分析化学会 GC研究懇談会

2017.12.1におい受容機構を考慮した
GC-MSデータの見方
埼玉大学大学院理工学研究科
長谷川 登志夫Saitama University
Graduate School of Science and Engineering
Toshio Hasegawa

題 目
１．におい受容機構を考慮した複合臭の取り扱いとは？

２．白檀などの香気素材の香気特性をどう取り扱うか？

３．実際のGCデータをどのように解釈したらいいのか？

2

におい受容機構
Recognitionof odorOdorant

Chemicalinteraction

Electricsignals
Olfactory receptor
Olfactory sensory neuron

ＡＢにおい分子をにおい分子受容体でとらえるにおい分子とにおい分子受容体相互作用の情報を脳で判断
3

Odor Receptors and Odor Molecules
Odor receptor（G-protein）

Odor molecule (ligand)
OHOCH3

Mouse mOR-EGCHOHOCHO

Mouse MOR23
HO

Human OR17-40

OO

Rat 17

CHO

4

Odor Receptors and Odor Molecules
Odor receptor（G-protein）

MousemOR-EG

Mouse
MOR23

Human
OR17-40

Rat 17

1:1

HOH

HO

OCH3

Odor molecule(ligand)

CHO

O

CHO

O

CHO

O

OH
OH

OCH3

OH
OCH2CH3

O
O

OCH3

CHO

Recognition

Non-recognition

Touhara K. et al., J. Neuroscience. 2005, 25, 1806-1815.
5

Odor Receptors and Odor Molecules
Mouse mOR-EG受容体
Vanilin
OH

OH

OH
OCH3

CHO

RecognitionOH
OH

OCH2CH3

OCH3

CHO

O
OCH3O

OCH3

Non-regognition

Touhara K. et al., J. Neuroscience. 2005, 25, 1806-1815.

6

Relationship Between Odorant and Receptorにおい分子とにおい受容体とは複雑な関係で相互作用している

におい分子

受容体
受容体

受容体

におい分子
受容体

におい分子

におい分子

におい分子受容体

受容体
受容体

１つのにおい分子は，異なった強度で複数のにおい分子受容体と相互作用する

におい分子

におい分子
におい分子１つのにおい分子受容体は，異なった強度で複数のにおい分子と相互作用する

7

Effect on Receptor when Odor Compounds with
Similar Structure are Mixed

The above literature indicates that the odor of a given material is not the simple
sum of the odors of each constituent.
And, the interactions of several constituents with similar structure areimportant for the aroma profile.
Oka, Y.; Omura, M.; Kataoka, H.; Touhara, K. The EMBO J. 2004, 23, 120-126.

8

Proposed Method: Aroma Profile Analysis
いくつかの匂いグループが合わさって香気素材香気を形成

におい分
子A-1

相互作用
におい分
子A-2

香気素材

におい分
子
B-1

におい分
子
B-2

相互作用
匂い分子
B-3

におい分
子A-3

scents
匂いグループA。。。。。。

匂いグループB

抽出
分別蒸留
香気成分抽出物
高沸点

低沸点＜抽出方法の違い＞＜沸点の違い＞9

題 目
１．におい受容機構を考慮した複合臭の取り扱いとは？

２．白檀などの香気素材の香気特性をどう取り扱うか？

３．実際のGCデータをどのように解釈したらいいのか？

10

Aroma ProfileofFrankincenseComparison of Constituentsof Hexane Extract and Steam-Distilled Oil
Hexane extraction
43％

Hexane extract（Relaxing odor ）Important odor constituentsOO

12OOHO

34OO

O

OOOO

5OO

DifferentFrankincense（Relaxing odor）

4％

H

Steam-distilled oil

Steam distillation（Pungent odor）

Octyl acetate
Octanol

1H NMR (200 MHz, CDCl )312

Investigation of Aroma Components of Frankincense

Hasegawa, T et al., . Natural Product Communications, 2012, 24, 593-598.
13

Main Constituentsin Hexane Extract of Frankincense
Diterpene hydrocarbonAliphatic alcohol and ester
22％

6 H(Verticilla-4(20), 7,11-triene)

OO

4％

Floral and fruity odor
OHO

Rose-like odor

2OH
(Incensole oxide)
1(Incensole)

2％O
No odorO

3
Weak fatty and fresh odor
O
(Incensole acetate)O
26％
Weak fatty andfresh odor

20％
Weak fatty and fresh odor
Diterpene alcohol and esters

16％

O
5O
O
(Incensoleoxideacetate B)

O
4O(Incensole oxideacetate A)
OO

10％Weak fatty and fresh odor

O

Small amount
Weak fatty and fresh odor

14Aroma Profile
ofSandalwood

Main Constituents of Sandalwood

OHOH
a-Santalolb-Santalol

Many studies have been reported on sandalwood, and the structure–odor relationships ofb-santalol and its related compounds have been investigated in detail .

