Plantas aromticas

Reconhecer os aspectos biolgicos diferenciadores das plantas aromticas

Reconhecer as caractersticas fsico-qumicas genricas dos metabolitos secundrios
caractersticos
Conhecer os principais mtodos de anlise e os seus fundamentos

Reconhecer a importncia das plantas aromticas enquanto matria-prima Farmacutica

Reconhecer plantas aromticas e leos essenciais como ingredientes activos de medicamentos
Conhecer efeitos adversos associados administrao de extractos aromticos

Farmacopeia Portuguesa VIII (2006)

Frmaco
inscrito

Descrio

Taxon de origem

Serpo

Partes areas floridas, secas, inteiras ou

fragmentadas.

Thymus serpyllum L

Tomilho

Folhas e flores inteiras, destacadas dos ramos.

Absinto

Folhas basilares ou inflorescncias ligeiramente

folhadas, ou mistura destes rgos, inteiros ou
fragmentados.

Artemisia absinthium L.

Alecrim

Folhas inteiras ou fragmentadas, secas.

Rosmarinus officinalis L.

Eucalipto

Folhas secas, inteiras ou cortadas, dos ramos

mais idosos.

Eucalyptus globulus Labill.

Hortel-pimenta

Folhas secas, inteiras ou cortadas.

Mentha x piperita L.

Melissa

Folhas ou ramos floridos, secos.

Melissa officinalis

Salva

Folhas secas, inteiras ou fragmentadas.

Salvia officinalis L.

Salva-trilobada

Folhas secas, inteiras ou fragmentadas.

Salvia fructicosa Miil.

Thymus vulgariis ou T. zygis

Farmacopeia Portuguesa VIII (2006)

Frmaco
inscrito

Descrio

Taxon de origem

Alfazema

Flores, secas.

Lavandula angustifolia P. Miller

Arnica

Captulos, inteiros ou fragmentados, secos.

Arnica montana L.

Camomila

Captulos secos.

Matricaria recutita L.

Camomila-romana

Captulos secos.

Chammamaelum nobile (L.) All.

Cravinho

Boto floral inteiro.

Syzygium aromaticum (L.)

Flor de laranjeira
amarga

Botes florferos, secos.

Citrus aurantium L. ssp.

aurantium

Serpo

Partes areas floridas, secas, inteiras ou

fragmentadas.

Thymus serpyllum L

Tomilho

Folhas e flores inteiras, destacadas dos ramos,

secos.

Thymus vulgaris ou T. zygis.

Farmacopeia Portuguesa VIII (2006)

Frmaco inscrito

Descrio

Taxon de origem

Alcarvia

Aqunios.

Carum carvi L.

Anis

Diaqunios maduros

Pimpinella anisum L.

Anis estrelado

Frutos mltiplos de folculos, secos.

Illicium verum Hooker fil

Coentro

Diaqunios.

Coriandrum sativum L.

Funcho amargo

Aqunios.

Foeniculum vulgare ssp. vulgare var.

vulgare

Funcho-doce

Aqunios.

Foeniculum vulgare ssp. vulgare var. dulce

Ns moscada

Amndoas do fruto.

Myristica fragans Houtt

Zimbro

Glbulas maduras.

Juniperus communis L.

Frmaco inscrito

Descrio

Taxon de origem

Canela da china

Casca seca.

Cinnamomum cassia Blume

Canela de Ceilo

Casca seca, privada do sber e do

parnquima.

Cinnamomum zeylanicum Nees

Casca de laranja
amarga

Parte mais externa do epicarpo,

seco.

Citrus aurantium L. ssp. aurantium

Curcuma-de-Java

Rizoma cortado em fatias e seco.

Curcuma xanthorrhiza Roxb.

