l t peaked a t optimum temperature range

 - mg Maltose in 3 m l solution
Fig . I 1 : .Maltose calibration c u r v e . '
 40 50 60 70 80 90
Temperature ( O C )

.IFi g 8 : Optimum ternperoture determination far ornylo~eactivity

in both millet, ond sotghum m a l t $. I tb
o f 60-7o0cI while t h a t of m i l l e t malt b e c q e m a x i m u m a t 40-50°c. The

optimum p~ value r m g e of 6-7 obtained i n t h i s etudy agrees with thQ

values reported by ahembe &-,(1988). S i l l s and Stewart(l984) reported

optimum pH range of 5.5-6.5 f o r p u r i f i e d b a r l e y alpha amylase. ~ough

-
et &.,(1981) p o i n t s out t h a t pH value of 6.0 is q u i t e s u i t a b l e f o r
0
anylase a c t i v i t y a t 65 C. The r e s u l t i s a l s o in agreement with t h e pH

optima of 5.2 (Preeoe, 19541, f o r /3-amylase, and 7.O(Worthington, 1979)

f o r alpha amylase enzymes i n barley.

shembe 6 &.,(1988) reported 35-45O~ temperature optima f o r m i l l e t

and 50-75Oc $or sorghum m a l t s @ amylase a c t i v i t i e s . Also S i l l s and Stewarts

(1984) reported values of 3 5 - 4 0 ~f ~o r p u r i f i e d b a r l e y alpha amylase.

A

Chandraskhara and swminikhan ( 1958) showed optimum d P e r a t u r e f o r both

alpha and b-anylases in p e a r l m i l l e t t o be 60°c. The &mporature optima i n

both cases a r e below g e l a t i n i z a t i o n temperature ranges of millet(70-80°c)

and sorghum (68-80°c) obtained i n t h i s study; a necessary p r e r e q u i s i t e f o r

e f f e c t i v e degradation of s t a r c h by amylases. T h general poor malting

q u a l i t y of sorghum and m i l l e t ! g r a i n s as a l s o reported by o t h e r authors is

thus p a r t l y due to this f a c t .

4.5 ~ r o x i m & / ~ h e m i c a ~analyses

. of t h e sorghum/millet g r a i n s and maltst

The r e s u l t s of proximate analyses (moisture content, crude p r o t e i n ,

f a t , crude f i b r e , and Ash) t o t a l carbohydrate. and ' g e la t l n i zation temperature

ranges of the g r a i n s and m a l t s used i n producing the syrup8 a r e

in.table 6 , , he proximata c m p o s i t i o n values obtained f o r m i l l e t g r a i n s

and i t s malt w e r e c l o s e to those reported by Opdrtu et . , 1 9 8 1 Also

Aniche (1982) proximate composition values reported f o r Sorghum malts

(table 2 ) caipclred f avourably with the values obtained i n t h i s work.

The Sorghum values f o r protein, 10.40%, c l o s e l y agree with 10.9% reported

by Hough et . , 1 9 8 1 . The r e s u l t show t h a t sorghum has lower values

of f a t , and higher values of carbohydrate and p r o t e i n than m i l l e t . Grains

used f o r syrup production should be low i n fat and p r o t e i n b u t r i c h i n

t o t d l carbohydrate e s p e c i a l l y s t a r c h . Cereals with higher f a u p r o t e i n

contents are considered unsuitable f o r glucose syrup manufacture s i n c e

they have detrimental e f f e c t s on f l a v o u r ar\d c o n t r i b u t e t o colour v i a maillard

reaction. I n malt based syrup production, where colour i s r a t h e r required,

the high p r o t e i n content f a c t o r i s no more necessary, t h e r e f o r e sorghum

g r a i n is b e s t s u i t e d f o r t h e malt syrup production.

TARLE 6.

