Analytical methods used in analysis of fragrance & flavour Materials

Analytical Methods Used
In analysis of Fragrance
& Flavour Materials
Presented By: Pravin S Mogarkar (18PER208)
Guide By: Dr. Satyajit Saha (ICT, Mumbai)
1

Contents
• Objectives
• Instruments used for Identification
• Instruments used for separation
• Instruments used for separation and identification
• Applications
2

Objectives
• Separation
• Identification
• Quantification
• Reverse Engineering
3

Instruments Used For Identification
IR
• Absorption of specific
frequencies by molecules
which is characteristic to
their structure.
MS
• Isolation of compound on
the basis of Mass/Charge
Ratio
4

NMR
• Structure prediction based
on nuclear spin.
E-NOSE
• Detection of volatile
organic compound.
5

Instruments Used For Separation
• Partition of volatile compound in
two phases.GC
• Difference in molecules affinity
to take longer to pass through
chromatography column.
HPLC
7

Gas Chromatography
8
• Mobile phase – Gas such as helium
• Stationary phase- liquid /solid
• Sample – gas/liquid
• Chromatograph temperature- 300℃

Working of Gas Chromatography
9

High Pressure Liquid Chromatography
10
• Mobile phase- liquid such as Water,
acetonitrile, methanol.
• Stationary phase- solid (silica)/liquid
• Type of stationary phase
1. Hydrophilic (Traditional HPLC)
2. Hydrophobic (RP- HPLC)

Instruments Used For Separation And
Identification
GC-IR GC-MS GC-O
11

FTIR
12
Sample used- solid or liquid
Frequency ranges- For functional group-
4000 cm−1 – 1300 cm−1
For Fingerprint region - 1300 cm−1 – 400 cm−1
(Molecule Fingerprints region )

GC-MS
13
Separation and identification based on
molecular mass.

Extraction Techniques
Extraction Techniques
Solid Phase Micro
Extraction
Microwave Assisted
Extraction
Super Critical Fluid
Extraction
Pressurized Fluid
Extraction
14

15
SOLID PHASE MICROEXTRACTION AND MICROWAVE
ASSISTED EXTRACTION

16
SUPER CRITICAL FLUID EXTRACTION

17
PRESSURIZED FLUID EXTRACTION

Characterization of the volatile substances and aroma components
from traditional soypaste.
• The study includes extraction of flavour substances in soypaste by simultaneous
distillation and their identification by GC-MS.
• Aroma compounds determined by GC-O & their FD value by AEDA method.
• It showed aroma of soypaste is due to presence of heterocyclic compounds and
organic acids.
19
Zhang et. al. (2010). Molecules, 15(5), 3421-3427.

Experimental Procedure
Fermentation
Sample Preparation For SDE Analysis
Analysis Of GC-MS Followed By GC-O
FD Value Analysis By Aroma Extract Dilution
Analysis Method
20

The contents of aldehydes and ketones is
lowest
21

The contents of acid and alcohol is highest
22Zhang et. al. (2010). Molecules, 15(5), 3421-3427.

Some ester components
23
Zhang et. al. (2010). Molecules, 15(5),
3421-3427.

Some heterocyclic components
24
Zhang et. al. (2010).
Molecules, 15(5), 3421-3427.

Analysis of the volatile components of five Turkish
Rhododendron species by headspace solid-phase
microextraction and GC-MS (HS-SPME-GC-MS).
• Volatile constituents of various solvent extracts of 15 different organs (leaves,
flowers, fruits) of five Rhododendron species (Ericaceae) growing in Turkey were
trapped with headspace solid-phase micro extraction (HS-SPME) technique .
• This Constituents then analyzed by GC-MS.
• The study indicated intra-specific variations in volatile compositions within the
genus.
25
Tasdemir et. al. (2003). Zeitschrift Für Naturforschung C, 58(11-12), 797-803.

