GCMS presentation - natural products

The Metabolite Map for Identifying

Probabilities and Olfactory Analysis As A Supplement to GCMS

Analysis Murray Hunter

Oxygen

Carbon Dioxide

Water

Nitrates & Mineral Salts

Glycose

Polysaccharides & Disaccharides

Glycolysis

6-Deoxyxylulose

Isoprenoids (terpenoids)

Erythrose 4-phosphate

Pentose Phosphate Pathway

Phosphoenol pyruate

Pyruvate Hydroxy-

Benzoic Acid

Shikimate Pathway

Alkaloids

Phenylpropanoids

Complex Alkaloids

KREBS CYCLE

Acetyl-CoA

Malonyl-CoA

Mevalonic Acid

Complex Isoprenoids (terpenoids) Polyketides

Flavonoids

Complex Flavonoids

Primary Metabolites (in blue) Secondary Metabolites (in red)

Aromatic Amino Acids

Amino Acids & Nucleotides

Photosynthesis

Triglycerides

2. Using CGMS in Forensic Fragrance Construction

3. Using CGMS in Identifying Essential oil constituents

4. Using CGMS in Quality Assurance of Flavours & fragrances

1. Olfactory Evaluation

What are essential oils?

Essential OilA volatile oil obtained from a wide variety of plant, scrub, and tree species and from various parts of the plant anatomy, such as the roots, rhizomes, wood bark, leaves, stems, fruit, flowers and seeds. Usually extracted by hydro or steam distillation, expression or effleurage - Hunter 1996

Concretes and Absolutes

Volatiles and waxes extracted from plant material with hydrocarbon solvents (usually benzene and hexane) through washing and removal of the volatile solvent with distillation. A waxy aromatic substance remaining is called a concrete. The concrete is washed with alcohol to remove the volatile materials and ethanol removed through vacuum distillation to leave an absolute.

Plant Material

Expressed

Oil

Cold expression of citrus fruits

Terpeneless Oil Terpene Tails

Essential

Oil

Water, water & steam and steam

Distillation

Absolute

Concrete

Solvent Extraction

Washing with ethanol and vacuum distilling

ethanol away

Pomade

Enfleurage

Vacuum distill away the ethanol

Oleoresin

CO2

Extract

CO2 Extraction

The Natural Aromatic Product Family

Olfactory Evaluation

?

The Olfactory System & Odour Classification

The Human Olfactory System

Odour molecules pass through the olfactory organ

Olfactory epithelium

• Olfactory mucosa– Mucus!– High in nasal cavity– Site of transduction– Contains olfactory

receptor neurons (ORN)

11

Nose hair: Olfactory cilia• ORN have cilia• Cilia contain olfactory

receptor proteins– Similar to visual

pigment• Transduction

– Odorants bind to ORs– Change shape of

protein– Ion flow across OR– Electricity

TransmissionOlfactory ReceptorsOlfactory Cells line the Olfactory Epithelium which is responsible for olfactionEach cell has cilia where receptor sites are locatedReplaced every monthAxons of the olfactory receptors carry information to the olfactory bulbOlfactory bulb sends axons to several specific parts of the cortex with precise connections From the cortex, information is sent to other areas that control feeding & reproduction

Olfactory DisorderAnosmia: the general lack of olfactionSpecific Anosmia: the inability to smell a specific chemical

13

How many receptor types are there?

• 1000 different kinds of olfactory receptors (OR)• 10 million OR neurons

– 10,000 of each type of OR– Each OR neuron has only one type of receptor– 1000 neuronal chemical detectors– Potential to differentiate between 5,000-10,000 different

odours

Olfactory Bulbs

• An outcropping of the brain

• Its like a snail in your brain!

• Electrical responses in cilia passed through olfactory nerve to OB

The hedonic primacy of olfaction• Sensory and emotional experience• Not the same for vision/audition

– Seeing and feeling more distinct• More intertwined in the chemical senses

– Why?• Orbitofrontal cortex

– Plays multiple roles– Critical for emotional experience– Secondary sensory cortex for olfaction– Also centre of creativity and imagination

The olfactory interpretation process from input to response

Nobel Prize

• October 4, 2004 - Richard Axel and Linda Buck honored with the 2004 Nobel Prize in Physiology or Medicine for pioneering studies that clarify how the olfactory system works.