CHEMICAL/PROXIMATE ANALYSES OF SORGHUM/MLLLET GRAINS AND MALTS

~ i l l e t Sorghum
Grain Malt Grain Malt

Moisture content(%) 9-10 ' 6.50 10.12 7.50

Crude p r o t e i n (%I 8.30 10.50 10.40 11.30
4.6 Chemical analyses of s t a r c h e s extracted from m i l l e t and sorghum, grains:

Starches were e x t r a c t e d from m i l l e t and sorghum g r a i n s and subsequently

subjected t o chemical analyses. The r e s u l t s obtained are presented on

t 5 1 e 7. The lower s t a r c h contents of 76*40(millet) and 83.40(sorghum)

r e l a t i v e to values obtained by Brenner e t -- d-.-.P.-..

&.,\LYQO~ and nuisr &, &.,
(1980) may be due t o the unsophisticated equipment which was used during

the s t a r c h e x t r a c t i o n process. The p r i n c i p a l parameter i n t h e r e f i n e d

s t a r c h concentrate which a f f e c t s both t h e q u a l i t y and volume of glucose

syrup i s t h e s t a r c h content. The higher the s t a r c h content, the g r e a t e r

t h e reducing sugars obtained on hydrolysis., The comparatively higher p r o t e i n

values of 2.1% ( m i l l e t s t a r c h ) and 2.4%(sorghum s t a r c h ) a s a g a i n s t 0.2016

reported by Hulse & &.,(1980) may equally be due t o inadequate separation

i

of gluten from s t a r c h . The l e v e l s of f a t 0.83% ( m i l l e t s t a r c h ) and 0.66%

(sorghum s t a r c h ) are a l s o higher than 0.003% reported by B r e n n e r c 2..

(1968). Therefore, a more sophisticaited equipment than tk one used i n

t h i s work i s suggested f o r s t a r c h e x t r a c t i o n i f glucose syrup with a near

water-white colour i s desired.

Table 7 shows t h a t the g e l a t i n i z a t i o n temperature ranges of t h e

extracted' refined s t a r c h e s are lower than t h a t of the corresponding g r a i n s

(table 6) thereby rendering t h e r e f i n e d s t a r c h more s u s c e p t i b l e t o

amylalytic degradation. However, sorghum s t a r c h , having a lower g e l a t i n i z a t i o n

temperature r a n g e , l w e r f a t and ash contents than m i l l e t s t a r c h is preferred

i n glucose syrup production.

 TAJ3LE. 7.

CtiEMlCAL ANALYSES OF STARCIIES FROM MILLET AND SORGHUM GRAINS

Millet s t a r c h Sorghum s t a r c h

~ o iture
s content (%I 9.2 8.6
b ;'
Crude p r o t e i n (76) ' 2.1 2 .4

Fat (%)

Crude f i b r e (%I 0.71 0.27

Ash (%I 0.60 0.40

Starch content (%) 76.40 83.40

4.7 E f f e c t s of varying mash, glucoamylase concentrations and s a c c h a r i f i c c ~ t i o n

p e r i o d s on reducing sugar contents of t h e wort syrup:
-
Figures 15 and 16 show r e s p e c t i v e l y t h e r e s u l t s of t h e e f f e c t s of

varying mash concentrations and time; glucoamylase concentrations and time;

on t h e reducing sugar content of mort syrup. Results show t h a t makimum

conversion was a t t a i n e d a t t h e lowest s u b s t r a t e concentration, 25% mash;

highest enzyme-substrate r a t i o , 0.15% glucoamylase and a t a longer

s a c c h a r i f i c a t i o n time. However, t h e r e are f e a s i b l e ranges of i n t e r a c t i n g

v a r i a b l e s during the s a c c h a r i f i c a t i o n process of syrup production. The

25-35s s u b s t r a t e concentra$lon range s t r u c k a b a l e e b e t w e e n t h e c o s t of

removing excess water from hydrolysates a g a i n s t higher conversfon t o glucose

a t t a i n a b l e a t low s u b s t r a t e concentration, Also, 0.10 - O,fi.% enzyme-

s u b s t r a t e r a t i o i s reasonable s i n c e c o s t of enzyme balanced a g a i n s t

s a c c h a r i f i c a t i o n and equipment required f o r long s a c c h a r i f i c a t i o n . The