26
Extraction
Headspace- Solid Phase Micro extraction
Analysis Of Volatile Compounds By GC-MS
Identification Of the Compound

27
Tasdemir et. al. (2003). Zeitschrift Für Naturforschung
C, 58(11-12), 797-803
Rhododendron species examined by HS-SPME coupled with
GC-MS

28
Tasdemir et. al. (2003). Zeitschrift Für Naturforschung
C, 58(11-12), 797-803
Volatiles identified from then-hexane extracts of Rhododendron species by
SPME-GC-MS

29
Tasdemir et. al. (2003). Zeitschrift Für
Naturforschung C, 58(11-12), 797-803
The volatiles identified from the CH2Cl2 extracts of Rhododendron by SPME-GC-MS

Fourier Transformed Infrared Spectroscopy Analysis of Constituents of Lemon
Essential Oils from Algeria
• The volatile compounds of Lemon (Citrus limon L.)is detected and
identified by Fourier Transformed Infrared Spectroscopy (FTIR) analysis.
• FTIR identify 10 volatile compounds and indicated than the functional
groups of the essential oils are C-H (Alkene), C-H (aromatic) and C=C.
• The obtained results have shown that the essential oils can be fully utilized
for pharmacy, cosmetology and industry.
30
Hicham Boughendjioua et.al. American Journal of Optics and
Photonics. Vol. 5, No. 3, 2017

Plant Materials
Isolation of the essential oil
FTIR analysis
31
Hicham Boughendjioua et.al. American Journal of
Optics and Photonics. Vol. 5, No. 3, 2017

32
Hicham Boughendjioua et.al. American Journal of
Optics and Photonics. Vol. 5, No. 3, 2017

GC-MS and FT-IR analysis of constituents of essential oil
from Cinnamon bark
• Cinnamon bark oil obtained by hydro distillation and further
analysis done by GC-MS and FT-IR to identify the component.
• Six components is identified and 2-propenal, 3-phenyl is the
major constituent (87.013%)
• Minor components Eugenol, O- Methoxy cinnamic aldehyde ,α-
muurolene , Naphthalene, 1,2,3,4-tetrahydro-1,6-dime, Tricyclo
nona-3,6-dien-9-on .
33
Bizuneh Adinew et.al. 2014, International
Journal of Herbal Medicine, 1(6), 22-31.

Plant material
Hydro distillation( Extraction)
Analysis of GC-MS
Analysis of FT-IR
34

35
GC-MS profile of
Cinnamon bark
essential oil

36
FT-IR spectra of
cinnamon bark
essential oil

Characterization of Aroma Active Compounds in Fruit Juice
and Peel Oil of Jinchen Sweet Orange Fruit (Citrus sinensis (L.)
Osbeck) by GC-MS and GC-O
• 49 and 32 compounds are identified in fruit juice and peel oil, respectively.
• GC-O is used to study the aromatic profile of Jinchen fruit juice and peel oil.
• 41 contribute to the aroma of fruit juice and peel oil.
• 12 is the odorants perceived in both samples.
• The aromatic compositions of fruit juice is more complex than that of peel oil.
• 19 components have been perceived only in the juice and 10 compounds is described as
aromatic components of only the peel oil by the panellists.
37
Yu Qiao et.al 2008 Molecules, 13(6), 1333-1344

Sample (Fruit Juice & peel oil)
Extraction by SPME
Analysis by GC-MS
Analysis by GC-O
38

39
Volatile components identified in fruit juice and peel oil of Jinchen fruit by
GC-MS.

40
Aroma active compounds of fruit juice and peel oil detected by GC-O

Applications and Advances in Electronic-Nose
Technologies
• In one of the research the author discussed a relationship
between the shape of the molecule, such as the presence of
certain functional groups, and features of certain olfactory
receptors that respond primarily to compounds with specific
three-dimensional molecular profiles elucidated in related
studies
41
Wilson et.al (20099(7), 5099-5148.

42
Examples of
primary aroma
categories proposed
by Amoore
Wilson et.al (20099(7), 5099-5148.

Electronic Noses’ and Their Application to Food
• At present, the commercial gas sensor technologies comprise metal oxide
semiconductors, metal oxide semiconductor field effect transistors, organic
conducting polymers, and piezoelectric crystal sensors.
43
Schaller et.al (1998), 31(4), 305-316.

44
Table- Application of
different 'electronic nose’
systems to different foods
Schaller et.al (1998), 31(4), 305-316.

Analytical methods used in analysis of fragrance & flavour Materials

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