Farmacopeia Portuguesa VIII (2006)

Frmaco inscrito

Taxon de origem

leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo
leo

Rosmarinus officinalis L.
Lavandula angustifolia P. Miller
Pimpinella anisum L.
Illicium verum Hooker fil
Matricaria recutita L.
Cinnamomum cassia Blume
Cinnamomum zeylanicum Nees
Cinnamomum zeylanicum Nees
Syzygium aromaticum (L.) Merril et L. M. Perry
Eucalyptus globulus Labillardire
Citrus aurantium L. ssp. aurantium
Foeniculum vulgare ssp. vulgare var. vulgare Miller
Foeniculum vulgare ssp. vulgare var. dulce Miller
Mentha arvensis L., var. glabrata (Benth.)
Mentha x piperita L.
Citrus sinensis L.
Citrus limon (L.) Burman fil
Melaleuca alternifolia (Maiden et Betch) Cheel
Myristica fragans Houtt
Salva sclarea L
P. pinea L e Pinus pinaster Ait
Thymus vulgaris L e T. zygis Loefl. ex L.
Juniperus communis L

essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial
essencial

de
de
de
de
de
de
de
de
de
de
de
de
de
de
de
de
de
de
de
de
de
de
de

alecrim
alfazema
anis
anis estrelado
camomila
canela da china
canela de Ceilo
Caneleira
cravinho
eucalipto
flor de laranjeira amarga
funcho amargo
funcho-doce
hortel-japonesa
hortel-pimenta,
laranja doce
limo
melaleuca
noz-moscada
salva-esclareia
terebintina
tomilho
zimbro

Massa at 300 Da
Reduzida superfcie polar
Hidrofbicos

Metabolitos volteis

Marcadores distintivos de um cluster bem definido de plantas medicinais

O CH3

a-Pineno

Mirceno

b-Felandreno

Limoneno
Anetol

a-Felandreno

metil-etilsulfito

a-Fencheno

a-Tujeno
para-Cymeno
D-2-Careno

Mentol
OH

OH

OH
OMe

Verbeneno
Canfeno

Sabineno

b-Pineno

1,8-Cineol

Germacreno D

D-3-Careno

Eugenol

Carvacrol

E-Cariofileno

metil-3-metilbutanoato

1-Octeno-3-ol

Aldedo cinmico
OH

CHO

Aristolona

Metabolitos volteis
Marcadores distintivos de um cluster bem definido de plantas medicinais

Rosmarinus officinalis: (SEM)

leos essenciais, leos volteis, Essncias

Misturas complexas de compostos orgnicos volteis extrados de plantas
(aromticas) por destilao ou expresso.

So insolveis na gua apesar de

poderem conter constituintes
parcialmente solveis

ISO 9235 (1997), ISO/TC 54; NP 90 (1987) do IPQ-CT 5

Metodologias e estratgias de anlise de

misturas complexas de compostos volteis

60 min

Analytical strategies

Collections of compounds available for screening, complex mixtures for analysis

LL : P-LC : P-CG : TLC : HPLC-P

CLASSIC APPROACH

Fractioning
Fractioning
Fractioning

MS : IR : NMR

Purification

Spectroscopy

Purification

Spectroscopy

Purification

Spectroscopy

Purification

Spectroscopy

Purification

Spectroscopy

Purification

Spectroscopy

Purification

Spectroscopy

Purification

Spectroscopy

Identity

xN

APPROACH pro-EFFICIENCY
HR Separation / Spectroscopy

Towards efficiency in complex mixtures analysis

Identity

Theory of chromatography (in brief)

Cavaleiro, C.
Group Drug Discovery, CEF
Univ. Coimbra PT

Theory of chromatography (in brief)

Partition (mass transfer)

Mechanisms and partition coefficient
Retention
Retention volume and retention time
Dead volume e dead time
Adjusted retention time
Capacity factor
Selectivity

Efficiency
Van Deemter equation
Plate count
Plate height

Resolution

Theory of chromatography (in brief)

Retention induced by the partition of solutes between stationary and

mobile phases (SP and MP)

Partition coefficient (K)

[ solute]in SP
[ solute]in MP
Qs

Vs Vm .Qs
[ solute]in SP

[ solute]in MP Qm
Vs .Qm
Vm

Vm Qs
.
Vs Qm

Capacity factor (K)

or

K b .k

K is specific of each solute for a given column and stationary phase and
directly related with the retention

Theory of chromatography (in brief)

Capacity factor K correlation with retention time Tr

Qs
k
Qm

Qs t r t d

k
Qm
td

tr td
k
td

td

Time

Selectividade
Theory of chromatography (in brief)

t r1 t d
k1
td

tr 2 td
k2
td

tr 1
td
tr 2
Tempo

k2 tr 2

a
k1 tr1

Selectivity

Theory of chromatography (in brief)

Towards to efficiency

Theory of chromatography (in brief)

Plate Height (H) and Plate count (n)

Theory of chromatography (in brief)