 0 ~08 0.12 0.16
Glucose conc. ( m q / m l )
Fig.14 Glucose standard curve. .. .
 T i m e <hours) .
Fig . I5 Effect o f varying mash concenirot.ion and time on
reducing sugar .;content of the worta

= 4 5 % mash, H= 3S0/, mash , )++( 25'10 mash

 500-

480-

460-

% 440-
0
0
L
Q,

a 420-
Q)
Ln
2
C
M

g400- ,
u'
E

380-

360 J

36
0 12 Ti me 24
( hours
Fig. 16 : E f f e c t of w r y i n g concentrations of g l u c o a m y l a r e , . ~ n d
time on the reducing sugar content o f the wort syrup.

= 0 . 0 5 % of glucoarnylase
M = 0 10 % of giucoamylase
w, = 0.15 O k of glucoamylase
s a c c h a r i f i c a t i o n range of 12-72 hours chosen f o r the experiment w a s based

on enzyme-substrate r a t i o used and extent of corrverlion cle,aircd, The

optimum temperature range of glucoarnylase a c t i v i t y is S O - ~ ~ O C , where the

upper l i m i t i s dependent on glycoamylasestability, and lower l i m i t on neid

t o i n h i b i t microbial contamination during long saccherrification periods.

Furthennore, the pH 4-5-0 i s dependent on aource of glucoanylase, and a l s o

eug& are most s t a b l e a t this pH range.

4.8 Properties of malt based syrups and acid-enzyme converted glucose syrups: !
Tables 8 and g show the r e s u l t s of analyses i n some properties of
i
I
malt based syrups and glucose syrups produced i n this study. The r e s u l t s !
show, three ,
c l a s s e s of syrups; Regular ( 4 2 ~ ~intermediate
) (52~~1,

and High ( 6 9 ~ ~ j Conversion

. syrups were produced f o r malt and glucose

syrups respectively.

~ 42OF3ef density ware obtainad f o r m a l t syrups

The values of 4 3 ' ~and

and glucose syrups respectively, The higher value f o r malt syrup may be

due t o i t s r e l a t i v e l y higher content of cellulosfic materials.

The r e s u l t a l s o show t h a t t h e ash content of m a l t syrup was 0,4% while

that of glucose syrup was 0,65%. Junk and Pancoast,(l973) noted t h a t t h e

normal concentration range of ash i n corn syrup was from 0.1 - 0.3%. The

higher ash content of glucose syrup obtained may be as a r e s u l t of s o d i m

chloride, which i s derived primarily fran the n e u t r a l i z a t i o n of hydrochloric

acid w i t h sodium carbonate during the acid liquefaction process. '

 Colour values of 5.1 u n i t s and 2.2 u n i t s were obtained f o r m a l t ant!

glucose syrups respectively. Junk and Pancoast(l973) a l s o noted t h a t the

colour of newly refined'corn syr-p i s usually @water-whlte( and has a

value of 0.25 units. The higher value of malt syrup colour i s a t t r i b u t e d

t o the colour developed during the k i l n i n g process of malting. However,

the colour of the glucose syrup could be reduced through carbon &d ion-

exchang~r e f i n i n g processes.

he carbohydrate composition as glucose(%) and maltose(%) r e s u l t s

show a s i m i l a r trend on both tables. Generally, the values increase

proportionately across the three c l a s s e s of syrups, Also, mare g l u c o s ~

a r e i n each case produced than maltose, this i s expected since t h e

glucoamylase preparation from fermentations of Aspergillus niger contain

r e l a t i v e l y l e s s amounts of 8-amylase.

