Practical Analysis of Flavor and Fragrance Materials Practical Analysis of Flavor and Fragrance Materials Edited by Kevin Goodner and Russell Rouseff A John Wiley & Sons, Ltd., Publication This edition first published 2011 © 2011 Blackwell Publishing Ltd Registered office John Wiley & Sons Ltd, The Atrium, Southern Gate, Chichester, West Sussex, PO19 8SQ, United Kingdom For details of our global editorial offices, for customer services and for information about how to apply for permission to reuse the copyright material in this book please see our website at www.wiley.com The right of the author to be identified as the author of this work has been asserted in accordance with the Copyright, Designs and Patents Act 1988 All rights reserved No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, except as permitted by the UK Copyright, Designs and Patents Act 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978-1-119-97522-9 Typeset in 10.5/13pt Sabon by Laserwords Private Limited, Chennai, India Contents Preface xiii About the Editors xvii List of Contributors xix Overview of Flavor and Fragrance Materials David Rowe 1.1 1.2 1.3 Flavor Aroma Chemicals 1.1.1 Nature Identical 1.1.1.1 Alcohols 1.1.1.2 Acids 1.1.1.3 Esters 1.1.1.4 Lactones 1.1.1.5 Aldehydes 1.1.1.6 Ketones 1.1.2 Heterocycles 1.1.2.1 Oxygen-containing 1.1.2.2 Nitrogen-containing 1.1.2.3 Sulfur-containing 1.1.3 Sulfur Compounds 1.1.3.1 Mercaptans 1.1.3.2 Sulfides Flavor Synthetics Natural Aroma Chemicals 1.3.1 Isolates 1.3.2 Biotechnology 1.3.3 ‘Soft Chemistry’ 1 5 6 8 9 10 11 12 12 13 vi CONTENTS 1.4 1.5 Fragrance Aroma Chemicals 1.4.1 Musks 1.4.2 Amber 1.4.3 Florals 1.4.4 ‘Woodies’ 1.4.5 Acetals and Nitriles Materials of Natural Origin 1.5.1 Essential Oils 1.5.1.1 Cold-pressing – Citrus Oils 1.5.1.2 Steam-distilled Oils 1.5.1.3 A Note on ‘Adulteration’ 1.5.2 Absolutes and Other Extracts Acknowledgments References Sample Preparation Russell Bazemore 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 Introduction PDMS Static Headspace Extraction 2.3.1 Advantages and Disadvantages Dynamic Headspace Extraction 2.4.1 Advantages 2.4.2 Disadvantages Solid Phase Microextraction (SPME) 2.5.1 Research 2.5.2 Practical 2.5.3 Advantages 2.5.4 Disadvantages Stir Bar Sorptive Extraction 2.6.1 Research 2.6.2 Practical 2.6.3 Advantages 2.6.4 Disadvantages PDMS Foam and Microvial 2.7.1 PDMS Foam 2.7.2 Microvial Solvent Extraction 2.8.1 MIXXOR 2.8.2 Soxhlet Extraction 14 14 15 16 17 17 18 18 18 19 20 21 21 21 23 23 24 25 25 26 27 27 27 27 29 32 32 33 33 34 35 35 36 36 36 39 39 39 CONTENTS 2.9 2.8.3 Solvent Assisted Flavor Evaporation (SAFE) Summary References Traditional Flavor and Fragrance Analysis of Raw Materials and Finished Products Russell Rouseff and Kevin Goodner 3.1 3.2 3.3 Overview Physical Attribute Evaluation 3.2.1 Color – Optical Methods 3.2.2 Turbidity 3.2.3 Water Activity 3.2.4 Moisture Content 3.2.4.1 Karl Fischer Method 3.2.4.2 Secondary Moisture Determination Methods 3.2.5 Optical Rotation 3.2.6 Specific Gravity 3.2.7 Refractive Index 3.2.8 Sugars/Soluble Solids 3.2.9 Viscosity Instrumental Analysis 3.3.1 Separation Techniques 3.3.1.1 Gas Chromatography (GC) 3.3.1.2 GC Retention Data 3.3.1.3 Standardized Retention Index Systems 3.3.1.4 GC Injection 3.3.1.5 GC Columns (Stationary Phases) 3.3.1.6 GC Detectors 3.3.2 Identification Techniques 3.3.2.1 Retention Index Approach 3.3.2.2 GC–MS 3.3.2.3 MS/MS References Gas Chromatography/Olfactometry (GC/O) Kanjana Mahattanatawee and Russell Rouseff 4.1 4.2 4.3 Introduction Odor Assessors’ Selection and Training Sensory Vocabulary vii 42 42 42 45 45 47 48 49 49 50 50 51 51 52 52 53 54 54 55 55 55 55 56 58 60 63 63 64 65 67 69 69 70 71 viii CONTENTS 4.4 4.5 4.6 4.7 4.8 4.9 GC/Olfactometers (Sniffers) Practical Considerations Types of GC/Olfactometry 4.6.1 Dilution Analysis 4.6.2 Time Intensity 4.6.3 Detection Frequency 4.6.4 Posterior Intensity Method Sample Introduction Identification of Aroma-active Peaks 4.8.1 Standardized Retention Index Values 4.8.2 Aroma Description Matching 4.8.3 MS Identifications 4.8.4 Use of Authentic Standards Conclusion References Multivariate Techniques Vanessa Kinton 5.1 5.2 5.3 5.4 5.5 5.6 Introduction Hierarchical Cluster Analysis (HCA) Principal Component Analysis (PCA) Classification Models 5.4.1 k-Nearest Neighbors (k-NN) 5.4.2 Soft Independent Modeling of Class Analogy (SIMCA) Principal Component Regression Example of Data Analysis for Classification Models 5.6.1 Tabulating Data 5.6.2 Examining Data 5.6.3 Multivariate Exploratory Analysis 5.6.4 Creation of a Classification Model with a Training Set and Validation with a Testing Set References Electronic Nose Technology and Applications Marion Bonnefille 6.1 6.2 6.3 Introduction Human Smell and Electronic Noses Techniques to Analyze Odors/Flavors 6.3.1 Sensory Panel 6.3.2 GC and GC/MS 72 73 73 73 76 79 82 83 84 84 85 85 86 86 87 91 91 97 98 99 100 100 101 102 102 103 103 106 109 111 111 112 113 113 114 CONTENTS 6.3.3 6.3.4 6.3.5 6.4 6.5 GC/Olfactometry Electronic Nose Electronic Nose Technology and Instrumentation 6.3.5.1 Architecture 6.3.5.2 Air Generator 6.3.5.3 Sampling 6.3.5.4 Detection Technologies 6.3.6 