Top Notes

Middle Notes

Base Notes

The Structure of a Fragrance

Bergamot oil, Rosewood oil (linalool), Linalyl acetate, Neroli oil, Ciste Oil

Rose Oil 5%, Jasmin absolute 4%, Ylang Ylang Oil, Aldehyde C11, C12, Methyl ionone 8%, hydroxycitronellal 10%, Cinamic Alcohol, Styrax. Phenol ethyl alcohol, Phenylacetaldehyde

Vetiveryl acetate, Sandalwood, Isoeugenol, Vanillin 1.5%, Coumarin 15%, Nitromusks 10%

“First Impression” in Perfume. High Volatile Citrus, Fresh, Green notes…..

“ Heart of fragrance”. Intermediate VolaleFloral, Aldehydic notes…..

Basic of fragrance”.(Bottom) Low VolatileWoody, Powdery, Musky …..

Classification of Odours

Spicy

Medicated

Aldehydic

Marine

Coniferous

Minty

Green

Herb -aceous

Fruity

Balsamic

Animalic

Woody

Floral

Agrestic

Curtis & Williams (1994)

Floral Family Broom Sweet, Floral, Herbaecious Carnation Delicate, Floral, CloveCassie Floral, Orange flower, VioletClover Sweet, Honey, Herbaecious, FloralCyclamen Floral, Lily, Lilac, Violet, greenGardenia Floral, Tuberose, Jasmin, Orange flowr, GreenHawthorn Floral, Bitter almond, Bitter, DiffusiveHeliotrope Floral, Almond, Balsamic, FruityHoneysuckle Swet, Floral, Heavy, Orange Flower, Tuberose,

Honey, RoseHyacinth Fresh, Green, Floral, Jasmin, BalsamicJ asmin Sweet, Floral, Heavy, Fruity, HerbaeciousJ onquil Fresh, Floral, Sweet, Heavy, Honey, GreenLilac Fresh, Meadow-Floral, Jasmin, GreenLily Sweet, Heavy, FloralLily of the Valley Floral, Rose, Green, Slightly citrusLime Blossom Floral, Fresh, Lily, Lilac, Orange FlowerMagnolia Floral, Lily, Ylang YlangMimosa Sweet, Floral, Hawthorn, Orange flower, GreenNarcissus Delicate, Fresh, Green, Floral, SweetOrange Flower Floral, Heavy, Animalic, Fresh, Rich, BitterOrris Delicate, Dry, Woody, Somewhat VioletReseda Floral, Green, Anisic, HerbaceousRose Sweet, Floral, Honey, Waxy, Slightly spicy,

FruitySweet Pea Sweet, Delicte, Floral, Orange Flower, Hyacinth,

LilyTuberose Heavy, Floral, Orange Flower, Ylang Ylang,

Caramel, GreenViolet Floral, Powdery, GreenWallflower Floral, somewhat liliacy, bitter almondYlang Ylang Rich, Sweet, Medicated, Floral, Fruity, Clove

Floral Family Descriptions

Woody Family Cedarwood Woody and slightly earthy, with smoky notesHibawood Like cedarwood, but more intensely woody,

Dry, PungentRosewood Woody, Floral, Sweet, Spicy, Delicately FattySandalwood Soft, Sweet, Woody, Slightly Balsamic

Woody Family Descriptions

Animalic Family Amine Fishy notes, AmmoniaCastoreum Warm, Phenolic, Sweet, Somewhat herbaceous,

clean smellingCatty Feline odourCivet Warm, Slightly faecal, MuskyEquine Horse likeFaecal Indolic, skatolicIndolic Lilac, Heavy, Animalic, NaphthalenicMusky Blackberry likeLeather Phenolic, Cresylic, Animalic, Castoreum,

Balsamic

Animalic Family Descriptions

Balsamic Family Cistus Rich, Balsamic, Ambergris, HerbaceousLabdanum Rich, Balsamic, AmbergrisMyrrh Rich, Warm, Balsamic, SpicyOpopanax Warm, Vegetable note, Rich, Balsamic, SpicyPeru Balsam Sweet, Balsamic, Rich, Soft, Vanilla,

Cinnamate, BenzoateStyrax Sweet, Balsamic, CinnamateTolu Balsam Sweet, Balsamic, Cinnamate, VanillaVanilla Soft, Sweet, Powdery, Balsamic

Balsamic Family Descriptions

Herbaceous Family Lavender Fruity, Floral, Herbaceous, Woody, BalsamicRosemary Herbaceous, Resinous, Woody, BalsamicSage Herbaceous, Slightly camphoraceous

Herbaceous Family Descriptions

Agrestic Family Earthy Fresh, Woody, VegetableForest Moist, Fresh, Vegatable, WoodyFungal Mushroom likeGalbanum Sharp, Agrestic, Green, Earthy, ConiferousHay Sweet, Warm, Agrestic, Herbaceous