Van Deemter equation

B
H A Cu x
ux
A Multiple path effect (Eddy diffusion)
B Longitudinal diffusion
C Mass transfer kinetics
ux Linear speed of the mobile phase

Theory of chromatography (in brief)

L/H=n
H=L/n

Time

Time

Towards to efficiency

Theory of chromatography (in brief)

Selectivity

a = k2 / k1

Time

Resolution
R

Time

N a 1
k2
.
.
4
a
(1 k2 )

Towards to efficiency

Theory of chromatography (in brief)

Summary

Towards efficiency:
- the narrowest column
- the smallest phase ratio (b

- the slimmest bed phase and regular surface

- the longest column
- suitable phase composition

- the suitable mobile phase and flow

Theory of chromatography (in brief)

Summary

Towards efficiency:
- the narrowest column
- the smallest phase ratio (b

- the slimmest bed phase and regular surface

- the longest column
- suitable phase composition

- the suitable mobile phase and flow

Complex volatile mixtures analysis and molecular characterization

Gas chromatography (GC)

and

derived techniques

Applied to essential oil analysis

Gas chromatography: hardware

Mobile phase: Gas (Inert, low viscosity and low compressibility)

Stationary phase:
Adsorptive solid Gas-solid chromatography (adsorption)
Thermo stable liquid - Gas-liquid chromatography (partition)
Compatible samples: Gases or volatile samples

Gas chromatography: hardware

Packed columns
- Stainless still or glass, 1 - 4 mm i.d., 3 - 4 m long.
- Pack: frit of inert particles (coated)

Capillary columns (high resolution columns)

- Glass or fused silica, 0,1 0,4 mm i.d., 10 a 200 m long.
- Wall coated (WCOT)

100% dimetil polisiloxano

Estvel at 360C
Polaridade: no polar
Uses: solventes, deriv. petroleo,
medicamentosas, ceras
6% cianopropilfenil - 94% dimetil polisiloxano
Estvel at 280C
Polaridade: ligeiramente polar
Usos: Terpenoides volteis, insecticidas,
residuos de solventes em produtos
farmaceuticos
5% difenil - 95% dimetil polisiloxano
Estvel at 360C
Polaridade: no-polar
Usos: aromas, anlises ambientais,
hidrocarbonetos aromaticos.
35% difenil - 65% dimetil
polisiloxano
Estvel at 300C
Polaridade: polaridade intermdia
Usos: pesticidas, Aroclors, aminas
herbicidas azotados
20% difenil - 80% dimetil
polisiloxano
Estvel at 310C
Polaridade: ligeiramente polar
Usos: terpenoides volteis, alcoois
50% fenil - 50% metil polisiloxano
Estvel at 340C
Polaridade: polarid. intermdia
Usos: trigliceridos, ftalatos, esteroides,
fenois

14% cianopropilfenil - 86% dimetil

polisiloxano
Estvel at 280C
Polaridade: polaridade intermdia
Usos: pesticidas, alcoois, oxigenados
trifluoropropilmetil polisiloxano
Estvel at 360C
Polaridade: selectiva para para comp com
pares de electres desemp.
Usos: anlises ambientais, solventes, Freons,
medicamentos, cetonas, alcoois
65% difenil - 35% dimetil polisiloxano
Estvel to 370C
Polaridade: polaridade intermdia
Usos: trigliceridos, acidos, acidos gordos
livres
90% biscyanopropil - 10%
cianopropilfenil polisiloxano
Estvel at 275C
Polaridade: muito polar
Usos: FAMEs, cis/trans e smeros de
dioxinas
Carbowax PEG
Estvel at 250C
Polaridade: polar
Usos: FAMEs, aromas, acidos, aminas,
solventes, xileno isomeros
50% cianopropilfenil - 50% fenilmetil
polisiloxano
Estvel at 260C
Polaridade: polar
Usos: FAMEs, carbohidratos

Gas chromatography: hardware

Running operation
Mobile phase
Flow and pressure control
Sampling
Injection type (split, splitless, o.c.)
Vaporization chamber
Oven temperature
Detection
Universal detectors / Selective detectors / Spectroscopic detectors

Gas chromatography: hardware

He
Ar K H2

N2

Gas chromatography: hardware

Gas chromatography: hardware

Detectors
Detector

Type

Support

Selectivity

Sensitivity

Linear.
Range

Flame ionization
(FID)