Data Treatment Tools The Main Criticisms Directed at the Electronic Nose Market and Applications 6.5.1 Application Range 6.5.2 Perfumery Compound Detection in a Fragrance 6.5.3 Cosmetic Natural Raw Materials: Characterization of Volatile Constituents of Benzoin Gum 6.5.4 Home Care Products: Identification and Quantification Using an Electronic Nose in the Perfumed Cleaner Industry 6.5.5 Pharmaceutical Products: Flavor Analysis in Liquid Oral Formulations References MS/Nose Instrumentation as a Rapid QC Analytical Tool Ray Marsili 7.1 7.2 7.3 7.4 7.5 Introduction Operating Principle Advantages of MS over Solid State Sensors Using Other Sample Preparation Modes Techniques for Improving Reliability and Long-term Stability 7.5.1 Calibration Transfer Algorithms 7.5.2 Internal Standards 7.6 Two Instruments in One 7.7 Application Examples 7.8 Classification of Coffee Samples by Geographic Origin 7.9 Classification of Whiskey Samples by Brand 7.10 Future Directions: Partnering MS/Nose with GC/MS 7.11 Conclusion References ix 114 115 115 115 117 118 121 127 134 136 136 138 139 142 146 151 155 155 157 160 160 161 161 162 163 163 164 166 168 170 170 x CONTENTS Sensory Analysis Carlos Margaria and Anne Plotto 8.1 8.2 8.3 8.4 8.5 8.6 8.7 8.8 8.9 Introduction The Purpose of Sensory Analysis Flavor Perception Sensory Analysis Techniques 8.4.1 Overall Difference Tests 8.4.1.1 Triangle Test 8.4.1.2 Duo–Trio Test 8.4.1.3 Simple Difference Test 8.4.2 Single Attribute Difference Tests 8.4.2.1 Difference from Control 8.4.2.2 Paired Comparison Test 8.4.2.3 Ranking Tests 8.4.3 Descriptive Tests 8.4.4 Affective Tests Preparation and Planning 8.5.1 Experimental Design 8.5.2 Environment 8.5.3 Sample Preparation Panel Selection 8.6.1 Trained Panels 8.6.2 Consumer Panels Conducting a Panel Expression of Results Conclusions References Regulatory Issues and Flavors Analysis Robert A Kryger 9.1 9.2 Introduction Regulatory Overview 9.2.1 History 9.2.2 Safety Regulations 9.2.3 Product Labelling Regulations 9.2.4 Fair Trade/Conformity with Established Standards 9.2.5 Flavor Types 9.2.6 Governing Authorities 9.2.7 Role of Flavor Analysis in Regulatory Conformance 173 173 174 177 178 179 180 182 182 184 184 184 185 186 188 189 189 191 191 192 193 194 195 196 197 198 201 201 202 202 204 206 208 209 211 212 214 PRACTICAL ANALYSIS OF FLAVOR AND FRAGRANCE MATERIALS on flavors (88/388/EEC) specifically limits lead (Pb) content in flavoring products to 10 mg/kg In the case of a well-defined chemical substance, a suitable test method is usually easy to develop with the right equipment Sometimes a standard method is available, having been developed for other food products For more difficult measurements, industry groups sometimes collaborate to develop suitable test methods [22] However, complications can arise for a number of reasons For example, a regulated substance may not be well-defined chemically Allergens provide a good example of this problem Allergen ingredients which are not declared on the label are prohibited in food blends Accidental allergen contamination, due to process equipment or raw material contamination, is another concern However, a test for ‘peanut content’ is not really a well-defined request to an analytical chemist With many different peanut byproducts, what exactly constitutes a ‘peanut’ from a chemical perspective? Test sensitivity is another potential complication Many pesticides are approved for use on specific food products only Approved pesticide usage rates may lead to the presence of pesticide residues on food products in the part per million (ppm) to part per billion (ppb) range If a sample is tested with a method sensitive to 0.1 ppb, and the result is clean, it would seem to affirm that no unapproved pesticide was used However, if a pesticide test is much more sensitive than that, how does one interpret a positive result of 0.002 ppb for an ‘unapproved’ pesticide? The presence of such a low level could be the result of inadvertent exposure during the growing and/or production of the food product Determining if this is a regulatory concern is a difficult question 9.3.1.1 Heavy Metals such as Pb, As, Hg, and Cd Heavy metal content of food and flavoring products is a relatively straightforward example of a limited contaminant Heavy metals are limited in food products due to health concerns associated with metal accumulation The allowed limits are generally well defined either in the flavoring products themselves or in the finished food products Furthermore, the technology for heavy metal testing is well established [23] and the interpretation of test results is straightforward 9.3.1.2 Pesticides Pesticides, fungicides, miticides, and related products used to improve agricultural yields are very well defined from a regulatory aspect because REGULATORY ISSUES AND FLAVORS ANALYSIS 215 these products must satisfy a very detailed government registration and approval process prior to use in most of the world As part of this process, limits are established for the use of these chemicals on various agricultural products and tests for the residues of these chemicals are established In fact, for most pesticide-type