GCMS presentation - natural products


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Using CGMS as a forensic tool in discovering natural materials and creating fragrances

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GCMS presentation - natural products

  1. 1. The Metabolite Map for Identifying Probabilities and Olfactory Analysis As A Supplement to GCMS Analysis <br />Murray Hunter<br />
  2. 2. 1. Olfactory Evaluation<br />2. Using CGMS in Forensic Fragrance Construction<br />3. Using CGMS in Identifying Essential oil constituents<br />4. Using CGMS in Quality Assurance of Flavours & fragrances<br />
  3. 3. What are essential oils?<br />
  4. 4. Essential Oil<br />A volatile oil obtained from a <br />wide variety of plant, scrub, <br />and tree species and from <br />various parts of the plant <br />anatomy, such as the roots, <br />rhizomes, wood bark, leaves, <br />stems, fruit, flowers and seeds. <br />Usually extracted by hydro or <br />steam distillation, expression <br />or effleurage - Hunter 1996<br />
  5. 5. Concretes and Absolutes<br />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.<br />
  6. 6. The Natural Aromatic Product Family<br />
  7. 7. Olfactory Evaluation<br />
  8. 8. ?<br />The Olfactory System & Odour Classification<br />
  9. 9. The Human Olfactory System<br />Odour molecules pass through the olfactory organ<br />
  10. 10. Olfactory epithelium<br />Olfactory mucosa<br />Mucus!<br />High in nasal cavity<br />Site of transduction<br />Contains olfactory receptor neurons (ORN)<br />
  11. 11. 11<br />Nose hair: Olfactory cilia<br />ORN have cilia<br />Cilia contain olfactory receptor proteins<br />Similar to visual pigment<br />Transduction<br />Odorants bind to ORs<br />Change shape of protein<br />Ion flow across OR<br />Electricity<br />
  12. 12. Transmission<br />Olfactory Receptors<br />Olfactory Cells line the Olfactory Epithelium which is responsible for olfaction<br /> Each cell has cilia where receptor sites are located<br /> Replaced every month<br /> Axons of the olfactory receptors carry information to the olfactory bulb<br /> Olfactory bulb sends axons to several specific parts of the cortex with precise connections <br /> From the cortex, information is sent to other areas that control feeding & reproduction<br />Olfactory Disorder<br />Anosmia: the general lack of olfaction<br /> Specific Anosmia: the inability to smell a specific chemical<br />
  13. 13. 13<br />How many receptor types are there?<br />1000 different kinds of olfactory receptors (OR)<br />10 million OR neurons<br />10,000 of each type of OR<br />Each OR neuron has only one type of receptor<br />1000 neuronal chemical detectors<br />Potential to differentiate between 5,000-10,000 different odours<br />
  14. 14. Olfactory Bulbs<br />An outcropping of the brain<br />Its like a snail in your brain!<br />Electrical responses in cilia passed through olfactory nerve to OB<br />
  15. 15. The hedonic primacy of olfaction<br />Sensory and emotional experience<br />Not the same for vision/audition<br />Seeing and feeling more distinct<br />More intertwined in the chemical senses<br />Why?<br />Orbitofrontal cortex<br />Plays multiple roles<br />Critical for emotional<br /> experience<br />Secondary sensory <br />cortex for olfaction<br />Also centre of creativity and imagination<br />
  16. 16. The olfactory interpretation process from input to response<br />
  17. 17. Nobel Prize<br /><ul><li> 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.</li></li></ul><li>The Structure of a Fragrance<br />Top Notes<br />Bergamot oil, Rosewood oil (linalool), Linalyl acetate, Neroli oil, Ciste Oil<br />“First Impression” in Perfume.<br /> High Volatile Citrus, Fresh, Green <br /> notes…..<br />Rose Oil 5%, Jasmin absolute 4%, Ylang Ylang Oil, Aldehyde C11, C12, Methyl ionone 8%, hydroxycitronellal 10%, Cinamic Alcohol, Styrax. Phenol ethyl alcohol, Phenylacetaldehyde<br />Middle Notes<br />“ Heart of fragrance”.<br /> Intermediate Volale<br />Floral, Aldehydic notes…..<br />Vetiveryl acetate, Sandalwood, Isoeugenol, Vanillin 1.5%, Coumarin 15%, Nitromusks 10% <br />Base Notes<br />Basic of fragrance”.<br />(Bottom) Low Volatile<br />Woody, Powdery, Musky …..<br />
  18. 18. Classification of Odours<br />Curtis & Williams (1994)<br />
  19. 19. Floral Family Descriptions<br />
  20. 20. Woody Family Descriptions<br />Animalic Family Descriptions<br />
  21. 21. Balsamic Family Descriptions<br />Herbaceous Family Descriptions<br />Agrestic Family Descriptions<br />
  22. 22. Green Family Descriptions<br />Minty Family Descriptions<br />Coniferous Family Description<br />
  23. 23. Marine Family Descriptions<br />Aldehydic Family Descriptions<br />Medicated Family Descriptions<br />
  24. 24. Fruity Family Descriptions<br />Sub-Class (Citrus)<br />
  25. 25. Spicy Family Description<br />Miscellaneous Notes<br />
  26. 26. Odour Characteristics<br />Additional Characteristics<br />
  27. 27. Fragrance Description Workshop<br />
  28. 28.
