Die Nahrung 32 (1988) 8,783-788 Department of Food Science and Technology, Universify of Nigeria, Nsukka, Nigeria

Estimation of the biological value of food proteins by a modified equation of the essential amino acid index and the chemical score
A. I. IHEKORONYE

A modified essential amino acid index equation incorporating a digestibility factor (MEAAI), and a chemical score index (CSI) were developed from the essential amino acid profile of 18 different food proteins hydrolysed by the combined but sequential action of papain and pronase E enzyme systems. The essential amino acid composition of whole egg was used as a reference standard. The following equations were proposed:

where a-h are the respective values of each of the 8 essential amino acids in the sample; DFp isthe digestibility factor for food protein sample; DF, is the digestibility factor for whole cooked egg; I OOa CSI = ~, % in which a is the value of the limiting amino acid in the sample, and ( is the corresponding value in whole egg. I , Both equations were used to calculate the biological value of different protein foodstuffs. There was a close agreement between values obtained using the equations and reported values for the same foodstuffs published in the literature. There was also a close correlation (r = 0.665) between data obtained using MEAAI and those obtained using CSI in estimating biological values, suggesting that either equation could be used reliably and solely to estimate protein biological value of foods.

Introduction

It has been recognized since the investigations of OSBORNE al. [I] that the nutritional et value of a protein is primarily dependent upon its constituent amino acids. MENDEL [2] proposed that the efficiency of the individual protein in this respect must depend on the minimum quantity of any indispensable amino acid that it will yield. On the basis of this concept, MITCHELL al. [3] evolved a workable system for the quantitative evaluation of a et protein from its amino acid composition. Using whole egg protein as the standard, the nutritive value of a protein was expressed as a chemical score equal to the greatest percentage deficit in an essential amino acid in the test protein. OSER devised a similar method [4] in which all the essential amino acid egg ratios were integrated by calculation of their geometric mean. With reference to man and the rat both methods yielded values which correlated well with the biological values of a variety of proteins. Thus defined, it is possible to calculate the protein value of foods. What is logically re-

784

Die Nahrung 32 (1988) 8

quired are figures portraying the amino acid content and availability. In theory a combination of these two factors, availability and digestibility, will overcome the disadvantages of chemical and in vivo biological methods. Yet, in so doing, the problem lies in the most convenient and accurate method which should be used for protein hydrolysis, quantitation and digestibility determination Earlier experiments had reported on methods for predicting the in vivo rat-based protein digestibilities from in vitro enzymic and chromatographic procedures [5, 61. Techniques for complete hydrolysis of proteins using enzymes and subsequent accelerated quantitation of released amino acids were parts of such reports. A multiple regression equation for predicting the in vivo rat-based protein efficiency ratio had been developed from amino acid profiles provided by enzymically released amino acids [7]. Hence the present study was undertaken to evaluate whether an incorporation of a digestibility factor into OSER'S essential amino acid index [4] would predict the in vivo based biological value of food proteins. It should also answersthequestion whether chemical score and essential amino acid indices derived from enzymically released amino acid data could be used to evaluate protein nutritional quality of foods and foodstuffs.

Material and methods

Commercial and laboratory prepared samples consisting of plant, animal and mixed protein diets were evaluated. These had been described in earlier reports 15-71 and consist essentially of the samples shown in Table 1.

Table I Percent protein and KJELDAHL factors for food samples analyzed
Food or food ingredient Protein [ %I 45.8 83.8 7.40 48.4 9.69 90.6 48.2 19.8 14.3 21.6 16.8 19.7 18.7 56.5 20.9 46.4 61.5 64.6 60.0

Whole cooked egg Cooked chicken muscle Corn meal Soy flour 20 PDI Shredded wheat ANRC casein Textured vegetable protein Life cereal Cheerios Fish fillets Macaroni and cheese dinner Chicken and dumpling Breakfast bars Cottonseed meal Beef noodle dinher Lean beef Peanut flour SR-57 Peanut flour pearl Peanut flour C-32W

KJELDAHL factor ________ 6.25 6.25 5.70 5.71 5.70 6.38 5.71 6.25 5.83 6.25 6.25 6.25 6.25 6.25 6.25 6.25 6.25 6.25 6.25

IHEKORONYE:Biological value of proteins

785

Essential atnino acid index

OSER developed an integrated index where he used the geometric mean of the egg ratios to estimate the [4] biological value of a protein. An essential amino acid index (EAAI) was then defined as the ratios of the essential amino acids in a protein relative to their respective amounts in whole egg protein. Precisely,
n-x

lOOa
~

lOOb be

ae

-x
ce

lOOc

1OOh

he

in which a, b, c ... h are percent of each of then essential amino acids in the food protein, and up. b,, ce, ... he are percent of the respective amino acids in the whole egg protein. The computation can be conducted logarithmically thus:

It is recognized that the essential amino acid index evaluates the prowin when fully digested and therefore tends to over-estimate the true biological value of the protein. Bui i111\ condition can be ameliorated by including a digestibility value derived under the same experimental conditions as the amino acids used in the equation. The suggested equation is a modification of the essential amino acid index and results in a modified essential amino acid index equation:

where a-h = concentration (mg/16 mg N) of each of the essential amino acids (leucine, isoleucine, lysine, threonine, tryptophan, valine, phenylalanine + tyrosine, cystine + methionine) in protein sample, DFp = digestibility factor for protein sample, DF, = digestibility factor for whole cooked egg. MEAAI was compared to available biological value data reported in the literature for samples studied.

Chemical score indelc ( C S I )

The levels of the essential amino acids and determined by enzyme hydrolysis and GLC in the food proteins were compared with those in whole cooked egg protein. The essential amino acid of the sample which had the greatest percentage deficit compared with the amino acid in the whole cooked egg protein was used to calculate the chemical score index thus: 1OOa CSI =
~

where a = mg/16 mg N of essential amino acid in food protein, ae = mg/16 mgN of essential amino acid in whole cooked egg protein.

a,

Results and discussion

Modified essential amino acid index (MEAAI) OSERS essential amino acid index [4] was modified to include the digestibility factor obtained for each protein foodstuff by the papain-pronase E colourimetric procedure. The calculated essential amino acid indexes for various food proteins are presented in Table 2.
SO

Die Nahrung 32 (1988) 8

786
Table 2 Modified essential amino acid indexes of various food proteins using eight essential amino acid
Food proteins
Whole cooked egg Cooked chicken muscle Soy flour 20 PDI ANRC casein Textured vegetable protein Life cereal Cheerios Fish fillets Macaroni and cheese Chicken dumpling Breakfast bars Cottonseed meal Beef noodle Lean beef
a

Die Nahrung 32 (1988) 8

Modified essential amino acid index 100.0 73.5 69.1 61.9 66.9 67.8 70.3 56.9 69.2 6@3 61.7 59.7 72.2 62.3

Biological value 100.0 74.3b 72.8b 79.7b 67.6b 71.8b

66.0b 74.3b

The eight essential amino acids included in all computations are: lysine, tryptophan, isoleucine, leucine, valine, methionine plus cysteine. threonine and phenylalanine plus tyrosine Food and Agricultural Organization [8]

These values were compared to available biological value data for the respective proteins reported in the literature. The MEAAI values correspond very well to biological values reported by F A 0 [8] for cooked chicken muscle, soy flour, textured vegetable protein, fish fillets and cottonseed meal. Differences in values between MEAAI and reported biological values may be due to dissimilarities in composition and protein content of materials compared. For instance, the biological value of lean beef was reported to be 74.3 whereas the MEAII calculated it to be 62.2, a discrepancy which may have arisen from differences in the amount of lean and digestibility of the samples evaluated.
Clienzicnl score index

Table 3 presents the chemical score of the food proteins calculated with the equation together with the limiting amino acids as quantitated from enzymatic hydrolysis of the proteins. The most common limiting amino acids in all protein foods evaluated were either the sulphur-containing amino acids, lysine and threonine. The chemical scores computed were based on the second limiting amino acid since the values for threonine were consistently lower in each sample. The sulphur containing amino acids and lysine were found to be the second limiting amino acids for several samples while lysine, tyrosine and phenylalanine were used in others. Calculation of chemical scores with the enzymically released essential amino acids using amino acid composition of whole egg as a reference standard produced results in agreement with values reported [S] for casein, soy flour and durum wheat flour. The two indices were compared as tools for evaluating protein quality. There was a significant relationship (r = 0.665) between MEAAI and CSI, suggesting that either index could be used to calculate the expected biological value of food proteins.

IHEKORONYE: Biological value of proteins

787

Table 3 Calculated chemical score of food proteins using whole cooked egg as the reference pattern
Food products Chemical score
1O(r.0

Limiting amino acid

Whole cooked egg Cooked chicken muscle Corn meal Soy flour 20 PDI Shredded wheat Textured soy protein ANRC casein Textured vegetable protein Life cereal Cheerios Durum wheat flour Fish fillets Macaroni and cheese Chicken dumpling Breakfast bars Cottonseed meal Beef noodle Leaf beef

58.2 19.0 55.8 52.8 61.0 80.6 61.0 56.8 48.3 39.0
50.6

50.1 53.2 21.7 58.2 89.0 61.4

cystine and methionine lysine cystine and methionine lysine, threonine methionine and cystine leucine methionine and cystine cystine and methionine lysine, threonine lysine, threonine tyrosine, phenylalanine lysine methionine and cystine phenylalanine, tyrosine threonine lysine methionine and cystine