products, regulatory/agency approved analysis methods exist because analysis methods must be submitted with the pesticide registration [24] As discussed above, the most difficult regulatory questions concern the interpretation of very low pesticide levels, especially if a particular pesticide is ‘not allowed’, as well as the applicability of regulatory pesticide limits on agricultural byproducts For example, whole fruit pesticide limits exist in the US on citrus fruits [25] How you interpret those limits with regard to fruit byproducts such as peel comminute and/or peel oils? 9.3.1.3 Environmental Toxins The presence of certain naturally occurring environmental toxins – like patulin in apple juice, aflatoxin in cereals, or spice and nut products – is a more complex example Obvious natural toxins (such as the alkaloids in certain mushrooms) were identified a long time ago by their acute symptoms after consumption New toxins, or suspected toxins, are constantly being brought to regulators’ attention by health and/or food researchers based upon long-term health concerns such as carcinogenicity These toxins often occur at very low levels and regulators can feel pressure to act prior to the development of robust analytical methods Some examples include patulin in apple products and acrylamide in fried foods [26] Patulin testing in apple products is an illustrative example Patulin is a naturallyoccurring toxin generated by molds which can grow on fruit surfaces like apples [27] Patulin is suspected of causing carcinogenic or mutagenic toxicity in humans [28] When contaminated apples are processed the toxin can contaminate the resulting juice and juice by-products Any of these products which are incorporated into a flavoring mixture are a potential source of patulin Regulatory limits for patulin are in the neighborhood of 25–50 ppb [29] The development of a robust test by HPLC for patulin was complicated because preconcentration is required and patulin can be unstable under some common conditions Concentrated efforts by industry groups and others have lead to the development of acceptable methods [22] However, prior to these developments, it was difficult to obtain consistent results from different laboratories 216 PRACTICAL ANALYSIS OF FLAVOR AND FRAGRANCE MATERIALS 9.3.1.4 Allergen Testing Allergic reactions to specific food products in some individuals have been known for a long time Regulations that require the label declaration of common allergens have been implemented to address this safety issue However, the ability to test for allergen content in food or flavor products has been difficult Our knowledge of the specific agents that cause food allergies is still limited Also, the concentration levels at which food allergens are dangerous in food is not well established Both factors likely vary among affected individuals Also complicating the regulatory picture is whether components of an allergenic material are equally dangerous Does the oil from soybeans contribute to the allergenic reaction of soybean? Research seems to indicate that most food allergies result from proteins While there is little economic reason not to declare known allergens on the label, accidental contaminations due to carry-over on the processing equipment used to manufacture the final product or any ingredient is a big concern Recently, methods that detect specific proteins from known allergens have been developed to test products and process equipment rapidly [30] Unfortunately, these tests are available for only a few allergens and not enough testing has been done to establish their complete effectiveness 9.3.2 Testing Whether a Product is ‘Natural’ or Meets a ‘Standard of Identity’ A more challenging question for the analytical chemical chemist is to determine if a ‘natural flavor’ is truly natural In this case, one is usually confronted with determining whether a complex mixture of chemicals contains only ingredients which conform to the applicable regulations concerning natural ingredients and that the ingredients were processed in an acceptable fashion A similar problem confronts the question of whether a flavor meets the ‘standard of identity’ for a particular end use For example, FTNF flavors which are suitable for use without label declaration in products like fruit juices must be made only from ingredients derived from the named fruit Determining whether a complex apple flavor is derived only from apple by-products can be a challenging task In practice, the motivation to adulterate is almost always economic and the testing can be limited to ingredients where there is a large difference in cost between natural and artificial sources For example, flavor chemicals in the ‘green note’ class, such as hexanal, trans-2-hexenal REGULATORY ISSUES AND FLAVORS ANALYSIS 217 and cis-3-hexenal, are used frequently in natural fruit flavors Prior to the development of viable biofermentation routes to synthesize ‘natural’ versions of these compounds [31], they were isolated from plant extracts such as mint oils, usually at very low concentration Collection and purification was expensive, leading to prices for these natural chemicals at thousands