[1] Stappen, I.; Hoefinghoff, J.; Friedl, S.; Pammer, C.; Wolschann, P.; Buchbauer, G., Eur. J. Med. Chem., 2008, 43, 1525-1529.
[2] Brocke, C.; Eh, M.; Finke, A. Chem. & Bio., 2008, 5, 1000-1010.
[3] Buchbauer, G.; Stappen, I.; Pretterklieber, C.; Wolschann, P., Eur. J. Med. Chem., 2004, 39, 1039-1046.
16

Investigation of Aroma Components of Sandalwood
Group AGroup BOils(Steam distilled)

OO

a-SantaleneO
H(Z)-a-Santalenyl
formateONew constituents

H(Z)-b-Santalenyl
formateCHO

CHO

Sandalwood
chips

Similar odor

b-Santalene(Z)-a-Santalal (E)-a-SantalalGroup C
0.2%

0.8%

99.0%

Residue
Hexane extract

Fractional distillationOH

a-SantalolOHb-Santalol

Hasegawa, T.; Toriyama, T.; Ohshima, N.; Tajima, Y.; Mimura, I.; Hirota, K.; Nagasaki, Y.,
and Yamada, H. Flavour and Fragrance Journal, 2011, 26, 98-100.

17

Aroma Profile of Sandalwood

白檀の香気プロフィール

Group AGroup BOOO

HO(Z)-a-santalenyl
formateHCHO(Z)-b-santalenyl
formatea-santalene(Z)-a-santalalCHOb-santalene(E)-a-santalalGroup COH

a-santalolOHb-santalol

18

Aroma ProfileofStar Anise

Constituents of Star Anise
Benzene derivatives(E)-Anethole
90.55%
Anise like odor

ρ-Anisaldehyde
1.45%

Estragole
0.67%(Z)-Anethole
0.13%

Methyl anisate0.07%

1-(4-Methoxyphenyl)-1-propanone
0.02%

Anisketone
0.12%

p-Cymene
0.11%

Terpenoid compounds

Limonene
3.3%

a-Pinene
0.7%

18 compounds, 6.8%

Benzene derivatives
93.2%

Methyl eugenol
0.12%

9 compounds, 93.2%
Szczepanik, M.; Szumny, A. Allelopathy J. 2011, 27, 277-288.

4 20

1H NMR Comparison of Components of Extracts

Obtained by Three Different Methods
1H NMR (CDCl , 500 MHz)3(E)-Anethole

MMSE method

Hexane extract

Steam distillation

p-Anisaldehyde(E)-Anethole (1):p-Anisaldehyde (2)
21:1

11:1

High

6:1

Low

Similarity of odor to star anise

21Aroma Profile of Star Anise
Monosubstituted benzene(E)-AnetholeOthers

p-Anisaldehyde

Para-disubstituted benzene

The odor of star anise consisted of the combination of
three groups of compounds.

22

Aroma ProfileofGreen Tea Leaves

23

Aroma Compounds in Green Tea Leaves
Greenish(3Z)-hexenol(2E)-hexenal(3E)-hexenol

FloralRoast
Dimethylsulfide

Geraniol

Linalool

2,5-Dimethylpyrazine

Green tea-like odor？

24

Reduction of Compounds with a Formyl group in the Residue
The residue

LiAlH4

Alcohols

Odor

Matcha-like

Matcha-likeComponents

Derived from
Formyl group

Derived from
Formyl group

1H NMR (500 MHz,CDCl )3

1H NMR (500 MHz,CDCl )325

Aroma Profile of Green Tea
Greenish(3Z)-hexenol

RoastFloral(2E)-hexenal(3E)-hexenol
Dimethylsulfide

Geraniol

Linalool

Matcha-like
2,5-Dimethylpyrazine(Z)-2-Nonenal

1-Octanal(E,Z)-2,6-Nonadienal(E,E)-2,4-Nonadienal

Safranal(E,Z)-2,4-Decadienal26

におい分子の構造の類似性とは？
種々のにおい分子構造変換におい分子同士が類似のにおいを示すにおい受容体は類似の構造として認識している
類似のにおい受容体グ
ループにおい分子同士が異なったにおいを示すにおい受容体は異なった構造として認識している
異なったにおい受容体グループ27