Agrestic Family Descriptions

Green Family Cress Hyacinth like, GreenCucumber Green like sliced green vegetable pods

Fresh and CleanGrassy Fresh, Green, FruityLeafy Light, Green

Green Family Descriptions

Minty Family Peppermint Fresh, Cool, Mentholitic, Minty, Slightly GreenSpearmint Sweet, Warm, Minty, Herbaceous, GreenMossy Notes Forest, Woody, Phenolic, Green, Somewhat

marine

Minty Family Descriptions

Coniferous Family Description

Coniferous Family Pine Resinous, Balsamic, TerpeniticResinous Forest, Woody, Terpenite, Balsamic

Marine Family Ambergris Marine, Seaweed, Mossy, DryBeachOzonic Fresh, Marine, Slightly SweetSeaweed Marine, Mossy, Amine

Marine Family Descriptions

Aldehydic Family Fatty, Waxy (pleasant in dilution)

Aldehydic Family Descriptions

Medicated Family Camphorous CamphorCineolic Eucalyptus, Cresylic Cresolic, PhenolicEthereol alcoholicMentholic MentholPhenolic PhenolTerpenic monoterpenesThymolic ThymolWintergreen Methyl salicylate

Medicated Family Descriptions

Fruity Family Bergamot Fresh, Lemon, Sweet Orange, Herbaceous,(Citrus Sub-Class) Pepper, Floral

GrapfruitLemonLimeMandarinOrange SweetOrange BitterPithy Orange PeelTangerine

Fruity Family AppleApricotBananaBlackberryBlackcurrentPearPeachPineapplePruneRasberryStrawberryVinous Cognac likeWatermellon

Fruity Family Descriptions

Sub-Class (Citrus)

Spicy Family CeleryCinnamomCloveCorianderCuminFenugreekGingerNutmegPepper

Spicy Family Description

Miscellanious Bitter AlmondNotes Anisic

BenzoateBurntCaramelCinnamateCoconutFattyGeraniumHoneyMetallicNaphthenicNondescriptOilyPatchouliSalicylateSmokySulphorousTobaccoWaxy

Miscellaneous Notes

Light HeavySweet DryFresh StaleWarm CoolSoft Hard

Smooth HarshRich Thin

Delicate Coarse

Odour Characteristics

BitterDifussivePowderyPungentSharp

Tropical

Additional Characteristics

Spicy

Medicated

Aldehydic

Marine

Coniferous

Minty Green

Herb aceous

Fruity

Balsamic

Animalic

Woody

Floral

Agrestic

Fragrance Description Workshop

0 Bitter 10 0 Diffusive 10 0 Powdery 10 0 Pungent 10 0 Sharp 10 0 Tropical 10

GC-MS

Performance Test

A Fragrance Development Lab

A trained human more accurate than a GC-MS

Sample of a Flavour Chart

Perfumer Excellence

Knowledge (vocabulary) of

odorous substances

Knowledge of potential strengths,

weaknesses and applications of

odorous materials

Knowledge of outstanding

fragrance creations within the domain

Practical knowledge & experience

Olfactory sensitivity

Time, patience, perseverance

Interest and passion

Imagination

Knowledge Base Emotion

Cognitive Skill Creativity Tool

Process & Product

Curiosity, enquiry and

experimentation

Using CGMS in Forensic

Fragrance Construction

From This

To This

Apples aren't the same

Sweet, sour, green, red, sharp, fresh, wholesome, etc.

This means different aromatic chemicals play a role in fragrance

Discovered in 1937, the First Publication on Raspberry Ketone appears in 1961

O

HO

1964 – Introduction of Furaneol®

1965 – First publications on isolation of Furaneol®from strawberry and pineapple

O

O OH

O

O OH

1965-66 – α- & β- Sinensal isolated from orange oil

CHO

CHO

1960’s – Nootkatone

First isolated in 1962, it’s importance to citrus flavor was unknown until it’s isolation from grapefruit by Mcleod in 1964.In 1966, Coca-Cola introduced Fresca. By 1970, Nootkatone was GRAS and became a key ingredient in Fresca. Coca-Cola manufactured Nootkatone for many years and soon made it available to the flavor industry.

OO

(+)-Nootkatonestrong grapefruit odor, bitter in taste

(-)-Nootkatoneweak woody (vetiver note); no grapefruit character; virtually no taste

Threshold: 60,000 ppb 800 ppb

1970’s-80’s – Other Furanones

Sotolon (caramel furanone) & Maple furanone identified in cane sugar – powerful caramel maple notes

O

HO

OO

HO

O

Sotolon Maple Furanone (Abhexone)

Threshold: 0.003 ppb 0.00005 ppb

Sotolon – key flavorant of fenugreek, sake, sherry wine

1968 – Pfizer patents Ethyl maltol

O

OH

O

Ethyl maltol (Veltol Plus®) was touted to be about 6Xstronger than maltol and an important substitute for Coumarin. Well, at least it is stronger than maltol.

