Perfumes☆

Alberto Chisvert, University of Valencia, Valencia, Spain

Marina López-Nogueroles, University of Valencia, Valencia, Spain; Unidad Analítica, Instituto de Investigación Sanitaria La Fe (IIS La Fe),
Valencia, Spain
Amparo Salvador, University of Valencia, Valencia, Spain
© 2018 Elsevier Inc. All rights reserved.

Introduction 1
Perfumed Products 1
Perfume Raw Materials 1
Legislation and Safety 3
Perfume Analysis 4
References 6
Further Reading 6

Introduction

The use of perfumes goes back from antiquity to our days, in which they are everyday products. Nowadays many people feel the
need to wear a perfume or to perfume their home in order to feel good. This can be explained by the relation between scent and
emotion as well as between scent and memory. Moreover, studies have shown that some scents can modulate different physiolog-
ical pathways.1
The word perfume can be defined as a preparation that releases a pleasant smell and that is usually a liquid based on natural or
synthetic ingredients. The word fragrance is usually reserved to refer to a perfuming ingredient used either in a perfume or in a
perfumed formulation.
Perfumes consist of a mixture of aromatic chemicals and essential oils. Up until the 19th century they were usually composed of
natural aromatic oils but nowadays, most perfumes are synthetic and may contain many fragrance chemicals.
This article deals with the different types of perfumed products, either perfumes or perfumed formulations containing fragrances,
their legislation and safety aspects and the analytical methods for fragrance chemicals determination.

Perfumed Products

More than 2500 fragrances are used today in different types of products such as cosmetics and household products (Fig. 1).
Perfumes used in cosmetics are complex mixtures of hundreds of ingredients (fragrances and other components), which together
give the product its unique scent. They are used in practically all cosmetics, not only in perfumes (also named fine fragrances) but
also in other perfumed cosmetic products such as: general toilet products (bath gels, soaps, deodorants, etc.), skin care products
(face and body care products), sunscreens and related products (sun products, bronzing products, whitening products), hair care
and related products (shampoos, conditioners, hairsprays, shaving creams, etc.) or decorative cosmetics (make-up products, tinted
based products, etc.).
Fine fragrances are hydroethanolic solutions prepared from aromatic chemicals and essential oils with the main function of
perfuming us, providing a pleasing or attractive scent. Fine fragrances are named differently according to the perfume content
Extract, Parfum, Eau de Parfum, Eau de Toilette, Eau Fraiche, Eau de Cologne, Cologne, Baby Cologne (Fig. 2). This last one has a
special formulation, with a very low content of perfume and some moisturizing ingredients added to give softness to baby skin.
Other cosmetics, such as aftershave lotions also contain a high content of perfumes, in the range of 0.5%–2%.
A wide range of perfumes, predominantly based on synthetic ingredients, are used in household products. For example,
perfumes are used in air fresheners to provide a desired pleasing smell to our homes, work or public spaces. They are also used
to make many cleaners, such as softeners and laundry detergents, dishwashing products, floor cleaning products, furniture polish or
bathroom cleaners, smell more pleasant than their own chemical ingredients.
Special problems can occur in the formulation of cosmetics or household products containing perfumes, since odors may change
or become unstable in the new medium or the perfume may change the physical properties of the product, etc. Therefore, the
perfume must be individually formulated for each product.

☆
Change History: March 2018. Alberto Chisvert, Marina López-Nogueroles, and Amparo Salvador Updated the text, Fig. 1 and Fig. 2 of the 2nd Edition, deleted
Table 2, Fig. 3 and Fig. 4 of the 2nd Edition, added abstract, a new figure (Fig. 3 of the 3rd Edition), a new table (Table 2 of the 3rd Edition), a list of Relevant
Websites, a list of References, and a list of Further Reading.

Encyclopedia of Analytical Science, 3rd Edition https://doi.org/10.1016/B978-0-12-409547-2.14037-5 1

2 Perfumes

PERFUMED PRODUCTS

COSMETIC HOUSEHOLD
PRODUCTS PRODUCTS

Perfumes or
Air fresheners
fine fragrances

Perfumed Other perfumed

cosmetic products household products
– General toilet products – Laundry products
– Skin care products
– Home cleaners
– Sunscreens and related
products
– Hair care and related products
– Decorative cosmetics

Fig. 1 Types of products usually containing perfumes.

Parfum 20-40 %

Eau de Parfum 15-20 %

Eau de Toilet 5-15 %

Eau Fraiche 4-5 %

Eau de Cologne 3-4 %

Cologne 2-3 %

Baby cologne 1-2 %

Fig. 2 Approximate concentration of perfume in different types of fine fragrances.

Perfume Raw Materials

Raw materials used as perfumes include natural and synthetic materials.