Mass flow

Hydrogen and
air

Most of organic compounds

100 pg

107

Thermal conductivity
(TCD)

Concentration

Reference

Universal

1 ng

107

Electron capture
(ECD)

Concentration

Make-up

Halids, nitrates, nitrides, peroxide,

anhydrides, organmethalics

50 fg

105

Nitrogen-Phosphorous
(NPD)

Mass flow

Hydrogen and
air

Nitro and phosphorous organic

compounds

10 pg

106

Flame photometry
(FPD)

Mass flow

Hydrogen and
air
(Oxygen)

S, B, As, Ge, Se, Cr organic

compounds

100 pg

103

Make-up

Aliphatic ,aromatic, ketones,

esteres, aldehyds, amines,
heterocycle, tioorganic

2 pg

107

Photo-ionization
(PID)

Concentration

Gas chromatography: hardware

Detectors

Gas chromatography: hardware

Detection
(FID)

eV

Time

eV

FID

Gas chromatography: improving separation

Oven temperature: Isothermal operation

tr td
Q
k s
td
Qm
Isothermal 80 C

Isothermal 150 C

Qe and Qm depend of temperature

- solubility in the phase
- vapour pression

Gas chromatography: improving separation

Oven temperature: Thermal gradient

tr td
Q
k s
td
Qm

Qe and Qm depend of temperature

- solubility in the phase
- vapour pression

150

80

80 C (10 min), 5 C / min (150 C), 10 min

80 C, 3 C / min (150 C)

Gas chromatography: the chromatogram

A
b
u
n
d
a
n
c
e
T
I
C
:J
A
L
P
I
N
A
B
.
D

4
0
0
0
0
0

3
5
0
0
0
0

3
0
0
0
0
0

2
5
0
0
0
0

2
0
0
0
0
0

1
5
0
0
0
0

1
0
0
0
0
0

5
0
0
0
0

0
5
.
0
0
1
0
.
0
0
1
5
.
0
0
2
0
.
0
0
2
5
.
0
0
3
0
.
0
0
3
5
.
0
0
4
0
.
0
0
4
5
.
0
0
5
0
.
0
0
5
5
.
0
0
6
0
.
0
0
T
i
m
e
>

The chromatogram

Data File : Z:\HPCHEM\1\DATA\ROSOF837.D

Vial: 1
Acq On : 23 Apr 03 10:54
Operator: Cavaleiro
Sample : Rosmarinus off 837
Inst : GC/MS Ins
Misc
: CASAAN
Multiplr: 1.00
Sample Amount: 0.00
Method : G:\HPCHEM\3\METHODS\ENVDEF.M
Title :
Signal : FID
peak
R.T.
PK
peak
raw..
#
min
TY
height
area.
--- ----- ----- ---- ---- --- ------- ------- ------- ------ ------1
2.648
BB 3
151546
2894980
2
3.123
BV
252982
5086949
3
3.235
VV
396428
7594596
4
3.392
VB 3
88935
2442273
5
4.440
BV 2
241923
5259853
6
4.534
VV
321728
7083312
7
4.575
VV
279492
7072723
8
4.762
PB
1546004
486326208
9
5.001
BB
6329338
150354755
10
5.338
BV
808109
20554003
11
5.406
VV 3
518081
15033680
12
5.570
VB
4411881
111773185
13
5.870
BV
13236279
579112549
14
6.169
BB
433333
10763430
15
6.480
VV 2
829160
22600669
16
6.558
VB
2622491
74594717
17
6.809
BV 4
10332165
470719718
18
6.940
VB 3
76024
2175548
19
7.591
BV
2243092
59312870
20
7.692
VB 3
115337
3271280
21
8.287
BV 5
65276
2350983
22
8.489
VV
611445
17781904
23
8.695
VB 2
3462224
127629629
24
9.129
BV 2
363352
1192061
25
9.290
PV 4
74423
2990871