1975 - Introduction of α-Damascone and β-Damascone1982 – Introduction of Damascenone

1970 – Discovery Damascenone & β-Damascone

OO O

1974 – Thiomenthones identified in Buchu oil

OHS

OHS

Key component for black currant and the “fuzzy” peach skin note

1980 – Patent on Oxane®

1977 - Winter identifies 2-methyl-4-propyl-1,3-oxathiane as a character impact compound in Passionfruit 1984 – Pickenhagen determines that (-)- cis- 2-methyl-4-propyl-1,3-oxathiane is the important diastereomer

O

S

H H

(-)-(2R,4S)-2-methyl-4-propyl-1,3-oxathiane

• Odor: powerful sulfury, tropical fruit• Threshold: 2 ppb

O

S

HH

(+)-(2S,4R)-2-methyl-4-propyl-1,3-oxathiane

• Odor: flat, estery, camphoraceous, floral, less sulfury • Threshold:4 ppb

SH

S

1982 – 1-p-Menthene-8-thiol

Demole identifies 1-p-Menthene-8-thiol as a character impact compound in Grapefruit Juice

Very powerful with an odor threshold of 0.0001 ppb, it alsorequires stabilization as it tends to rapidly cyclize to the thio analog of dihydropinol.

O

O

O

O

O

O

Mintlactone (1983) Dihydromintlactone (1995)Wine lactone (1996)

GRAS No. -- 3764 4032Odor: Vinous, lactonic Minty, coumarinic, Coumarinic, hay,

(coconut) tenacious lactonic, tonkaFlavor: Lactonic, sweet, herbal Coumarinic, fatty, Coumarinic,

herbal lactonic, coconutThreshold*: 0.0000016ppb 0.00012 ppb 0.000039 ppb*In airRef: Frerot, et. al., Flavour Fragr. J., 2002; 17: 218–226; Gaudin, Tetrahedron, 2000, 56: 4769-4776

Coumarinic p-Menthane Lactones

1962 – First publications appear on Methyl Jasmonate & Methyl Dihydrojasmonate

1R,2R-(-)-Z-methyl jasmonate 1S,2S-(+)-Z-methyl jasmonate

4

5

1(R)

2 (R)

3

O

(Z)

CO2CH3

H

H

(S)(S)

O

(Z)

H3CO2C

H

H

(R)(S)

O

(Z)

CO2CH3

H

H

(S)(R)

O

(Z)

H3CO2C

H

H

1R,2S-(+)-Z-methyl epijasmonate 1S,2R-(-)-Z-methyl epijasmonate

Methyl Jasmonate – odorant, insect pheromone, plant growth regulator

Advances in Flavoring Materials

Chirality & Odor - Methyl Jasmonate & Methyl DihydrojasmonateConfiguration Odor Description Odor Threshold (in PPB)Methyl jasmonates1R,2R-(-)- Weak odor >70*1S,2S-(+)- Odorless Odorless1S,2R-(-)-epi- Odorless Odorless1R,2S-(+)-epi- Strong odor; floral, true jasmin-like 3*Ref: T.E. Acree et. al., J.Agric.Food.Chem. 1985Methyl dihydrojasmonates1R,2R-(-)- Floral, sweet, jasminelike 240**1S,2S-(+)- Floral, fatty, cis-jasmone, hay character, 15,360**

tea note, slightly lemon peellike (weak)1S,2R-(-)-epi- Herbal, fatty, tea-like, tobacco, 12,500**

β-damascone, cis-jasmone1R,2S-(+)-epi- Intensely floral, jasmine-like, bright, cis- 15**

jasmone, slightly fatty, woody, β-ionone-like, extremely long lasting

Ref: Werkhoff, P., et. al., Food Reviews International, 2002* detection threshold**recognition threshold

Advances in Flavoring Materials

2001 – Takasago’s Professor Ryoji Noyori wins the 2001 Nobel Prize for Chemistry

HOOH

Probable Constituents of an essential oil

Individual Aromatic Chemicals

12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.000

2000000

4000000

6000000

8000000

1e+07

1.2e+07

1.4e+07

1.6e+07

Time-->

•Abundance

GC-MS Analysis of a Meat Flavor

Peak Identified as Furfuryl MercaptanPowerful Coffee Aroma

Peaks are 2-Methyl-2,3-dihydrofuran-3-thiol isomersPowerful meat-like aroma

O

SH

CH3 O

SH

CH3

O

SH

• Citrus

• Floral

• Aldehydic

• Spicy

• Oriental

• Chypre

• Fougere

Basic Fragrance Types

Representative IngredientsNatural : Lemon Oil, Bergamote, Lime….Chemical : Citral, Dihydo Myrcenol….