of dollars per kg At the same time, artificial green notes derived from petrochemicals were available at a fraction of the cost Naturalness verification for green note compounds was routine In general three main approaches are used First, the identification of compounds which are not found in the natural products is often a simple marker of an adulterated flavor For example, until recently a key sulfur note from grapefruit – 1-p-methene-8-thiol – was not available except as a synthetic chemical or in very dilute form within grapefruit oils This chemical is relatively easy to detect by GC/MS and the presence of a significant quantity of this grapefruit thiol was a marker for synthetic adulteration Another example is the use of synthetic cooling agents in mint flavors Some major flavor companies have developed very intense ‘cooling agents’ that are much stronger than natural menthol Examples include Frescolat by Symrise and TK-10 by Takasago [32] Many of these compounds are not naturally occurring and therefore are an easy marker for an artificial ingredient One complication to keep in mind is the presence of trace levels of some synthetic chemicals – such as solvents like hexane – can be introduced as a result of processing aids during the production process Processing aids are perfectly acceptable in the manufacturing of natural flavors A second main strategy is the identification of trace compounds that should or should not be present based upon the available sources of natural products In this case, detailed knowledge on the source of the natural flavoring ingredients is needed by the analyst If an important flavor ingredient is only available from a limited number of natural sources, then knowledge of the usual chemical composition of the natural source is very helpful For example, an FTNF apple flavor suitable for use in apple juice must be made from ingredients entirely derived from apple One important ester present in apple aroma, a key flavor ingredient of FTNF flavors, is ethyl-2-methyl butyrate (E2MB) In natural apple, the level of E2MB is much less than the other volatile ethyl esters If this ratio is found to be unusually high in an apple flavor, it is an indication that E2MB from a nonapple source has likely beenadded Natural vanilla extract is another example Trace levels of p-hydroxybenzoic acid and vanillic acid in the right ratios with vanillin serve as indicators of natural vanilla extract [33] 218 PRACTICAL ANALYSIS OF FLAVOR AND FRAGRANCE MATERIALS Finally, the most sophisticated test strategy is a comparison of physical properties of individual molecules which differ depending upon the natural or artificial source A simple example is in the case of flavor components which are chiral – coming in both a left- and right- handed geometric isomer Often, the naturally occurring source favors one form over the other, while synthetic versions are equally distributed in both forms (‘racemic’) This was recognized early on in the case of citrus oils which are mostly composed of the limonene in the righthanded or (+)-limonene form An old test for adulteration with less expensive (-)-limonene was to measure the optical rotation of the oil Chiral molecules rotate polarized light differently depending upon the enantiomer Measuring the rotation factor for a citrus oil provided a fast estimate of the (+)-limonene content Currently, the development of chiral columns for GC analysis and the sophistication of GC equipment allow the measurement of the chiral ratio of individual molecules Therefore, the enantiomeric ratio of the E2MB within an apple flavor can be used to determine if the source is natural or artificial This type of measurement can also distinguish between different natural sources of compounds For example, we find the enantiomeric ratio of a compound like β-pinene differs depending upon the source, even for closely related plants In the case of citrus oils, β-pinene in lemon oil has 4–7 % in the (+) isomer, while mandarin oil is around 98 % (+) isomer [34] As this type of equipment becomes more readily available, tables of typical enantiomeric ratios for important chiral compounds from various natural sources are being published Natural and synthesized compounds can also differ due to the levels of certain isotopes found in the compound For example, naturally occurring flavor molecules, often derived from plants, incorporate 14 C at levels associated with the amount of 14 C present during plant metabolism However, since the half-life of 14 C is around 5700 years, the same molecules derived from petroleum by-products have much less 14 C present due to the age of the petroleum Carbon-14 testing has been heavily utilized to distinguish natural and artificial flavor molecules Carbon-14 was one of the earliest isotopes tested because of the ease it can be counted using scintillation detectors since it is naturally radioactive Other isotopes can also be used For example, 13 C, H (tritium) or 15 N vary in some molecules depending upon the different natural sources [35] Testing of nonradioactive isotopes is more difficult, usually involving some sort of high-resolution mass spectrometry with