% of
total
------0.087%
0.156%
0.239%
0.070%
0.165%
0.226%
0.206%
1.577%
4.851%
0.659%
0.461%
3.607%
18.716%
0.336%
0.725%
2.345%
15.214%
0.062%
1.909%
0.099%
0.071%
0.569%
4.091%
0.369%
0.090%

peak
R.T.
PK
peak
raw..
#
min
TY
height
area.
--- ----- ----- ---- ---- --- ------- ------- ------- ------ ------26
9.648
BB 3
151546
2894980
27
10.123 BV
252982
5086949
28
10.235 VV
396428
7594596
29
10.392 VB 3
88935
2442273
30
11.440 BV 2
241923
5259853
31
11.534 VV
321728
7083312
32
11.575 VV
279492
7072723
33
11.762 PB
15460041
486326208
34
12.001 BB
6329338
150354755
35
12.338 BV
808109
20554003
36
12.406 VV 3
518081
15033680
37
12.570 VB
4411881
111773185
38
12.870 BV
13236279
579112549
39
13.169 BB
433333
10763430
40
13.480 VV 2
829160
22600669
41
14.558 VB
2622491
74594717
42
14.809 BV 4
10332165
470719718
43
14.940 VB 3
76024
2175548
44
15.591 BV
2243092
59312870
45
16.692 VB 3
115337
3271280
46
17.287 BV 5
65276
2350983
47
17.489 VV
611445
17781904
48
18.695 VB 2
3462224
127629629
49
19.129 BV 2
363352
11692061
50
19.290 PV 4
74423
2990871
51
14.558 VB
2622491
74594717
52
14.809 BV 4
10332165
4707197
53
14.940 VB 3
76024
2175548
54
15.591 BV
2243092
59312870
55
16.692 VB 3
115337
3271280
56
17.287 BV 5
65276
2350983
57
17.489 VV
611445
17781904
58
18.695 VB 2
3462224
127629629
Sum of corrected areas: 7091981135

% of
total
------0.087%
0.156%
0.239%
0.070%
0.165%
0.226%
0.206%
15.717%
4.851%
0.659%
0.461%
3.607%
18.716%
0.336%
0.725%
2.345%
15.214%
0.062%
1.909%
0.099%
0.071%
0.569%
4.091%
0.369%
0.090%
2.345%
1.214%
0.062%
1.909%
0.099%
0.071%
0.569%
4.091%

Fictitious chromatogram

Area Percent Report

The chromatogram

Abundance

4.7
5.0
60

6000000

5.87

TIC:
6.81

ROSOF837.D
9.86

5500000
5000000
5.57

4500000
4000000

8.70

3500000

11.97

3000000
6.56
2500000
7.59
2000000
1500000
1000000
500000

5.34
3.24
3.12
2.65 3.39

5.41
4
.5
3
4
.5
7
4
.4
4

11.68
10.80
11.26
10.15

6.48
8.49
9.13

6.17

6.94 7.69
8.29

9
.2
9
9
.3
8

1
0
.5
5
1
0
.4
1

0
3.00

4.00

5.00

6.00

7.00

8.00

9.00

13.18
12
.59
12.1
5
14
13.6
3.11

10.00 11.00 12.00 13.00 14.00

Time-->

Sample complexity
Components quantification
Components identification

Improving methodologies towards components identification in

complex volatile mixtures: GC
Phase a
10

25

15

10

15

20

25

30

RT

Area

10,0

65.000

15,0

60.000

25,0

75.000

total

200.000

35

RT (min)

Sample complexity
Components quantification
Components identification

Reference compounds

?
Retention databases

Improving methodologies towards components identification in

complex volatile mixtures: GC
Is retention time (or ret. volume) an appropriate feature to bank ?
Phase a: Experimental w

TX
Phase a : Experimental r

X
TX
0

10

15

20

25

30

35

Improving methodologies towards components identification in

complex volatile mixtures: GC
Looking for appropriate features to bank Relative retention indices.
Phase a: Experimental w
Z

Z+1

TZ w TX w
0

10

TZ+1 w
15

20

Phase a : Experimental r
Z

25

30

35

Z+1

X
TX r

TZ r
0

10

15

TZ+1 r
20

25

30

35

Improving methodologies towards components identification in

complex volatile mixtures: GC
Looking for appropriate features to bank
Phase a: Experimental Conditions w
Z+1
Z

Retention indices by
linear interpolation

TZ w TX w
0

10

TZ+1 w
15

20

Phase a : Experimental Conditions r

Z

n-alkanes

25

30

35

10

T x T z
I 100 z 100

T
( z 1)
(z)