The lower glucose y i e l d i n m a l t syrup r e l a t i v e t o glucose syrup i n

t h e three c l a s s e s of syrups may be due to poor d i a s t a s i s of t h e m a l t s

0
amylases whose temperature optima, 60-70 C, is lower than the m a l t ' s

s t a r c h g e l a t i n i z a t i o n temperature range of 68-F30°c; Consequently, generating

8

more l i m i t branched dextrins which are less susceptible t o glucoamylase

than t h e l i n e a r dextrins(Abdul1ah A.,1963)

Results .also i n d i c a t e t h a t maltose contents of malt based syrups are

s l i g h t l y higher than thoge of glucose syrups{ this trend might be traced

to the r e l a t i v e . quantity of p-anylase which i s a major component of malt

amylases and a minor constituent of glucoamylase enzyme preparations from

-
Aspergillus niger.
 TABLE. 8.
PROPERTIES OF MALT BASED SYRUPS

Property Regular Intermediate High

Dextrose equivalent (DE ) 42 52 69

0
Commercial Bauma( B e ) 43O 43O 43O

~ l u c o s e (961 20 28 41
I

Maltose (96) 15 19 22

TABLE, 9.

PROPERTIES OF ACID ENZYME CONVERTED GLUCOSE SYRUP

Dextrose equivalent (DE) 42 52 69

0
Canmercial Baume ( Be' 42 42 42

S o l i d s (%) 80 . 82.2 80

. ~ s ' h(%I 0.65 0.65 0.65

Glucose (%) 23 31 45

Maltose (%) 13 13 20
 CHAPTER 5

SUMMARY AND CONCLUSIONS

5.

ConvenUonally, malt syrups used by food i n d u s t r i e s are produced f r a n

buley. he broad objective of the present investigation was t o produce

rnalt Byrups p r i c i p a l l y fran sorghum and millet grains which would be used

by food industries. T h e Nigerian ecosystan does n o t 8upj$?rt th0 a g r i c u l t w a l

production of barley grains, thus j u s t i f y i n g thL8 objective. The r e s u l t s

prcrduced fran the i n v e s a g a t i o n are summarized as follows:

1. Both the malting and m a l t ' s q u a l i t y c h a r a c t e r i s t i c s of t h e g r a i n s

studied i n d i c a t e t h a t aarghum generates b e t t e r m a l t than millet.

2. 50 hours of steeping and 5 days of gemination a t room temperature gave,

,.,
A*&.- --La --.... d i a s t a t i c power development
YYLY16U111 i n sorghum and m i l l e t grains.
b
3. Feasible ranges of 25-35% s u b s t r a t e and 0.10-0.15% enzyme concentrations

are b e s t s u i t e d f o r syrup production.

4. Sorghum s t a r c h is more adequate f o r glucose syrup prodyction. High

colour value of the syrup obtained could be reduded when i t i s f u r t h e r

refined with carbon and ion-exchange de-ionized processes.

One would therefore suggest f u r t h e r research in comection with,theae

r e s u l t s a s follows:

. More malting 'study is advocated in order t o f i n d b e t t e r favourable

Y

malting conditions f o r m i l l e t ' e s p e c i a l l y i n t h e area of malting l o s s reduction.

b. The c o n t r o l and measurement of colour/flsvol.~,r acquired by sorghum m d l b

during the k i l n i n g process.

c. I s o l a t i o n , p u r i f i c a t i o n ,and characterization of sorghum malts alpha

 . 85,

amylase and i t s tiultability i n starch liquefaction process of glucose syrup

production comparad with that o f a microbial alpha amylase.

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 APPENDIX

Preparation of Analytical reagents, D i n i t r o s a l i c y l i c acid(DNS) reagent .

NaOH, T h i s
-
I g of 3 ' , 58
was made
d i n i t r o s a l i c y l i c a c i d w a s disolved I n 20ml ob 2N
up to 50ml with d i s t i l l e d water. 3 0 9 of potassium
sodiumtartarate (Rochelle S a l t ) was then dissolved i n t h e r e s u l t i n g
s o l u t i o n , and made up t o 10Oml with d i s t i l l e d water and i s c a l l e d DNS
reagent.

Anthrone Reayen t.

7601111 of sulphuric acid (98% W/W H2SC4) Y" &d=d ' ,336n1 of w ~ t e r .
with s t i r r i n g a f t e r cooling, 1g of thiourea and l g of anthroha were added
and s t i r r e d u n t i l dissolved. The r e s u l t i n g s o l u t i o n was s t o r e d i n a
refrigerator.