  29. 29.
  30. 30. Performance Test<br />GC-MS<br />A Fragrance Development Lab<br />A trained human more accurate than a GC-MS<br />
  31. 31. Sample of a Flavour Chart<br />
  32. 32.
  33. 33. Using CGMS in Forensic Fragrance Construction<br />From This<br />To This<br />
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  35. 35.
  36. 36. Apples aren't the same<br />Sweet, sour, green, red, sharp, fresh, wholesome, etc.<br />This means different aromatic chemicals play a role in fragrance<br />
  37. 37.
  38. 38. Discovered in 1937, the First Publication on <br />Raspberry Ketone appears in 1961<br />
  39. 39. 1964 – Introduction of Furaneol®<br />1965 – First publications on isolation of Furaneol®<br />from strawberry and pineapple<br />
  40. 40. 1965-66 – α- & β-Sinensal isolated from orange oil<br />
  41. 41. 1960’s – Nootkatone<br />First isolated in 1962, it’s importance to citrus flavor was unknown until it’s isolation from grapefruit by Mcleod in 1964.<br />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.<br />(+)-Nootkatone<br />strong grapefruit odor, <br />bitter in taste <br />(-)-Nootkatone<br />weak woody (vetiver note);<br /> no grapefruit character;<br /> virtually no taste<br />Threshold: 60,000 ppb 800 ppb<br />
  42. 42. 1970’s-80’s – Other Furanones<br />Sotolon (caramel furanone) & Maple furanone identified in cane sugar – powerful caramel maple notes<br />Maple Furanone (Abhexone)<br />Sotolon<br />Threshold: 0.003 ppb 0.00005 ppb<br />Sotolon – key flavorant of fenugreek, sake, sherry wine<br />
  43. 43. 1968 – Pfizer patents Ethyl maltol<br />Ethyl maltol (Veltol Plus®) was touted to be about 6X<br />stronger than maltol and an important substitute for <br />Coumarin. Well, at least it is stronger than maltol.<br />
  44. 44. 1975 - Introduction of α-Damascone and β-Damascone<br />1982 – Introduction of Damascenone<br />1970 – Discovery Damascenone & β-Damascone<br />
  45. 45. 1974 – Thiomenthones identified in Buchu oil<br />Key component for black currant and <br />the “fuzzy” peach skin note<br />
  46. 46. 1980 – Patent on Oxane®<br />1977 - Winter identifies 2-methyl-4-propyl-1,3-oxathiane as a character impact compound in Passionfruit<br />1984 – Pickenhagen determines that (-)- cis- 2-methyl-4-propyl-1,3-oxathiane is the important diastereomer<br />(-)-(2R,4S)-2-methyl-4-propyl-1,3-oxathiane<br /><ul><li> Odor: powerful sulfury, tropical fruit
  47. 47. Threshold: 2 ppb</li></ul>(+)-(2S,4R)-2-methyl-4-propyl-1,3-oxathiane<br /><ul><li> Odor: flat, estery, camphoraceous, </li></ul> floral, less sulfury<br /><ul><li> Threshold:4 ppb</li></li></ul><li>1982 – 1-p-Menthene-8-thiol<br />Demole identifies 1-p-Menthene-8-thiol as a character impact <br />compound in Grapefruit Juice <br />Very powerful with an odor threshold of 0.0001 ppb, it also<br />requires stabilization as it tends to rapidly cyclize to the thio<br />analog of dihydropinol.<br />
  48. 48. Coumarinic p-Menthane Lactones<br />Dihydromintlactone (1995)<br />Wine lactone (1996)<br />Mintlactone (1983)<br />GRAS No. -- 3764 4032<br />Odor: Vinous, lactonic Minty, coumarinic, Coumarinic, hay, <br /> (coconut) tenacious lactonic, tonka<br />Flavor: Lactonic, sweet, herbal Coumarinic, fatty, Coumarinic, herbal lactonic, coconut<br />Threshold*: 0.0000016ppb 0.00012 ppb 0.000039 ppb<br />*In air<br />Ref: Frerot, et. al., FlavourFragr. J., 2002; 17: 218–226;<br />Gaudin, Tetrahedron, 2000, 56: 4769-4776<br />