or without chemical derivitization Coupling high-resolution mass spectrometry with gas chromatography allows online determination of isotopes for REGULATORY ISSUES AND FLAVORS ANALYSIS 219 many individual flavor components nearly simultaneously Accelerator mass spectrometry (AMS) has also been applied to this problem Many examples are available in the scientific literature However, the equipment and expertise necessary to these types of measurements are very expensive and limited to a few laboratories Few companies and other organizations can maintain this type of equipment internally Often, these measurements are outsourced to specialized laboratories Even more recently, the development of SNIF–NMR [36], a method that utilizes NMR technology to determine the isotope ratio for specific locations within a given molecule, gives even more sensitivity For example, ethanol contains two carbon molecules A mass spectrometry based measurement of the 13 C-to-12 C ratio on ethanol will average over both carbons in the molecule However, NMR can be used to measure this ratio on a specific carbon – say the one attached to the OH group This more detailed information can shed even more light on the source of important molecules However, the equipment cost is at least an order of magnitude higher, requiring very specialized laboratories to perform One example of SNIF–NMR applied to flavor ingredients is vanillin [37] Clearly, naturalness testing generally requires substantial knowledge on the part of the analyst regarding the type of contaminants and adulterants to expect Furthermore, each flavor type has different issues It is difficult for regulatory agencies to possess this detailed knowledge Practically speaking, this type of information usually resides only in specialty laboratories and within the flavor industry itself 9.3.3 Testing for Other Regulatory Compliance Requirements Confirming regulatory compliance with other flavor categories – like GMO-free or country of origin labeling– involve many of the same issues as discussed above GMO verification involves measuring trace levels of particular proteins found only in the GMO raw material For most flavor formulations, any protein content is incidental to the formulation so that testing is not likely to be very effective Country of origin testing, of theoretical interest for some labeling requirements, is practically of little interest for flavors which are generally used in small quantities in the finished product Even more difficult cases involve verification of organic or kosher status for flavors Since noncompliance with these requirements does not necessarily change the chemical makeup of the flavor, verification by analysis is difficult At best, one can look for forbidden substances in the product Practically speaking, verification is often done by outside groups auditing the production process 220 PRACTICAL ANALYSIS OF FLAVOR AND FRAGRANCE MATERIALS To conclude, flavor manufacturers today has a broad array of regulations and requirements that must be met in the production and sale of their products These requirements influence the ingredients, the process, the distribution, and the final use of flavor mixtures It is of interest to the manufacturers, competing manufacturers, regulatory agencies, and the consumer that these regulations and requirements are met In this quest, analytical chemistry as applied to flavor mixtures plays an important role While flavors are mostly composed of volatile chemicals, suitable for gas chromatographic analysis, a much broader range of analytical tools must be applied to this problem A knowledgeable analytical chemist is also indispensable Continuing research in the tools and methods of flavor analysis will continue to open opportunities in this field REFERENCES Regulation (EC) No 2232/96 of the European Parliament and of the Council of 28 October 1996 laying down a Community procedure for flavouring substances used or intended for use in or on foodstuffs OJ L 299, 23.11.1996, pp 1–4, http://ec.europa.eu/food/food/chemicalsafety/flavouring/index_en.htm (accessed 31 August 2006) Administration of Food Safety (2003) http://www.ffcr.or.jp/zaidan/FFCRHome.nsf (accessed 31 August 2006); Japanese Food Chemical Research Foundation (2006) http://www.ffcr.or.jp/zaidan/FFCRHome.nsf/pages/list-desin.add-x (accessed 27 April 2011) Substances Generally Regarded as Safe (2005) 21 CFR Part 182 with periodic updates published in the journal Food Technology Most recent update Food Technology, 59, 24 Health Canada (2006) The Food and Drugs Act – Part B, Division and Division 10, http://laws-lois.justice.gc.ca/eng/acts/F-27/ (accessed 27 Apr 2011) Regulation (EC) No 852/2004 of the European Parliament and of the Council of 29 April 2004 on the hygiene of foodstuffs CS.ES Chapter 13, Volume 34, pp 319– 337 Codex Alimentarius Commission (2003) Recommended International Code of Practice General Principles of Food Hygene CAC/RCP 1-1969, Rev Codex Alimentarius Commission (2005) General Standard for Food Additives CAC/STAN 192-1995, Rev Food and Drug Administration (2004) Federal Food, Drug, and Cosmetic Act http://www.fda.gov/opacom/laws/fdcact/fdctoc.htm (accessed 31 August 2006) Current Good Manufacturing Practice in Manufacturing, Packing, or Holding Human Food 21 CFR Part 110 Revised 2010 10 Food Standards Australia and New Zealand (FSANZ) (2002) User Guide to Flavorings and Flavor Enhancers, http://www.foodstandards.gov.au/thecode/ assistanceforindustry/userguides/index.cfm (accessed 31 August 2006) REGULATORY ISSUES AND FLAVORS ANALYSIS 221 11 Council Directive 88/388/EEC of 22 June 1988 on the approximation of the laws of the Member States relating to flavourings for use in foodstuffs and to source materials for their production OJ L 184, 15.7.1988, pp 61–66 12 Japanese Food Chemical Research Foundation (2006) List of Plant or Animal Sources of Natural Flavorings, http://www.ffcr.or.jp/zaidan/FFCRHome.nsf/pages/listnat.flavors (accessed 27 Apr 2011) 13 Codex Alimentarius Commission (1987) General requirements for natural flavourings CAC/GL 29-1987 14 Food Labelling Foods: Labelling of Spices, Flavorings, Colorings and Chemical Preservatives 21 CFR Part 101.22 Revised 2010 15 Drawback on Taxpaid Distilled Spirits Used in Manufacturing Nonbeverage Products 27 CFR Part 17 (1996) Alcohol and Tobacco Trade and Tax Bureau Industry Circulars and Rulings for Manufacturers Non-Beverage Products, http://www.ttb.gov/industrial/mnbp.shtml (accessed 31 August 2006) 16 Health Canada (2006) The Food and Drugs Act - Part B, Division 2, http://lawslois.justice.gc.ca/eng/acts/F-27 (accessed 27 Apr 2011) 17 Distilled Spirits – Standards of Identity, 27 CFR Part 5.22 Wines – Standards of Identity 27 CFR Part 4.21 Use of Ingredients Containing Alcohol in Malt Beverages; Processing of Malt Beverages 27 CFR Part 7.11 18 Council Directive 2001/112/EC of 20 December 2001 relating to fruit juices and certain similar products intended for human consumption OJ L 10, 12.1.2002, pp 58–66 19 Canned Fruit Juices 21 CFR Part 146 20 Health Canada (2006) The Food and Drugs Act - Part B, Division 10 http://lawslois.justice.gc.ca/eng/acts/F-27 (accessed 27 Apr 2011) 21 Food Dressings and Flavorings 21 CFR Part 169 22 AOAC Official Method 995.10 (1996) Patulin in apple juice J AOAC, 79, 451 23 US Pharmacopeia (2003) Food Chemical Codex, 5th edn, National Academies Press 24 US Environmental Protection Agency (2006) Residue Analytical Methods http://www.epa.gov/pesticides/science/index.htm (accessed 31 August 2006) 25 Tolerances and Exemptions from Tolerances for Pesticide Chemicals in Food 40 CFR Part 180 26 US Food and Drug Administration (2004) FDA Action Plan for Acrylamide in Food, http://www.fda.gov/Food/FoodSafety/FoodContaminantsAdulteration/ ChemicalContaminants/Acrylamide/ucm053519.htm (accessed 27 Apr 2011) 27 Harrison, M A (1989) Presence and stability of patulin in apple products: a review J Food Safety, 9, 147–153 28 IARC (1986) Patulin IARC Monog Eval Carcinog Risk Chem Humans, 40, 8398; and WHO (1990) Patulin WHO Food Addit Ser., 26, 143–165 29 US Food and Drug Administration (2000) Patulin in apple juice, apple juice concentrates and apple juice products Fed Register, 65, 37791– 37792 30 Health Canada (2006) Allergen detection methods – the compendium of food allergen methodologies, http://www.hc-sc.gc.ca/fn-an/res-rech/analy-meth/allergen/ index_eng.php (accessed 27 Apr 2011) 31 Fabre, C and Goma, G (1999) A review of the production of green notes Perfumer and Flavorist, 24, 32 Erman, M (2004) Progress in physiological cooling agents Perfumer and Flavorist, 29, 34 222 PRACTICAL ANALYSIS OF FLAVOR AND FRAGRANCE MATERIALS 33 IOFI (2000) Information Letter 1271 – Authenticity of Natural Vanilla Products 34 Dugo, G et al (2001) Enantiomeric distribution of volatile components of citrus oils by MDGC Perfumer and Flavorist, 26, 20 35 Schmidt, C.O et al (2001) Stable isotope analysis of flavor compounds Perfumer and Flavorist, 26, 36 Site-Specific Natural Isotope Fractionation – Nuclear Magnetic Resonance Trademark of Eurofins Laboratories, Nantes, France Eurfins SNIF-NMR – A Unique Method to Prove Authentic Origin, http://www.eurofins.com/food-testing/foodanalyses/snif-nmr/en (accessed 31 August 2006) 37 Tenailleau, E J., Lancelin, P., Robins, R J., and Akoka, S (2004) Authentication of the origin of vanillin using quantitative natural abundance 13C NMR J Ag Food Chem., 52, 7782– 7787 Index absolutes, 21 acceptance testing, 188–9 acetals, 17–18 Acetobacter spp., 13 acids, 3–4 adulteration, 20–1, 92, 216–17 see also contaminants AEDA, 74 affective tests, 188–9 Agilent ChemSensor see ChemSensor Agilent Technologies, 155–6 alcoholic beverages, 209 alcohols, 2–3 from fermentation, 12–13 aldehydes, allergens, 214, 216 allergies, 194–5 ambergris, 15–16 ambers, 15–16 anosmia, 177 apple products, 215 aroma chemicals natural, 11–14 regulation, 203 aroma extract dilution analysis (AEDA), 74 autoscaling, 95 banned substances, 213–16 benzoin gums, 140–1, 142, 143 benzyl alcohol, beverages, 137 bread aroma, butyric acid, carbon-14 testing, 218 Carboxen, 29 cassia oil, celestolide, 14, 15 charm analysis, 74–5 ChemSensor, 155–7 applications, coffee sample classification, 164–6 calibration, 159, 161–2 modes of operation, 163 operation, 157–9 sample preparation, 160–1 cherry