-
Hydrogen ~ e r o x i d e ( H ~ S O 0.75%
~), Solution:

A f r e s h s o l u t i o n was prepared each time by d i l u t i n g S m l of 30% 4 0 2

(100 v o l ) to 200 m l with d i s t i l l e d water, The r e s u l t i n g s o l u t i o n was-
stored i n a r e f r i g e r a t o r .

Methylene blue, 1%

1g of the pure.dye was dissolved i n l O O m l of d i s t i l l e d water.

Ihver t sugar standard s o l u t i o ~

To 9.50g of pure sucrose was added 5ml of hydrochloric acid and

d i l u t e d with d i s t i l l e d water to 100ml. This w a s s t o r e d a t room temperature
f a r s e v e r a l days, and then made up to 2501111. When required, 25ml of the
stock s o l u t i o n was n e u t r a l i s e d with ocdi-at hydroxide and made up to 200ml
t o give a 0.5% i n v e r t solution.

Fehlinaos Solution A and Br

Commercial Fehling's s o l u t i o n s A and B for laboratory a n a l y s i s were

purchased from BDH company. Before each a n a l y s i s an equal mixture of the
two were made up to 5ml f o r d i a s t a t i c power determination and 25ml f o r
r e d u c i n g / ~ e x t r o s e equivalent determination.

Benedictc s r e a g e n t

1739 of sodium c i t r a t e and lOOg of sodium carbonate were'dissolved i n

8001111 of w m water, f i l t e r e d and made up t o 850ml with water. ~ e a n w h i l e ,
17,3g of Copper Sulphate was dissolved in about l O O m l of water and made up
t o 150ml. The f i r s t s o l u t i o n was poured i n t o a 2 - l i t r e beaker and t h e
Copper Sulphate s o l u t i o n added slowly with stlrrlng.
Acetic acid 1.ON:

28.65m1 g l a c i a l acetic acid a t 2 0 O ~was d i l u t e d t o 5001nl with d i s t i l l e d

water.

Acctate b u f f e r s o l u t i ~ n ,pH 4.6;

689 odium acetat. (CH CalNa.3H20) was dissolved i n 3 0 h l s f L O N

a c e t i c acid and made up to ane l i t r a with d i s t i l l e d water a t 20 C.

Sodium hydroxide, 0. Wt

4-09 of N ~ O Hwas dissolved i n one l i t r e of d i s t i l l e d water.

oy"
2% s t a r c h solution, buffered a t pH 4.6:

A 109 cieam of soluble s t a r c h was made with d i s t i l l e d water.and

poured i n t o about, 400ml of b o i l i n g d i s t i l l e d water s t i r r e d constantly,
he s o l u t i o n was boiled f o r 2 minutes and cooled under cover, t o avoid s k i n
formation t o 20'~. l O m l of a c e t a t e b u f f e r w a s then added and the volume
made up to 500ml. This w a s prepared daily.

0. W ammonia solution:

6.7 m l of the concentrat& ammonia was d i l u t e d t o l l i t r e with d i s t i l l e d

water.

Buffer Preparations:

Buffer were prepared according to t h e method described by Sidney and '

Nathan( 1955 ) as follows:

From the stock s o l u t i o n s of A t 0.l.M a c e t i c acid (5.76m1 i n l Q O O m l water)

and 8: 0.1M sodium a c e t a t e 8.2g In l O O O m l water). Acetate buffer were
prepared thus:
pH 4 t 41.0ml A + 901111B d i l u t e d to l O O m l
pH 5 t 14.8ml A + 35.2m1 B d i l u t e d to l O O m l i

Also f r a n t h e stock s o l u t i o n of A : O.lM monobasic sodium phosphate

(13.99 i n l O O O m l water) and B, 0.lM d i b a s i c s o d i m phosphate 27.83g i n
1000ml water). Phosphate b u f f e r s were prepared thus:

pH 6 s 87.Tml A + 12.3ml B d i l u t e d to l O O m l
pH 7 : 39.01111 A + 61.0ml B d i l u t e d to l O 0 m l
pH 8 t 5.3ml A + 94.7ml B d i l u t e d to 100ml. !,- . .
--+
,*
, ' ,-
"

* \+b
! . I : >