  49. 49. 1962 – First publications appear on Methyl Jasmonate & Methyl Dihydrojasmonate<br /> <br />1R,2R-(-)-Z-methyl jasmonate 1S,2S-(+)-Z-methyl jasmonate<br /> <br />1R,2S-(+)-Z-methyl epijasmonate 1S,2R-(-)-Z-methyl epijasmonate<br />Methyl Jasmonate – odorant, insect pheromone, plant growth regulator<br />
  50. 50. Advances in Flavoring Materials<br />Chirality & Odor - Methyl Jasmonate & Methyl Dihydrojasmonate<br />ConfigurationOdor DescriptionOdor Threshold (in PPB)<br />Methyl jasmonates<br />1R,2R-(-)- Weak odor >70*<br />1S,2S-(+)- Odorless Odorless<br />1S,2R-(-)-epi- Odorless Odorless<br />1R,2S-(+)-epi- Strong odor; floral, true jasmin-like 3*<br />Ref: T.E. Acree et. al., J.Agric.Food.Chem. 1985<br />Methyl dihydrojasmonates<br />1R,2R-(-)- Floral, sweet, jasminelike 240**<br />1S,2S-(+)- Floral, fatty, cis-jasmone, hay character, 15,360**<br /> tea note, slightly lemon peellike (weak)<br />1S,2R-(-)-epi- Herbal, fatty, tea-like, tobacco, 12,500**<br /> β-damascone, cis-jasmone<br />1R,2S-(+)-epi- Intensely floral, jasmine-like, bright, cis- 15**<br /> jasmone, slightly fatty, woody, <br /> β-ionone-like, extremely long lasting<br />Ref: Werkhoff, P., et. al., Food Reviews International, 2002<br />* detection threshold<br />**recognition threshold<br />
  51. 51.                             <br />Advances in Flavoring Materials<br />2001 – Takasago’s Professor RyojiNoyori wins<br /> the 2001 Nobel Prize for Chemistry<br />
  52. 52.
  53. 53.
  54. 54. Individual Aromatic Chemicals<br />Probable Constituents of an essential oil<br />
  55. 55. GC-MS Analysis of a Meat Flavor<br /><ul><li>Abundance</li></ul>Peak Identified as FurfurylMercaptan<br />Powerful Coffee Aroma<br /> 1.6e+07<br /> 1.4e+07<br />Peaks are 2-Methyl-2,3-dihydrofuran-3-thiol isomers<br />Powerful meat-like aroma<br /> 1.2e+07<br /> 1e+07<br />8000000<br />6000000<br />4000000<br />2000000<br />0<br />12.00<br />14.00<br />16.00<br />18.00<br />20.00<br />22.00<br />24.00<br />26.00<br />28.00<br />30.00<br />32.00<br />34.00<br />36.00<br />Time--><br />
  56. 56. Basic Fragrance Types<br /><ul><li>Oriental
  57. 57. Chypre
  58. 58. Fougere</li></ul>Citrus<br />Floral<br />Aldehydic<br />Spicy<br />
  59. 59. <ul><li>Citrus notes </li></ul>Citrus + Floral<br /><ul><li>Representative Ingredients</li></ul>Natural : Lemon Oil, Bergamote, Lime….<br />Chemical : Citral, Dihydo Myrcenol….<br /> Eau Savage<br />Bergamot<br />Lemon<br />Orange <br /><ul><li>Representative Fine Fragrance</li></ul>O de Lancome (1975. Lancome)<br />Bulgari Eau Parfume (1992. Bulgari)<br />CK one (1994. C.Klein) <br />Eau Savage (1966.C.Dior)<br />Rose <br />Jasmine<br />Lily of Valley<br />Oakmoss<br />Ambergris<br />Civet<br />
  60. 60. <ul><li>Citrus notes </li></ul>CK One<br />Bergamot<br />Lemon<br />Mandarin<br />Dihydro Myrcenol <br />Citrus<br />Green<br />Floral<br />Jasmin<br />Muguet<br />Amber<br />Musk<br />Sandalwood<br />Amber<br />Woody Musk<br />