TX r
15

Fat acids methyl esters

Z+1

X
TZ r

and

TZ+1 r
20

25

30

35

Example: calculation of retention indices for peaks A - D

Consider retention times of the linear alkanes

n-nonane
4.11

n-decane
6.21

n-undecane
9.08

n-dodecane
12.82

Abundance
TIC: BASI.D
9.78
1200000
1100000
1000000

900000

11.49

4.72

800000
700000
600000
500000
3.59
400000

11.09

A
C

300000

D
Polydimethylsiloxane
DB-1 column

200000
100000
0
3.00

4.00

5.00

6.00

7.00

8.00

9.00

10.00 11.00 12.00 13.00 14.00 15.00

Example: calculation of retention indices for peaks A - D

Computing retention index for peak A:

T x T z
I 100 z 100

T
( z 1) ( z )

4,72 4,11
I 900 100
929

6,21 4.11

Abundance

1000000

929

900000
4.72

800000

1164

n-dodecane

1100000

TIC: BASI.D
9.78

n-undecane

1200000

n-decane

n-nonane

1119

11.49

700000

1154

600000
500000
3.59
400000

11.09

300000

200000

Polydimethylsiloxane
DB-1 column

100000
0
3.00
Time-->

4.00

5.00

6.00

7.00

8.00

9.00

10.00 11.00 12.00 13.00 14.00 15.00

Example: calculation of retention indices for peaks A - D

Quest RI database for identities:

D
COOCH3

OH

OH

n-undecane

1100000

n-decane

n-nonane

1200000

TIC: BASI.D
9.78

929

1000000
900000

4.72

800000

1164

n-dodecane

1119

Abundance

11.49

700000
600000

1154

500000
3.59
400000

11.09

300000

200000

Polydimethylsiloxane
DB-1 column

100000
0
3.00
Time-->

4.00

5.00

6.00

7.00

8.00

9.00

10.00 11.00 12.00 13.00 14.00 15.00

Improving methodologies towards components identification in

complex volatile mixtures: GC
Phase a
10

25

15

10

15

20

25

30

RT

Area

10,0

65.000

15,0

60.000

25,0

75.000

total

200.000

35

RT (min)

Sample complexity
Components quantification
Components identification

Databases of retention data

Improving methodologies towards components identification in

complex volatile mixtures: GC
10

Phase a

25

15

10

15

20

25

30

22

7,5
17

10

15

20

25

30

RT

Area

10,0

65.000

15,0

60.000

25,0

75.000

total

200.000

RT

Area

7,5

45.000

12,5

60.000

17,0

20.000

22,0

75.000

total

200.000

35

Phase b
12,5

35

Improving methodologies towards components identification in

complex volatile mixtures: GC

Summary
Chromatographic running on two phases:
- consolidates information about sample complexity
- improve quantification process
- duplicates qualitative information (retention data)

Components identification

Databases of retention data for

multiple phases

Improving methodologies towards components identification in

complex volatile mixtures: GC
10

Phase a

25

15

10

15

20

25

30

22

7,5
17

10

15

20

25

30

RT

Area

10,0

65.000

15,0

60.000

25,0

75.000

total

200.000

RT

Area

7,5

45.000

12,5

60.000

17,0

20.000

22,0

75.000

total

200.000

35

Phase b
12,5

35

Improving methodologies towards components identification in

complex volatile mixtures: GC
10

Phase a

Phase a - database response

25

15

IRa=1020

10

15

20

25

30

35

Phase b

Phase b - database response

IRb=1205

IRb=1215

22

12,5
7,5
17

IRa=1020 IRb=1205
0

10

15

20

25

30

35

IRa=1020 IRb=1215

Improving methodologies towards components identification in

complex volatile mixtures: GC
10

Phase a

Phase a - database response

25

15

IRa=1020

10

15

20

25

30

35

Phase b

Phase b - database response

IRb=1205

IRb=1215

22

12,5
7,5
17

IRa=1020 IRb=1205
0

10

15

20

25

30

35

IRa=1020 IRb=1215

Improving methodologies towards components identification in

complex volatile mixtures: GC
Re
sp
o
n
se
_

2x GC (RI)

Quantification

T
HYM
AST
B.D\F
ID2
B

1
7
0
0
0
0
0
1
6
0
0
0
0
0
1
5
0
0
0
0
0
1
4
0
0
0
0
0
1
3
0
0
0
0
0
1
2
0
0
0
0
0
1
1
0
0
0
0
0
1
0
0
0
0
0
0
9
0
0
0
0
0
8
0
0
0
0
0
7
0
0
0
0
0
6
0
0
0
0
0
5
0
0
0
0
0
4
0
0
0
0
0
3
0
0
0
0
0
2
0
0
0
0
0