Representative Fine Fragrance

O de Lancome (1975. Lancome)Bulgari Eau Parfume (1992. Bulgari)CK one (1994. C.Klein)

Eau Savage (1966.C.Dior)

BergamotLemonOrange

Rose JasmineLily of Valley

OakmossAmbergrisCivet

• Citrus notes Citrus + Floral

• Citrus notes

BergamotLemonMandarinDihydro Myrcenol

JasminMuguet

AmberMuskSandalwood

Green

Citrus

Floral

Amber

Woody Musk

• Floral notes Floral + Floral

Representative IngredientsNatural : Jamine Abs, Ylang Ylang, Rose Abs. Tuberose ….Chemical : Hedion, Benzyl Acetate….


Joy (1935. Jean Patou)Diorssimo (1956.C.Dior)Anais Anais (1979. Cacharel)Paris (1983. YSL)Beautiful (1985. E. Lauder)

Green

JasmineRose Ylang YlangVioletMuguet

MuskSandalwoodPowdery

Green

Floral

MuskWoodyPowdery

• Aldehydic notes Floral + Aldehydic

Aldehyd 10MuguetYlang-Ylang

JasmineRoseCarnationAldehyde C-11Aldehyde C-12

VanillaSandalwoodMusk

Floral

Aldehyddic

WoodyPowder

Representative IngredientsNatural : Not available Chemical : Aldehyde C-11, Aldehyde C-12..….


Chanel No. 5 (1921. Chanel)Calandre (1969. 1969. Rabanne)First (1976. Van Cleef & Arpels)

• Spicy notes Floral + Spicy

BergamotYlang Ylang

CarnationJasmineRoseIris

SandalwoodCedarwoodMusk

Floral

Spicy Floral

WoodyMusk

Representative IngredientsNatural : Clove Buds oil, Pepper oil, Cinnamon oil. Carnation……. Chemical : Eugenol, Cinnamic Aldehyde ..….


L’air du Temps (1948. Nina Ricci)Fidji (1966. Guy Laroche)

Egoist (1990. Chanel)

• Chypre notes What is Chypre….?

Citrus Green Aldehyde

Floral (Jasmine, Rose, Ylang…)

WoodyMossyPatchouli


Mitsouko (1919. Guerlain)Miss Dior (1947. Christian Dior)Coco (1984. Chanel)Ysatis (1984. Givenchy)

Representative IngredientsNatural : Oak moss Abs. Vertiver, Patchouli oil Galbanum Cederwood, SandalwoodChemical : Veramoss, Iso E Super..….

BergamotMandarinGalbanumAldehydC-11

JasmineRose Ylang-YlangMuguet

AmberMossyPatchouliMossyWoody

Oriental

CitrusGreenAldehyde

Floral

AmberWoodyMossy

• Chypre notes - Fine Fragrance

• Oriental notes What is Oriental….?

Citrus MandarinLemon

Spicy(cinamon..)Floral

Vanilla Abs.SweetBalsam

Representative IngredientsNatural : Vanilla, Tolu Balsam, Mandarin, Cinnamon Chemical : Vanillin, Galaxolide..….


Shalimar (1925. Guelain)Obsession (1984. Calvin Klein)Opium (1977. YSL)Samsara (1989. Guelain)Jean Paul Gaultier (1993. J.P.Gaultier)

BergamotLavendinCloveSpearmint

MuguetGeraniumCarnation

PowderyMossyCederwood

Herbacious

Floral

WoodyPowdery

• Fougere notes - Fine Fragrance

Using CGMS in Identifying Essential oil constituents

Biological Screening

Bio-prospecting Literature Review

Consider Crop Reintroduction Consider New Crop Introduction

Identify Chemical Constituents of Essential Oil

Screening for Development Potential

o Match chemical constituents of essential oil

with possible market uses o Evaluate application potential of essential oil o Evaluate theoretical yields, cost of production o Evaluate time and cost of development

Value Determination (Use Criteria) Preliminary study due to volume of possibilities

The Screening Process

Regulatory Screening

(FDAs, REACH, SCCP, BPD, etc)

Generation of Possibilities

Elimination of Possibilities

Leads to a number of potential possibilities that can be further studied

Potential essential oil crop

idea

Bio-prospecting Screening Protocols o Anti inflammatory o Anti microbial o UV absorbing o Anti ageing actives o Flavour & fragrance

application o Aromatherapy o Anti cancer

Desktop study

Knowledge Required o Essential oil applications o International market

(Flavour & fragrance, cosmetics, personal care, agro-chemical, aromatherapy, etc.)

o International regulations

Need to collaborate with industry parties

Ethno-botany

Other literature (Journals,

etc.)