Natural ingredients are obtained from plant or animal products.
Plant based ingredients are obtained from different parts of the plants for example, flowers (jasmine, rose, gardenia), fruits
(lemon, orange), roots (vetiver, cistus), leaves (violeta, patchuli), woods (vetiver, sandalwood, cedarwood), barks (cinnamon,
nutmeg), resins (benjui, tolu, galbano), seeds (angelica, apio) or whole plants (lavander, geranio).
These plant based ingredients are obtained through different physical processes, such as:

▪ Hydrodistillation. It is used to obtain essential oils (lavender, rose) from plants by boiling them in water, evaporation of the
essential oils and condensation.
▪ Solvent extraction. It is used for delicate flowers (e.g., jasmine) as it avoids the use of high temperatures. The concentrated flower
oil is called absolute.
▪ Sohxlet extraction. It can facilitate the extraction of essential oils in some cases.
▪ Supercritical fluid extraction. It allows faster and greener extraction procedures, avoiding the use of organic solvents.
▪ Enfleurage. In this procedure, petals are placed between layers of purified animal fat, which become saturated with flower oil,
alcohol is then used to obtain the absolute. This technique is carried out in cold, is very costly and rarely used today.
▪ Maceration. It is similar to enfleurage. The flowers are steeped into vats of oil until the scented parts dissolve. It is a time
consuming procedure. The oil may be heated to speed up the process.
 Perfumes 3

▪ Expression. It is used to obtain essential oils from fresh fruit peels (lemon, orange). It can be done either by using traditional
procedure of pressing with sponges or by mechanical maceration, as there is no heat involved. The oil smells very close to the
original plant.
Apart from these techniques there are various other operations such as rectification, fractional distillation, terpenes removal,
decolourisation, etc., which improve and refine the numerous raw materials used for the blending and making of perfumes.
Some animal secretions contain odoriferous substances that increase the diffusion and qualities of perfumes or can act as
fixatives, preventing the rapid evaporation of the most volatile ingredients of the perfume. Some examples are: ambergris (from the
sperm whale), castor or castoreo (from the beaver), civet (from the civet cat) and musk (from the musk deer). Natural ingredients
coming from animal products are usually extracted using the alcohol maceration procedure.
Diverse types of synthetic ingredients are also used in perfumes. As the components of a great number of natural perfumes have
been determined, they can be chemically synthetized to imitate the natural perfumes. However, artificial fragrances that do not try to
imitate nature are also used to create new and original perfumes.
In a chemical perspective, the compounds which perfume manufacturers use to achieve the desired smells are very different
between them and can be grouped into different families according to their chemical structure. Table 1 shows some examples of
these compounds.
The main advantages of using synthetic ingredients is that costs are lower than those for natural ingredients and ease and
reproducibility in its obtaining. Natural ingredients depend on problems such as poor crop quality, lack of supplies or difficulties
using animal extracts, etc. In other words, artificial products cost less and suffer fewer market variations.
The selection of the raw materials depends on many different questions such as: desired odor, target users, types of applications,
stability of the finished product, range of price fixed for the product, etc.

Legislation and Safety

Toxicological aspects and health risks associated with perfumes have been widely studied. Most of the safety testing performed has
focused on the dermatological effects of fragrance chemicals as they are the most frequent. The effects on the respiratory system, the
brain or other organs of the body have been less studied. Fragrance chemicals can be inhaled through the nose, ingested through the
mouth, and absorbed through the skin. Once in the body they reach the bloodstream and are transported throughout the body.
Individual sensitivity to the effects of fragrance chemicals vary widely from no effect at all to severe reactions. Different symptoms
have been found in different people: skin sensitivity, rashes, dermatitis, coughing, asthma attacks, migraine, etc. The incidence of
these effects can be studied in relation to the method of application: leave-on skin, rinse-off skin, nonskin contact, household
products, etc. The Scientific Committee on Consumer Safety (SCCS) published an opinion on fragrance allergens in cosmetic
products in 2012 (SCCS/1459/11), including a large number of references on the published literature dealing with toxicological
effects of the different fragrances. Other interesting publications on this topic can be found in later publications.2–4
Safety legislation on perfumes are adopted in different countries. European Union (EU) (Regulation EC 1223/2009) and United
States (US) (FDA, Code of Federal Regulations, Title 21) legislations stipulate that cosmetic ingredients must be listed on the label in
order of predominance. However, perfumes are one of the exceptions as their composition are considered trade secrets and thus, the
list of ingredients is not required. In this sense, legislations allow to include just the word “parfum” or “aroma” (EU) or the words
“fragrance” or “flavor” (US) on the label of those products where ingredients have been added to give an odor. So, this word on the
label may represent many ingredients.
Since perfume raw materials do not have to be declared on the label, it is impossible to know exactly which substances are
present in the product. This makes difficult to ascertain if a potentially problematic substance is present in any given product.
Fortunately, since 2003 (Directive 2003/15/EC), EU legislation (Regulation EC 1223/2009) require manufactures to declare on the
label the presence of 26 fragrance substances which have proven to be allergenic. Table 2 shows the 26 substances potentially