Aroma Profile of Sandalwood

白檀の香気プロフィール

Group AGroup BOOO

HO(Z)-a-santalenyl
formateHCHO(Z)-b-santalenyl
formatea-santalene(Z)-a-santalalCHOb-santalene(E)-a-santalalGroup COH

a-santalol

OHb-santalol

28

Structure-odor Relationship of a-Santalol

A
E

Saturated

BZ

C
a-Santalol

Part A: Geometric isomers
Part B: Replacement of functional groups

Dramaticodor changePart C: Loss of bulky polycyclic moiety
T. Hasegawa,; H. Izumi,; H. Yamada.; Natural Product Communications, 2013, 8, 1-3.

29

Effect of changing part A on odor
E-Isomer

Z-IsomerWoodySandalwood-like,sweetOdorless

OOO
O

Different
odor

CHOOHOH

Slightly fatty
OOHO

Slightly fresh

Odorless

CH3

CH3Fatty, woody

Green woody

CHO

OH

Characteristic woody odor

Very weak odor

Saturated compound

Similar odor

OHA
C

Sweet, woody

Sandalwood-like,fresh, green

OOO

OH

B

Different odor

CHO

H

Floral, woody

O

Woody odors

CH3Fatty, woody

Hasegawa, T. et al., Molecules. 2012, 17, 2259-2270.

30Effect of changing part B on odor
Z-Isomer

Saturated compound

E-Isomer

ZE

Differentodor

Medicinal,
fatty

Introduction of benzyl group

Sweet

Similar
odor

Sweet

Dramatic change in odor,
loss of woody odor

ABC

Similarodor

Different
odor

X＝31

Effect of part C on odor

ACOH Bn-Butyl group

Linear alkyl group

Bulky polycyclic moiety
32Effect of changing part C on odor
Z-Isomer

Saturated compound

E-IsomerC
Woody

Similarodor

Medicinal, fatty

Different
odor

Sweet woody

Different
odor

odorless

Sweet

Similar
odor

Sweet

Similarodor

Fatty, floral
slightly sweetC
Fatty, floral,slightly sweet

Fresh, fatty

Similarodor

Fatty, floral

Fresh, fatty

Fresh, fatty

Loss of woody note and difference in odors between geometric isomers
33α-サンタロールの構造と匂いの関係
Structure-Odor Relationship of a-Santalol

A

Hydroxy

FormylOH

n-Butyl groupC

α-Santalol

B

Benzyl group
Formyloxy

Acetoxy

Part A: Geometric isomers
Part B: Replacement of functional groups

Dramatic
odor changePart C: Loss of bulky polycyclic moiety
T. Hasegawa,; H. Izumi,; H. Yamada.; Natural Product Communications, 2013, 8, 1-3.

34

Aroma Profile of Star Anise
Monosubstituted benzene(E)-Anethole

Othersp-Anisaldehyde

Para-disubstituted benzene

The odor of star anise consisted of the combination of
three groups of compounds.

35

Structural Features That Affect the Odor ofAnetholeAnetholeB

Double bondIn the side chainCPolar group

Santalol1)

System 1
Sour
fresh
fruityAniselike

fresh
Fatty

Unpleasant

Slightly
Sweet

Woody

Fatty
fruityAniselike

Fatty
fresh
floralFatty
fruitySlightly
sweet
fatty

Woody

Fatty
fresh
sweet

Slightly
fatty
fresh

Fatty
floralSystem 2

AMethoxy group

System 3
Fatty
floral

Fatty
sweet

Fatty

Removing the p-substituted benzene moiety caused
the loss of characteristic odors.
361) Hasegawa, T.; Izumi, H.; Yamada, H. Natural Product Communications, 2013, 8, 1-3.

複合臭の香気特性の解明

臭い受容機構に基づいたアプローチ
有機化学的観点による受容機構の解明

香気プロフィール解析

構造と香りの関係

37

題 目
１．におい受容機構を考慮した複合臭の取り扱いとは？

２．白檀などの香気素材の香気特性をどう取り扱うか？

３．実際のGCデータをどのように解釈したらいいのか？

38

Aroma ProfileofGalangal

39

Studies of the Components and Aroma of Galangal
Borneol(1.2%)
1,8-Cineole(2.4%)

Pentadecane(4.9%)(E)-Ethyl 4-methoxycinnamate(52.5%)

Ethylcinnamate(26.3%)“The cinnamate derivatives are responsible for the aromatic-spicy odor impression,
whereas especially the monoterpenes, like 1,8-cineole, borneol, d-3-carene, carvone
and carvone oxide generally possess pleasant-fresh odor notes.”
Leopold Jirovetz et al. Acta Pharmaceutica Turcica, 2001, 43, 107–110.