Study of same

latitude Crops

What has value to industry?

o Chemical

constituents o Odour/flavour

profile o Potential

applications

Match chemical constituents with possible market

uses

Evaluate application potential of essential oil

Evaluate theoretical yields, cost of production

Evaluate time and cost of

development

Value Determination (Preliminary study due to volume of possibilities)

Choice and Access to Market

Jurisdiction Regulatory Framework

Ylang Ylang (Cananga odorata)

benzyl acetate (ca. 25 %), p-cresyl methyl ether (ca. 20 %), methyl benzoate (ca. 5 %), methyl salicylate, cinnamyl acetate, (-)-linalool (ca. 15 %), geranyl acetate (ca. 10 %), farnesyl acetate (ca. 3 %), as well as a number of other sesquiterpenes and their oxygenated derivatives, e.g. muurolol T (ca. 2 %)

Extraction Method: Steam distillation of the flowers

Origin: Indonesia, Madagascar

Patchouli Oil (Pogostemon cablin)

(-)-patchoulol andnorpatchoulenol

Obtained by steam distillation under pressure or CO2-extraction of the dried leaves

Patchouli is mostly grown in Indonesia

There are no synthetic equivalents of the patchouli scent.

Main Constituents: (-)-patchoulol (30-40 %). However, it is maintained that norpatchoulenol, present in only 0.3-0.4 %, is playing a principal part in the overall odour picture.

Pandanus Oil (Pandanus odoratissimus)

phenethyl methyl etherpandanol

Origin: native of South East Asia and is much cultivated on the Indian East Coast

The flowers are hydro-distilled to yield a 'kewda attar

phenethyl methyl ether (pandanol) (38 %), together with terpinen-4-ol (19 %), alpha-terpineol (8 %) and phenethyl alcohol (7 %) [79]. Phenethyl alcohol and its derivatives are common odorants in flowers

Backhousia citriodora (Lemon Myrtle)

High Investment to Expand Industry in Australia

Strong Demand as an Ingredient for tea

Good Crop to Grow in Most Parts of Thailand

CHO

CHO

Citral (geranial 51.43 % and neral 42.12 % )

Important New Compounds Rose oil – What’s Important

Component % of OilThreshold in

ppbOdor Units

x 10-3

Rel. % of odor

units(-)-Citronellol 38 40 9500 4.3C14 - C16 Paraffins 16 - - -Geraniol 14 75 1860 0.8Nerol 7 300 233 0.1Phenethyl alcohol 2.8 750 37 0.016Eugenol methyl ether 2.4 820 29 0.013Eugenol 1.2 30 400 0.18Farnesol 1.2 20 600 0.27Linalool 1.4 6 2300 1(-)-Rose oxide 0.46 0.5 9200 4.1(-)-Carvone 0.41 50 82 0.036Rose furan 0.16 200 8 0.003beta-Damascenone 0.14 0.009 156000 70beta-Ionone 0.03 0.007 42860 19.2

Odor Unit = ConcentrationOdor Threshold

O

CH3

CH3CH3

CH3 CH3

CH3

CH3

O

CH3

CH3 CH3

OH

CH3 CH3

CH3

O

CH3

Physical Protection

Increase/decrease Plant architecture

Leaf shapes Colours

Special features (i.e., spikes, etc)

Secondary Metabolites

Repellents Attractants

Ingestion inhibitors Taste

Camouflage odours

Plant

Signals

Direct & indirect

Dual action

Increase/decr

ease production

Vary

production & dissipation

Direct & Indirect Responses

Herbivory (Predators)

Sign

al

Resp

onse

s

Insect Elicitors

Insects

Avoid “dangerous” metabolites

Produce counter metabolites (i.e.,

digestion enzymes)

Time/ spatial avoidance

Plant Repair

Heat Drought

UV Stress Flood & water

logging Lack of nutrients

Plant damage

Environment Changes

Oxygen

Carbon Dioxide

Water

Nitrates & Mineral Salts

Glycose

Polysaccharides & Disaccharides

Glycolysis

6-Deoxyxylulose

Isoprenoids (terpenoids)