R
e
s
p
o
n
s
e
_

1
0
0
0
0
0
5
.0
0

1
0
.0
0

1
5
.0
0

2
0
.0
0

2
5
.0
0

3
0
.0
0

3
5
.0
0

4
0
.0
0

Retention
Indices

T
H
Y
M
A
S
T
F
.
D
\
F
I
D
1
A

T
im
e

1
0
0
0
0
0
0
9
0
0
0
0
0
8
0
0
0
0
0
7
0
0
0
0
0
6
0
0
0
0
0

Appropriated database

5
0
0
0
0
0
4
0
0
0
0
0
3
0
0
0
0
0
2
0
0
0
0
0

5
.
0
0

1
0
.
0
0

1
5
.
0
0

2
0
.
0
0

2
5
.
0
0

3
0
.
0
0

3
5
.
0
0

4
0
.
0
0

T
i
m
e

Identification
Unequivocal ?

A second identification methodology is required

Can GC help?

Components identification in complex volatile mixtures

Hyphenated methods

GC-FTIR

GC-MS
105

A
b
u
n
d
a
n
c
e

%T
100

S
c
a
n
3
4
8
8
(
1
5
.
1
4
2
m
in
)
:T
H
Y
M
U
S
A
.
D
1
3
5
4
5
0
0
0
0
0

4
0
0
0
0
0
0

95
3
5
0
0
0
0
0
1
5
0

90

3
0
0
0
0
0
0

2
5
0
0
0
0
0

85
2
0
0
0
0
0
0
9
1

80

1
5
0
0
0
0
0

1
1
5

1
0
0
0
0
0
0

601

741

882

1023

1164

1305

1445

1586

1727

1868

2008

2149

2290

2431

2572

2712

2853

2994

3135

3276

3416

cm-1

3557

75

7
7
5
0
0
0
0
0

5
3
95
1 6

1
0
3
2
0
7

0
4
0

6
0

8
0

1
0
0

1
2
0

1
4
0

1
6
0

1
8
0

2
0
0

2
2
0

2
6
7
2
8
1
2
4
0

2
6
0

2
8
0

m
/
z
>

A
b
u
n
d
a
n
c
e
T
I
C
:J
A
L
P
I
N
A
B
.
D

4
0
0
0
0
0

3
5
0
0
0
0

3
0
0
0
0
0

2
5
0
0
0
0

2
0
0
0
0
0

1
5
0
0
0
0

1
0
0
0
0
0

5
0
0
0
0

0
5
.
0
0
1
0
.
0
0
1
5
.
0
0
2
0
.
0
0
2
5
.
0
0
3
0
.
0
0
3
5
.
0
0
4
0
.
0
0
4
5
.
0
0
5
0
.
0
0
5
5
.
0
0
6
0
.
0
0
T
i
m
e
>

Components identification in complex volatile mixtures

Hyphenated methods : GC-MS
A
b
u
n
d
a
n
c
e
S
c
a
n
3
4
8
8
(
1
5
.
1
4
2
m
in
)
:T
H
Y
M
U
S
A
.
D
1
3
5

GC-MS

4
5
0
0
0
0
0

4
0
0
0
0
0
0

3
5
0
0
0
0
0
1
5
0
3
0
0
0
0
0
0

2
5
0
0
0
0
0

2
0
0
0
0
0
0
9
1
1
5
0
0
0
0
0

1
1
5

1
0
0
0
0
0
0
7
7
5
0
0
0
0
0

5
3
95
1 6

1
0
3
2
0
7

0
4
0

6
0

8
0

1
0
0

1
2
0

1
4
0

m
/
z
>

A
b
u
n
d
a
n
c
e
T
I
C
:J
A
L
P
I
N
A
B
.
D

4
0
0
0
0
0

3
5
0
0
0
0

3
0
0
0
0
0

2
5
0
0
0
0

2
0
0
0
0
0

1
5
0
0
0
0

1
0
0
0
0
0

5
0
0
0
0

0
5
.
0
0
1
0
.
0
0
1
5
.
0
0
2
0
.
0
0
2
5
.
0
0
3
0
.
0
0
3
5
.
0
0
4
0
.
0
0
4
5
.
0
0
5
0
.
0
0
5
5
.
0
0
6
0
.
0
0
T
i
m
e
>