Galangal contains various aroma components.
However, it is not clear which compounds contribute to its aroma.
4040

Aroma Profile of Star Anise
The ratio of the two main components
Aroma profile of star anise(E)-Anethole:p-anisaldehyde(E)-Anetholep-AnisaldehydeMMSE *21 : 1

Hexane extract

11 : 1

Steam distillate

6:1*Monolithic Material Sorptive Extractiona)Odorsimilarity
to the materialhighlowa) Calculated by 1H NMR

The ratio of (E)-anethole to p-anisaldehyde was associated withthe level of odor similarity to star anise.
Hasegawa, T. ; Seimiya, H. ; Fujihara, T. ; Fujiwara, N. ; Yamada, H. Natural Product Communications, 2014, 9, 251–256.

＋
Structurally
similarcompounds?

Does the ratio of these cinnamic acid derivatives
affect the odor of galangal...?
41Investigation of Galangal Odor by Approach I
Pentadecane,odorless

Hexane extract

Extractability 3.67%,
room temperature

60 °C

Ethyl cinnamate (E)-Ethyl 4-methoxycinnamate

1：

10Spicy, fresh, sour

Headspace sampling(MMSE extract)

Odor similarity
to galangal
High1：

31：

7Spicy, fresh, sour

Extractability 1.68%
Steam distillate

Low
Fresh, medicinal

The aroma didn’t depend on the ratio of the cinnamic acid derivatives.
42

Searching for Key Odor Compounds by Approach II
Hexane extract

Galangal-like odor
Low bp20–62 °C
0.20–0.60 Torr

Fraction 1Spicy, sour

12%*Ethylcinnamate
62–84 °C
0.20–0.80 Torr

Fraction 2
Spicy, sour, sweet77%*
Ethyl 4-methoxycinnamate

84–175 °C
0.20–0.70 TorrHigh bp

Fraction 3* Percentage in the hexane extract

Sour, medicinal

ResidueSlightly sour

Bulb-to-bulb distillation

Ethyl cinnamate made a larger
contribution to the aroma ofgalangal than(E)-ethyl 4-methoxycinnamate.

43

GC-MS Analysis of the Components in Fraction 2⑤⑩④①②5③⑱⑨⑪
⑥⑦⑧

⑭⑫ ⑬⑮

10⑳⑯⑰

15⑲202530
minutes① Tridecane② Tetradecane③ a-Ylangene④ a-Copaene⑤ Pentadecane⑥ a-Gurjunene⑦ Humulene⑧ endo-Borneol②Linear chain compounds③⑨ Heptadecane⑩ a-Selinene⑪ 8-Heptadecene⑫ g-Muurolene⑬ 2-Tridecanone⑭ Anethole⑮ p-Cymen-8-ol⑯ (Z)-Ethyl cinnamate⑫⑭

Cyclic terpenes⑰ Anisaldehyde⑱ (E)-Ethyl cinnamate⑲ (Z)-Ethyl 4-methoxycinnamate⑳ (E)-Ethyl 4-methoxycinnamate

Aromatic compounds

etc.
Cyclic terpenes

44

含有成分の構造類似性を考慮したGC-MSデータ解析⑤⑩④①②③5⑱⑨⑪⑥⑦⑧⑭⑳⑫ ⑬⑮10⑯⑰

15⑲202530minutes① Tridecane② Tetradecane③ a-Ylangene④ a-Copaene⑤ Pentadecane⑥ a-Gurjunene⑦ Humulene⑧ endo-Borneol⑨ Heptadecane⑩ a-Selinene⑪ 8-Heptadecene⑫ g-Muurolene

Aromatic compounds⑭⑬ 2-Tridecanone⑭ Anethole⑮ p-Cymen-8-ol⑯ (Z)-Ethyl cinnamate⑰ Anisaldehyde⑱ (E)-Ethyl cinnamate⑲ (Z)-Ethyl 4-methoxycinnamate⑳ (E)-Ethyl 4-methoxycinnamate

Cyclic terpenes⑱③Linear chain compounds⑩②⑤⑰⑳⑧etc.

⑨⑫etc.

⑪These compounds were classified as aromatic compounds, cyclic terpenes,
and linear chain compounds, according to their characteristic structures.

etc.
45