Protein nutritional value is a combination of two factors; the total essential amino acid content of the protein, and protein digestibility which is an indicator of the availability of the essential and non essential amino acids. Thus, the use of adjustments in a predictive equation for protein nutritional value which incorporate these factors will lead substantially to increased estimates of protein nutritional quality. The protein nutritional indices proposed in the present study produced results in agreement with in vivo derived values reported for the foodstuffs, hence would be good estimates of the expected in vivo biological value of food proteins. The modified equations have been tested against independent literature data and found to agree closely with measured values.
Zusammenfassung
A . I . IHEKOKONYE:

Bestimmung der biologischen Wertigkeit von Nahrungsproteinen durch eine modifizierte Gleichung des ,,essential amino acid index" und des ,,chemical score''

Aus dem Profil der essentiellen Aminosauren von 18 verschiedenen Nahrungsproteinen, die nacheinander mit Papain und Pronase E hydrolysiert wurden, wird eine modifizierte Gleichung des .,essential amino acid index" einschliel3lich eines Verdauungsfaktors (MEAA) abgeleitet. Die Zusammensetzung der essentiellen Aminosauren von Vollei wird als Bezugswert verwendet. Folgende Gleichungen werden vorgeschlagen :
1 lOOa DF lOOb DF lOOh DF log MEAAl = -log x 2 + log x 2 + . . . + log - x ' 8 a, DF, be D.Fe he DF, a-e sind die jeweiligen Werte der 8 essentiellen Aminosauren in der Probe; DFp ist der Verdaulichkeitsfaktor fur Nahrungsprotein-Probe; D F . ist der Verdaulichkeitsfaktor fur Vollei; 1OOa CSI = __
~ ~

worm a der Wert der limitierenden Aminosaure der Probe und a, der entsprechende Wert von Vollei ist.

a,

788

Die Nahrung 32 (1988) 8

Beide Gleichungen dienen zur Berechnung der biologischen Wertigkeit verschiedener Proteinnahrungsmittel. Es besteht eine enge ubereinstimmung zwischen den nach diesen Gleichungen berechneten und Literatur-Werten der gleichen Nahrungsmittel. Die zur Bestimmung der biologischen Wertigkeit erhaltenen MEAAI-Werte korrelieren gut mit den berechneten CSI-Werten ( r = 0,665). Daraus geht hervor, daB jede Gleichung zuverlassig und a k i n zur Bestimmung der biologischen Wertigkeit von Nahrungsproteinen herangezogen werden kann.

A. M.MXEKOPOHbE OllpeneJIeHHe 6 e o n o r ~ r e c ~ oUeHHOCTH IlAUleBOTO 6 e n ~ a ii C BaHHoro ypaaHeHaa essential amino acid index H chemical score.

IIOMOUlbK)

MOnH@HUApO-

M3 npOl$Hnfl He3aMeHHMbIX aMHHOKHCfiOT 18 pa3JlHYHbIX nlllUeBbIX 6en~o6, KOTOPbIe OnHH 3a nPYrHM r ~ n p o n ~ s a p o ~ a nnanamoM H npoaasoii E, BbiBonHTcr Mo&xiqtiposaHHoe ypaBHeHwe essential ~cb amino acid index BKJItOYHTenbHO nHueBapHTenbHor0 @aKTOpa(MEAA). COCTaB He3aMeHHMbIX BMHHOB KMcnoT rii4a ~cnonb3ye~cnKaYecTse cTaHnaprHol senwimu. IIpennaramTcr cnenytouae ypaeHeHm : 1 lOOa DF 1006 DF lOOh log MEAAI = - log __ x 2 + log __ x 2+ log __ x 8 a, DF, 6 , DF, he DF,

References

[I] OSBORNE, B., L. B. MENDEL E. L. FERRY. biol. Chem. 46, 1645 (1919). J. and J. [2] MENDEL, B., Nutrition: The Chemistry of Life, Yale University Press, New Haven 1923. L. [3] MITCHELL, H., and R. J. BLOCK, biol. Chem. 163, 599 (1946). H. J. [4] OSER,B. L., J. am. dietet. Assoc. 27, 396 (1951). (51 IHEKORONYE, A. I., J. sci. Food Agric. 36, 1004-1012 (1985). [6] IHEKORONYE,I., J. Food Technol. 21, 81-87 (1986). A. [7] IHEKORONYE,I., Ph. D. Thesis, University of Missouri, Columbia 1981. A. [ ] FOOD AND AGRICULTURAL ORGANIZATION, Protein Requirements, F A 0 Nutr. Meet. Rep. 37. 8 FAO, Rome 1983.
Department of Food Science and Technology, University of Nigeria, Nsukka, Nigeria Dr. A. I. IHEKORONYE, Received March 17, 1987