flavor, chirality, 218 cinnamon oil, citral, 5, 12, 203 citrus oils, 4, 18–19 classification models, 99–101 data tabulation, 102–3 Practical Analysis of Flavor and Fragrance Materials, First Edition Edited by Kevin Goodner and Russell Rouseff © 2011 Blackwell Publishing Ltd Published 2011 by Blackwell Publishing Ltd 224 coffee, 159 classification by origin, 164–6 GC analysis, 62 color evaluation, 48–9 concretes, consumer sensory panels, 194–5 contaminants, 205 see also adulteration cooling agents, 217 cosmetics, 137–8, 140–1 coumarin, databases see libraries delta-decalactone, density measurement, 52 difference from control testing, 184 difference tests overall duo-trio test, 182 simple, 182–4 triangle, 180–2 two out of five, 180–1 single attribute difference from control, 184 paired comparison, 184–5 dilution analysis, 83 dimethyl sulfide (DMS), 9–10 discriminant factorial analysis (DFA), 129–30, 140–1, 143 distilled lime oil, 20 divinyl benzene, 29 duo-trio test, 182 Durian fruit, dynamic headspace extraction, 26–7 electrochemical cell detectors, 126–7 electron impact ionization, 64–5 electronic noses, 115 advantages, 170 air generators, 117–18 applications, 136–8 coffee sample classification, 164–6 cosmetics, 140–1 home care products, 141–2 perfumery, 138–9 pharmaceuticals, 146–51 whiskey classification, 166–8 calibration, 159, 161–2 INDEX data analysis, 127–35 multivariate, 128–31 detectors, 125 electrochemical cell, 126–7 metal oxide, 121–3 MOSFET, 124–6 photoionization, 126 quartz microbalance, 124 solid-state vs MS, 156, 158–9, 160 surface acoustic wave, 124 disadvantages, 135–6 sampling, 118–19 sensory panels and, 148 system architecture, 115–17 training, 127–8 see also ChemSensor; FOX essential oils, 11–12 adulterated, 20–1 cold-pressed, 18–19 steam-distilled, 19–20 esters, ethanol, ethyl butyrate, ethyl-2-methyl butyrate, 217 EU Flavoring Directive 88/388/EEEC, 205, 213–14 eucalyptol, fair trade regulations, 208–9 fenugreek lactone, fermentation, 12–13 flavor compounds matrix interactions, 178 natural, 210 natural and artificial (N and A), 209–10 nature identical, 210 acids, 3–4 alcohols, 2–3 aldehydes, heterocycles, 6–8 ketones, lactones, 4–5 mercaptans, 8–9 sulfides, 9–10 regulation, 203–4, 206, 209–10 flavor types, 209–10 natural and synthetic, 207 225 INDEX synthetic see synthetic flavors traditional analysis, 45–6 flavor profile, 186–7 flavor testing, 177–8 florals, 16–17 fluidity, 54 Food and Drug Administration (FDA), 203–4 forbidden substances, 213–16 FOX electronic nose, 117, 140–1, 146–51 fragrances compound detection, 138–9 see also perfumery free choice profiling, 188 FTNF flavors, 217 furfurals, 6–7 trans-2-hexenal, trans-2-hexenol, hierarchical cluster analysis (HCA), 104–5 homofuranol, HSSE see headspace extraction hue index, 48 human smell, 112–13 humidity, 135–6 gas chromatography (GC), 55, 59, 114 detectors, 60–1, 62 injection, 56–8 mass spectrometry (GC/MS), 65–7, 114, 163 compared with e-nose technology, 157–8 MS/Nose and, 168–70 olfactometry, 69–70 retention index, 55–6, 63–4 sample preparation see sample preparation stationary phase, 24–5, 58–60 genetically modified organisms (GMO), 207–8, 210, 219 Georgywood, 17 geranial, Gerstel Twister, 33, 34–5 GMO free foods, 210 goodness values, 100–1, 107–8 green note flavors, jell-O, 177 The Jungle, 203 halal foods, 208 ham, 190 headspace sampling, 83, 119 compared to SPME, 32 dynamic, 26–7 PDMS foam, 36, 37 static, 25–6, 160–1 heavy metals, 214 heterocyclic compounds, 6–8 cis-3-hexenal, 2, 217 97–8, identity standards, 208–9, 216–17 Infometrix, 158–9 intensity (color), 48 intolerances, 194–5 Iso E Super, 17 ITEX, 119 k-nearest neighbor, 99–100, 106–7, 164–5, 167 karl Fischer titration, 50–1 ketones, kosher products, 208 ´ index, 56 Kovats LAB color space, 48 labeling, 206–8 lactones, 4–5, 13 libraries gas chromatographic linear retention index, 64 mass spectrometric, 65 Lily of the Valley, 16 linear retention index (LRI), 56, 63–4, 64 loss on drying, 50 maltol, mangones, 139 maple lactone, mass spectrometry, 85–6 electronic nose see ChemSensor tandem with gas chromatography, 64–5 Matrixx, 157 menthol, mercaptans, 8–9 metal oxide detectors, 121–3 226 metal oxide semiconductor field-effect transistors (MOSFET), 124–6 1-p-methene-8-thiol, 217 methyl N-methylanthranilate, methyl salicylate, 2-methylbutyrate, 4-methyloctanoic acid, microorganisms, 50 microvial extraction, 36–7, 36–9 milk, 157, 168 Minolta CR-400 colorimeter, 49 MIXXOR, 39, 40 moisture content evaluation, 50–1 MOSFET sensors, 124–5 muguets, 16–17 multivariate analysis, 91–2, 159 autoscaling, 95–6 classification models, 99–101, 106–9 data examination, 103 data tabulation, 102–3 descriptive tests, 187 experimental design, 94 exploratory analysis, 103 general Procrustes analysis, 188 hierarchical cluster analysis (HCA), 97–8, 104–5 k-nearest-neighbor, 99–100 normalization, 95 notation, 92–4 preprocessing, 94–6 principal component regression (PCR), 101–2 principal components analysis (PCA), 98–9, 104–6, 128–9, 164–6 soft independent modeling of class analogy (SIMCA), 100–1 muscone, 14 musks, 14–15 natural aromas, 11–14 natural and artificial (N and A) flavors, 209–10 natural