1
6
0

1
8
0

2
0
0

2
2
0

2
6
7
2
8
1
2
4
0

2
6
0

2
8
0

Components identification in complex volatile mixtures

Hyphenated methods : GC-MS

Mass detectors
Quadrupole
Ion trap
Magnetic sector

Vacuum source

Turbomolecular pump
Diffusion pump

Components identification in complex volatile mixtures

Hyphenated methods : GC-MS
+

+
M/z=95

M/z=123

M/z=81

M/z=71

M/z=138
Abundance
Scan 2411 (11.112 min): BASI.D
71

30000

+
OH

95
81

25000

OH
M/z=156

20000

15000

123
55

10000

41

138

109

5000
65
0
30
m/z-->

40

50

60

87
70

80

90

154
100

110

120

130

140

150

160

170

Components identification in complex volatile mixtures

Hyphenated methods : GC-MS

Abundance
Scan 2411 (11.112 min): BASI.D
71

30000

95
81

25000

20000

15000

123
55

10000

41

138

109

5000
65
0
30
m/z-->

40

50

60

87
70

80

90

154
100

110

120

130

140

150

160

170

Components identification in complex volatile mixtures:

A combined dynamic methodology of 2x GC (RI) and GC-MS

Re
sp
o
n
se
_

2x GC (RI)

Quantification

T
HYM
AST
B.D\F
ID2
B

1
7
0
0
0
0
0
1
6
0
0
0
0
0
1
5
0
0
0
0
0
1
4
0
0
0
0
0
1
3
0
0
0
0
0
1
2
0
0
0
0
0
1
1
0
0
0
0
0
1
0
0
0
0
0
0
9
0
0
0
0
0
8
0
0
0
0
0
7
0
0
0
0
0

Retention
Indices

6
0
0
0
0
0
5
0
0
0
0
0
4
0
0
0
0
0
3
0
0
0
0
0
2
0
0
0
0
0

R
e
s
p
o
n
s
e
_

1
0
0
0
0
0
5
.0
0

1
0
.0
0

1
5
.0
0

2
0
.0
0

2
5
.0
0

3
0
.0
0

3
5
.0
0

4
0
.0
0

T
H
Y
M
A
S
T
F
.
D
\
F
I
D
1
A

T
im
e

1
0
0
0
0
0
0
9
0
0
0
0
0
8
0
0
0
0
0
7
0
0
0
0
0

Appropriated database

6
0
0
0
0
0
5
0
0
0
0
0
4
0
0
0
0
0
3
0
0
0
0
0
2
0
0
0
0
0

5
.
0
0

1
0
.
0
0

1
5
.
0
0

2
0
.
0
0

2
5
.
0
0

3
0
.
0
0

3
5
.
0
0

4
0
.
0
0

T
i
m
e

Identification

A
b
u
n
d
a
n
c
e

GC-MS

S
c
a
n
3
4
8
8
(
1
5
.
1
4
2
m
in
)
:T
H
Y
M
U
S
A
.
D
1
3
5
4
5
0
0
0
0
0

4
0
0
0
0
0
0

A
b
u
n
d
a
n
c
e

3
5
0
0
0
0
0
1
5
0

T
I
C
:T
H
Y
M
U
S
A
.
D

3
0
0
0
0
0
0

2
5
0
0
0
0
0

3
.
5
e
+
0
7

2
0
0
0
0
0
0
9
1

3
e
+
0
7
1
5
0
0
0
0
0

1
1
5

1
0
0
0
0
0
0

2
.
5
e
+
0
7

Mass spectra

7
7
5
0
0
0
0
0

5
3
95
1 6

1
0
3

2
e
+
0
7

2
0
7

0
4
0

6
0

8
0

1
0
0

1
2
0

1
4
0

1
6
0

1
8
0

2
0
0

2
2
0

2
6
7
2
8
1
2
4
0

2
6
0

2
8
0

m
/
z
>

1
.
5
e
+
0
7

1
e
+
0
7

5
0
0
0
0
0
0

0
5
.
0
0
T
im
e
>

1
0
.
0
0

1
5
.
0
0

2
0
.
0
0

2
5
.
0
0

3
0
.
0
0

3
5
.
0
0

4
0
.
0
0

4
5
.
0
0

Appropriated database