  61. 61. <ul><li>Floral notes </li></ul>Floral + Floral<br />JOY<br /><ul><li>Representative Ingredients</li></ul>Natural : Jamine Abs, Ylang Ylang, Rose Abs.<br /> Tuberose ….<br />Chemical : Hedion, Benzyl Acetate….<br />Green <br />Green <br /><ul><li>Representative Fine Fragrance</li></ul>Joy (1935. Jean Patou)<br />Diorssimo (1956.C.Dior)<br />Anais Anais (1979. Cacharel)<br />Paris (1983. YSL)<br />Beautiful (1985. E. Lauder)<br />Jasmine<br />Rose <br />Ylang Ylang<br />Violet<br />Muguet<br />Floral <br />Musk<br />Sandalwood<br />Powdery<br />Musk<br />Woody<br />Powdery<br />
  62. 62. Chanel No. 5<br />Aldehyd 10<br />Muguet<br />Ylang-Ylang <br />Floral<br />Jasmine<br />Rose<br />Carnation<br />Aldehyde C-11<br />Aldehyde C-12<br />Aldehyddic<br />Woody<br />Powder<br />Vanilla<br />Sandalwood<br />Musk<br /><ul><li>Aldehydic notes </li></ul>Floral + Aldehydic<br /><ul><li>Representative Ingredients</li></ul>Natural : Not available <br />Chemical : Aldehyde C-11, Aldehyde C-12..….<br /><ul><li>Representative Fine Fragrance</li></ul>Chanel No. 5 (1921. Chanel)<br />Calandre (1969. 1969. Rabanne)<br />First (1976. Van Cleef & Arpels)<br />
  63. 63. L'Air du temps<br />Bergamot<br />Ylang Ylang <br />Floral<br />Carnation<br />Jasmine<br />Rose<br />Iris<br />Spicy Floral<br />Sandalwood<br />Cedarwood<br />Musk<br />Woody<br />Musk<br /><ul><li>Spicy notes </li></ul>Floral + Spicy<br /><ul><li>Representative Ingredients</li></ul>Natural : Clove Buds oil, Pepper oil, Cinnamon oil.<br /> Carnation……. <br />Chemical : Eugenol, Cinnamic Aldehyde ..….<br /><ul><li>Representative Fine Fragrance</li></ul>L’air du Temps (1948. Nina Ricci)<br />Fidji (1966. Guy Laroche)<br />Egoist (1990. Chanel)<br />
  64. 64. <ul><li>Representative Ingredients</li></ul>Natural : Oak moss Abs. Vertiver, Patchouli oil<br /> Galbanum Cederwood, Sandalwood<br />Chemical : Veramoss, Iso E Super..….<br />Citrus <br />Green <br />Aldehyde<br /><ul><li>Representative Fine Fragrance</li></ul>Mitsouko (1919. Guerlain)<br />Miss Dior (1947. Christian Dior)<br />Coco (1984. Chanel)<br />Ysatis (1984. Givenchy)<br />Floral (Jasmine, <br />Rose, Ylang…)<br />Woody<br />Mossy<br />Patchouli<br />What is Chypre….?<br /><ul><li>Chypre notes </li></li></ul><li><ul><li>Chypre notes - Fine Fragrance </li></ul>Bergamot<br />Mandarin<br />Galbanum<br />AldehydC-11<br />Citrus<br />Green<br />Aldehyde<br />COCO<br />Jasmine<br />Rose <br />Ylang-Ylang<br />Muguet<br />Oriental<br />Floral<br />Amber<br />Woody<br />Mossy<br />Amber<br />Mossy<br />Patchouli<br />Mossy<br />Woody<br />
  65. 65. Citrus <br />Mandarin<br />Lemon<br />Spicy(cinamon..)<br />Floral<br />Vanilla Abs.<br />Sweet<br />Balsam<br />What is Oriental….?<br /><ul><li>Oriental notes
  66. 66. Representative Ingredients</li></ul>Natural : Vanilla, Tolu Balsam, Mandarin, Cinnamon <br />Chemical : Vanillin, Galaxolide..….<br /><ul><li>Representative Fine Fragrance</li></ul>Shalimar (1925. Guelain)<br />Obsession (1984. Calvin Klein)<br />Opium (1977. YSL)<br />Samsara (1989. Guelain)<br />Jean Paul Gaultier (1993. J.P.Gaultier)<br />
  67. 67. <ul><li>Fougere notes - Fine Fragrance</li></ul>Bergamot<br />Lavendin<br />Clove<br />Spearmint<br />Herbacious<br />Drakkar Noir<br />Muguet<br />Geranium<br />Carnation<br />Floral<br />Woody<br />Powdery<br />Powdery<br />Mossy<br />Cederwood<br />
  68. 68.
  69. 69. Using CGMS in Identifying <br />Essential oil constituents<br />
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  71. 71.
  72. 72.