Erythrose 4-phosphate

Pentose Phosphate Pathway

Phosphoenol pyruate

Pyruvate Hydroxy-

Benzoic Acid

Shikimate Pathway

Alkaloids

Phenylpropanoids

Complex Alkaloids

KREBS CYCLE

Acetyl-CoA

Malonyl-CoA

Mevalonic Acid

Complex Isoprenoids (terpenoids) Polyketides

Flavonoids

Complex Flavonoids

Primary Metabolites (in blue) Secondary Metabolites (in red)


Amino Acids & Nucleotides

Photosynthesis

Triglycerides

Shikimate

Chorismate

Prephenate

p-hydroxybenzoate

p-aminobenzoate

Phenylalanine

Cinnamic Acids

Tyrosine

Alkaloides


Anthranilate

Tryptophan

Phenyl-C3 Compounds

Phenyl-C1 Compounds

Flavonoids

The Shikimic Acid Pathway

Acetyl CoA

3-methylglutaryl CoA (HMG-CoA)

Mevalonate

Isoprenyldiphosphate (IPP) and Dimethyllallyldiphosphate (DMAPP)

Geranyl pyrophosphate (GPP)

Geranyl geranyl pyrophosphate

(GGPP)

Other Terpenes (Triterpenes & Tetraterpenes)

C30+ (6 Isoprene Units +)

Monoterpenes

Glyceraldehyde phosphate

Pyruvate

Mevalonate Pathway

Deoxyxylulose Phosphate Pathway

Farnesylpyrophosphate (FPP)

Sesquiterpenes

C5 (1 Isoprene Unit)

C10 (2 Isoprene Units)


Diterpenes & Carotenoids


Ionones (from degraded carotenoids)

Esters

Transacylase (coenzyme)

Dihydrofolate Reductase (coenzyme)

Dihydrofolate Reductase (coenzyme)

Aldehydes & Ketones

Alcohols Acids (carboxlic)

& Acyl CoA

Sulfate

ATP Sulphurlyase

Adenylation

Adenosin 5’-phosphosulphate

(APS)

Phosphorised

APS reductase

Sulfite

Sulfide O-acetylserine (thiol)lyase

Ferrodoxin-dependent sulphite

reductase

Cysteine

O-acetylserine

3’-phosphate 5-phosphosulphate

(PAPS)

APS Kinase

PAPS Reductase

Primary Sulphur Reduction Pathway

Minor Sulphur Reduction Pathway

0

5

10

15

20

25

30

35

40

Time of the Day

Perc

enta

ge o

f Tot

al V

olat

iles

1800 2400 0600 1200 1800 2400 0600 1200 1800 2400

Figure 9.5. Chemotype Variances within the Genotype Tanacetum vulgare L.

Tansy Tanacetum vulgare L

(Asteraceae)

Genotype Chemotypes

Camphor Type

1,8-Cineole Type

Artemisia ketone Type

Thujone Type

Myrtenol Type

Chemotype Variances

Other Chemotype

Compound C1 C2 C3 1,8-Cineole 16.0 27.2 14.5 Borneol 1.8 0.1 0.8 Camphor 16.2 9.8 0.5 Terpenin-4-ol 2.4 13.8 1.9 Myrtenol 24.9 10.6 15.8 E-Nerolidol Tr. 1.7 0.6 a-pinene 5.5 4.9 0.5 ß-pinene 2.2 2.3 Tr. Spathulenol 1.3 1.7 1.8 a-thujene Tr. 0.1 Tr. a-Terpinene 0.5 3.1 0.5 Ў-Terpinene 0.9 5.6 0.3 o-Cymene 0.7 3.4 0.3 Sabinene 1.7 1.7 Tr. Camphene 1.5 0.8 1.1

Sample Linalool Methyl chavical

Olfactory Profile

India 14.2% 77.5% A grassy herbaceous and mildly spicy predominating note, with an herbaceous subsidiary note; back notes slightly fruity.

French 55.3% 10.9% A smooth fresh and diffusive herbaceous note with harmonized cool anisic and slightly balsamic subsidiary notes and warm woody back notes.

Australian 34.3% 34.7% A clean vegetable type note with a cool herbaceous menthol-like subsidiary note; a green and grassy back note.

Seychelles 27.7% 40.2% A sharp diffusive clean grassy herbaceous note, with a fruity anisic subsidiary note and a very slightly camphoraceous back note.

(Australian grown)

3.4% 75.7% A sharp, if not somewhat dry, anisic note; the subsidiary notes were herbaceous with a slight sweet camphoraceous floral back note.