flavors, 210 natural products, 207, 216–17 nature-identical compounds, 2–10, 207, 210 neral, INDEX normalization, 95 noses, 112–13 see also electronic noses octanol, 63 octanol-water partition coefficient, 24–5 1-octen-3-ol, odor assessors selection and training, 70–1 sensory vocabulary, 71–2 see also olfactometry; sensory panels odor unit, 133–4 olfactometry aroma description matching, 85 charm analysis, 74–5 detection frequency method, 79–81 dilution analysis, 73–5 equipment, 72–3 mass spectrometry, 85–6 odor assessors see odor assessors OSME, 76–8 posterior intensity method, 82–3 practical considerations, 73 sample introduction, 83–4 standards, 86 time intensity analysis, 76–9 see also electronic noses; odor assessors; sensory panels optical rotation, 51–2 organic foods, 210 OSME, 76–8 osmeogram, 77 packaging, 176 paired comparison testing, 184–5 partial anosmia, 177 partial least squares regression (PLS), 131–2, 150–1, 159 pattern recognition, 97 patulin, 215 PCA see principal components analysis PCR, 101–2 PDMS see polydimethylsiloxane perfumery, 138–9 home cleaning products, 141–4 pesticides, 214–15 pharmaceuticals, 137 photoionization detectors, 126 227 INDEX plant extracts, 21 PLOT column, 60 polyacrylate, 28 polydimethylsiloxane (PDMS), 23–4, 24–5 foam extraction, 36, 37 polyethylene glycol, 29 positive lists, 204–5 preference tests, 188–9 pregnant women, 195 principal component regression (PCR), 101–2 principal components analysis (PCA), 98–9, 128–9, 142, 144 coffee samples, 164–6 exploratory, 104–6 flavors, 148 product acceptability, 175–6 product development, 176 product labeling, 206–8 product reformulation, 175 propyl sulfides, purge and trap see dynamic headspace extraction pycnometer, 52 pyrazines, 7–8 quality control, 137, 168–9, 175 quantitative descriptive analysis (QDA), 187 quartz microbalance sensors, 124 radar plot, 128 radioisotopes, 218–19 ranking tests, 185–6, 189 raspberry flavor, 150–1 refractive index, 52–4 regulations fair trade, 208–9 flavor types, 209–10 forbidden substances, 213–16 genetic modification, 219 governing authorities, 211–12 history, 202–4 other compliance requirements, 219–20 product labeling, 206–8 safety, 204–6 retention index, 56, 63–4 rhamnose, 14 rose bud ester, SAFE, 41, 42 safety regulations, 204–6 sample preparation for gas chromatographic techniques microvial, 36–9 olfactometry, 83–4 PDMS foam, 36 solid phase microextraction, 27–33 solvent extraction, 39–42 static headspace extraction, 25–6 stir bar sorptive extraction, 33–6 for sensory panel analysis, 191–2, 196 selective ion monitoring (SIM), 65–6 sensory analysis, 47, 189–90 advantages, 197–8 affective tests, 188–9 applications, 174–5 descriptive tests, 186–8 difference tests see difference tests experimental design, 189–92 flavor perception, 177–8 overall difference tests, 179–80 presentation of results, 196–7 purpose, 174–7 sample preparation, 191–2 variability elimination, 192–3 see also olfactometry; sensory panels sensory panels, 113–14, 176 conducting, 195–6 consumer, 194–5 electronic noses and, 148 results expression, 196–7 trained, 193–4 shelf life, 177 simple difference test, 182–4 Sinclair, Upton, 203 smell (human sense), 112–13 SNIF-NMR, 219 sniffers see odor assessors soft chemistry, 14 soft independent modeling of class analogy (SIMCA), 100–1, 107–8, 130–1 solid phase dynamic extraction (SPDE), 119 228 solid phase microextraction (SPME), 27–33, 28, 83–4, 119 advantages, 32 disadvantages, 32–3 fiber type, 29–30 injection port liner, 30–1 solvent-assisted flavor evaporation (SAFE), 41, 42 solvents, aroma extraction, 21 soxhlet extraction, 39–42, 40 specific gravity test, 52 Spectrum descriptive analysis, 187–8 Standard Reference Method, 48 static headspace extraction, 25–6, 160–1 statistical quality chart, 131 multiband, 132–3 steam-distilling, 19–20 stereochemistry, 218 Stevens’ Law, 76–7 stir bar sorptive extraction, 33–6 sucrose, measurement, 53–4 sugar content measurement, 53–4 sulfides, 9–10 surface acoustic wave detectors, 124–5 synthetic flavors, 10–11 INDEX tandem mass spectrometry, 65–6 TDAS-2000, 120 Tenax, 26, 27 terpenoids, 2–3 thermodesorption sampling, 119–20 tonalid, 14, 15 toxins, environmental, 215–16 transfer of calibration (TOC), 161–2 triangle test, 180–2 Trimofix, 17 tropathiane, 10 TTF natural flavors, 210 turbidity evaluation, 49 Twister, 33, 34–5 two out of five test, 180–1 niversity of Georgia, 194 valeric acid, vanilla flavor, 217 vanillin, 5, 13, 66, 66, 203 variable selection, 95 viscosity, 54 water activity evaluation, whiskey, 166–8 woodies, 17 49–50 .. .Practical Analysis of Flavor and Fragrance Materials Practical Analysis of Flavor and Fragrance Materials Edited by Kevin Goodner and Russell Rouseff A John Wiley... extraction and fast, relatively simple Practical Analysis of Flavor and Fragrance Materials, First Edition Edited by Kevin Goodner and Russell Rouseff © 2011 Blackwell Publishing Ltd Published 2011 by. 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First Edition Edited by Kevin Goodner and Russell Rouseff © 2011 Blackwell Publishing Ltd Published 2011 by Blackwell Publishing Ltd 2 PRACTICAL ANALYSIS OF FLAVOR AND FRAGRANCE MATERIALS human