  73. 73.
  74. 74. Ylang Ylang (Cananga odorata)<br />benzyl acetate (ca. 25 %), p-cresyl methyl ether (ca. 20 %), <br />methyl benzoate (ca. 5 %), methyl salicylate, cinnamyl acetate, <br />(-)-linalool (ca. 15 %), geranyl acetate (ca. 10 %), farnesyl acetate (ca. 3 %), <br />as well as a number of other sesquiterpenes and their oxygenated <br />derivatives, e.g. muurolol T (ca. 2 %)<br />Extraction Method: Steam distillation of the flowers<br />Origin: Indonesia, Madagascar<br />
  75. 75. Patchouli Oil (Pogostemon cablin)<br />There are no synthetic <br />equivalents of the <br />patchouli scent.<br />Main Constituents: (-)-patchoulol <br />(30-40 %). However, it is maintained <br />that norpatchoulenol, present in only<br /> 0.3-0.4 %, is playing a principal part<br /> in the overall odour picture.<br />(-)-patchoulol andnorpatchoulenol<br />Obtained by steam <br />distillation under <br />pressure or CO2-<br />extraction of the <br />dried leaves<br />Patchouli is mostly <br />grown in Indonesia<br />
  76. 76. Pandanus Oil (Pandanus odoratissimus)<br />phenethyl methyl etherpandanol<br />phenethyl methyl ether (<br />pandanol) (38 %), together <br />with terpinen-4-ol (19 %), <br />alpha-terpineol (8 %) and <br />phenethyl alcohol (7 %) [79]. <br />Phenethyl alcohol and its <br />derivatives are common <br />odorants in flowers<br />The flowers are hydro-distilled to yield a 'kewda attar<br />Origin: native of South East Asia and is much <br />cultivated on the Indian East Coast<br />
  77. 77. Backhousia citriodora (Lemon Myrtle)<br />High Investment to Expand Industry in Australia<br />Strong Demand as an Ingredient for tea<br />Good Crop to Grow in Most Parts of Thailand<br />Citral (geranial 51.43 %<br />and neral 42.12 % )<br />
  78. 78. Important New Compounds<br /><ul><li>Rose oil – What’s Important</li></ul>Concentration<br />Odor Unit =<br />Odor Threshold<br />
  79. 79.
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  87. 87.
  88. 88. Different Major Chemical and Olfactory Profiles of Five Basil Oils<br />
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  90. 90.
  91. 91.
  92. 92.
  93. 93. Harvest Timing Critical for Some Crops (Mentha piperata)<br />
  94. 94. 1959 – Bain & Webb – Turpentine into Fragrance & Flavor<br />beta-Pinene<br />Linalool<br />Linalyl Acetate<br />Menthol<br />Myrcene<br />Geraniol<br />Citronellol<br />Citronellal<br />
  95. 95. 1959 – Roche Process for Linalool & Citral via Acetylene<br />
  96. 96. Points to Remember<br />The highest probability is not necessarily the correct one<br />Check back to see if that chemical could physically be present<br />You will have to add new materials to your database (Both NIST & Wiley databases)<br />Is it an artifact? <br />Confirm your analysis with other supportive data<br />
  97. 97.
  98. 98. Using CGMS in Quality Assurance of Flavours & fragrances<br />
  99. 99. QC lab. in the days before the concept of ‘Health & Safety at Work’ – note lighted cigarettes dangling from lips of staff!<br />
  100. 100.
  101. 101. Gas Chromatography – Mass Spectrometry<br />Detective work<br />GC-MS analysis<br />Looking at metabolite paths<br />Olfactory analysis of living plants<br />
  102. 102. Third Nobel Prize in this field<br /><ul><li>No. 3: Nobel Prize in Chemistry 1910 – Otto Wallach
  103. 103. 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.</li></ul>beta-Pinene<br />alpha-Pinene<br />Camphene<br />Camphor<br />
  104. 104. Fourth Nobel Prize in this field<br /><ul><li>Nobel Prize in Chemistry 1939 - Leopold Ruzicka
  105. 105. In the perfumery and sesquiterpene domain - the total </li></ul> syntheses of nerolidol and farnesol. <br /><ul><li>From Jasmine - established the structure of jasmone.
  106. 106. Elucidated the structures of the naturally occurring </li></ul> musk perfumes, civetone and muscone thus replacing <br /> scents prized since antiquity – but only available from <br /> endangered species.<br />Civetone<br />Muscone<br />Musk Deer<br />Moschusmoschiferus L.<br />Civet Cat<br />Viverracivetta<br />
  107. 107. Have fun with your new toy<br />Thank You<br />