Different Major Chemical and Olfactory Profiles of Five Basil Oils

Influencing Factors Compound

characteristics (volatility/mol. Weight) Surrounding Material

Distillation

Historical

Solvent Extraction

Cold Pressing

Highly volatile terpenes

Compounds mixed with waxes, also lactones, esters etc. Low/medium volatility/stable

Influencing Factors Price vis. Market

Volume Plant cell structures

Field size Topography

Soil/field characteristics Part of plant (i.e.,

rhizome/leaf) Coppice

Automated harvest and distillation

system

Harvest and later load system

Manual harvesting

due to scale,

material, investment

Extensive large scale

farming High --- Low

Value

Specialties Distillation

Hydro

Water-Steam

Steam

Material Characteristics

Material Characteristics

Solubility in water

CO2 ‘Finer’

composition spectrum

Fractional Distillation

Individual aroma

compounds

Carbon dioxide Glycolysis Glucose 2 Acetyl CoA Fatty acid Acetoacetyl-CoA thiolase Acetocetyl CoA HMGS-CoA syntesis 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) HMGL-CoA lyase HMGR-CoA reductase Mevalonate Mevalonate pyrophosphate IPP isomerase Isopentenyl pyrophosphate (IPP) DMAPP (C5) Monoterpene synthases and cyclase Geranyl pyrophosphate (C10) prenyltransferase Farnesyl pyrophosphate (FPP) (C15) Sesquiterpene Synthesis & Cyclase Diterpene synthase & Cyclase Squalene syntase Geranyl geranylpyrophosphate (C20) Squalene (C30)

Cytokinins Isopentyl adenine

rubber

Monoterpeness

Sesquiterpenes Polyprenols

Farnesylated proteins

Sterols Saponins Hormones

Lipoproteins

Diterpenes Carotenoids Abscisic acid Chlorophyll Vitamin K

IPP DMPP Isomerization Geranyl-PP (+)-3S-Linayl-PP Monoterpene cyclase (-)-Boynyl-PP (-)-Camphene (+)-Isobornyl acetate (-)-Camphor Dehydrogenase (-)-Bornyl (-)-Camphor Dehydrogenase (+)-cis-sabinol (+)-Sabinone (+)-3-Thujone (a-thujone) (-)-3-Isothujone (ß-thujone)

Harvest Timing Critical for Some Crops (Mentha piperata)

beta-Pinene

Myrcene

Linalool

Geraniol Citronellol Citronellal

MentholLinalyl Acetate

1959 – Bain & Webb – Turpentine into Fragrance & Flavor

CH3CH3

C H2

C H 2

CH 3 C H3

C H2

CH 3

CH3 C H3

C H2

O H

CH 3

CH 3 CH 3

O H

C H3

CH3 C H3

O H

C H 3

CH 3 CH3

O

C H 3

CH 3 CH 3

O H

CH3

CH 3 CH 3

CH 2

O

OCH 3

1959 – Roche Process for Linalool & Citral via Acetylene

O

OH

OH

CHO

Linalool

Citral

Points to Remember

• The highest probability is not necessarily the correct one

• Check back to see if that chemical could physically be present

• You will have to add new materials to your database (Both NIST & Wiley databases)

• Is it an artifact? • Confirm your analysis with other supportive data

Using CGMS in Quality Assurance

of Flavours & fragrances

QC lab. in the days before the concept of ‘Health & Safety at Work’ – note lighted cigarettes dangling from lips of staff!

Gas Chromatography – Mass Spectrometry

GC-MS analysis Detective work

Looking at metabolite paths Olfactory analysis of living plants

Third Nobel Prize in this field

Helps elucidate many of the C10H16 group terpene structures present in essential oils utilizing common reagents such as hydrogen chloride and hydrogen bromide. In 1909 he published the results of his extensive studies in the book Terpene und Campher, a volume of 600 pages dedicated to his pupils.

CH3

CH3CH3

CH3

CH3CH2

CH2

CH3

CH3

CH3CH3

CH3O

beta-Pinenealpha-Pinene

CampheneCamphor

No. 3: Nobel Prize in Chemistry 1910 – Otto Wallach

In the perfumery and sesquiterpene domain - the total syntheses of nerolidol and farnesol.

From Jasmine - established the structure of jasmone. Elucidated the structures of the naturally occurring

musk perfumes, civetone and muscone thus replacing scents prized since antiquity – but only available from endangered species.

OO CH3

Civetone MusconeCivet CatViverra civetta

Musk DeerMoschus moschiferus L.

Nobel Prize in Chemistry 1939 - Leopold Ruzicka

Fourth Nobel Prize in this field

Have fun with your new toy

Thank You

GCMS presentation - natural products

Technology

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