Table 1 Illustrative example of compounds that could be present in perfume composition

ordered according to its chemical structure

Group Compound

Alcohol b-Citronellol, eugenol, geraniol, linalool, menthol

Aldehyde Benzaldehyde, citral, hexyl cinnamal, anisaldehyde, vanillin
Hydrocarbon Nonane, tetradecane, camphene
Terpene Limonene, myrcene, a-pinene
Sesquirtepene Caryophyllene
Ester Methyl salicylate, menthyl lactate, terpineol acetate
Ether Anethole, eucalyptol
Ketone Cyclohexanone, camphor
Lactone Coumarin, 6-methyl-coumarin, g-undecalactone
4 Perfumes

Table 2 Potentially allergenic fragrances that have to be declared on the label of cosmetic
formulations according to the EU legislation

Common name CAS number

Amyl cinnamal 122-40-7

Amylcinnamyl alcohol 101-85-9
Anisyl alcohol 105-13-5
Benzyl alcohol 100-51-6
Benzyl benzoate 120-51-4
Benzyl cinnamate 103-41-3
Benzyl salicylate 118-58-1
Cinnamal 104-55-2
Cinnamyl alcohol 104-54-1
Citral 5392-40-5
Citronellol 106-22-9
Coumarin 91-64-5
Eugenol 97-53-0
Farnesol 4602-84-0
Geraniol 106-24-1
Hexyl cinnamaldehyde 101-86-0
Hydroxycitronellal 107-75-5
Hydroxymethylpentyl-cyclohexenecarboxaldehyde 31906-04-4
Isoeugenol 97-54-1
Lilial 80-54-6
D-Limonene 5989-27-5
Linalool 78-70-6
Methyl heptine carbonate 111-12-6
3-Methyl-4-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3-buten-2-one 127-51-5
Oak moss 90028-68-5
Tree moss 90028-67-4

allergenic which presence in cosmetic formulations has to be advertised according to the EU legislation. From these 26 substances,
two are natural extracts (oak moss and tree moss) and the rest are volatile chemicals.
In addition to these 26 substances, there are others that have also been established as contact allergens in humans, although with
a lower incidence. The EU plans to add an extra list of allergens which presence will also have to be mentioned on the label and to
ban some others allergens (SCCS/1459/11). This initiative and other similar ones in other countries should contribute to have
better informed consumers.
Furthermore, EU legislation on cosmetic products forbids the use of more than 30 perfuming substances or extracts, such as
some synthetic musk. Although US legislation does not forbid any fragrance ingredient, there are some references about the need to
study their possible allergenic effects.
Different organizations such as the “Research Institute for Fragrance Materials” (RIFM) (RIFM website) or the “International
Fragrance Association” (IFRA) (IFRA website) were founded by the industry and have an important role in consumer protection.
They carry out research on ingredients used in perfumes, in order to ensure the safety of perfumery materials. An available database
on toxicology for flavor and fragrance materials is maintained by RIFM, classifying more than 4500 materials.
It is worth mentioning that terms such as “unscented” or “fragrance free” have no legal definitions as there is not a declared list of
fragrance substances. In the same way, the term “hypoallergenic” is sometimes used in some cosmetics with marketing purposes.
Perfumes are the main responsible for producing allergies, although some preservatives, dyes, UV filters, surfactants or solvents
can also be allergenic. Hypoallergenic cosmetics are products that manufacturers claim to produce fewer allergic reactions than other
cosmetic products. However the term has no legal definition and consequently the use of a hypoallergenic product does not
guarantee immunity to any allergic reaction.

Perfume Analysis

Analytical control of the fragrances in perfumed formulations ensures that the final product has the desired smell in the desired
amount.
Special interest have the fragrances that must be declared on the label because although they are permitted, they could cause
health problems. For instance, the above mentioned 26 potentially allergenic substances according to the EU legislation (see
Table 2). Also, synthetic musk, such as musk ambrette, musk tibetene and musk moskene, are analytes of interest as they are
prohibited because they have been linked to dermatitis and possible carcinogenic effects and endocrine dysfunctions. Other musks,
 Perfumes 5

such as musk xylene and musk ketone, are permitted with restrictions. Different publications focused on analytical methods for
determining all of these and other substances have been recently reviewed.5
Sample preparation not only depends on the type of sample but also on the analytical technique used for analysis. Sometimes,
sample pretreatment is not necessary due to the simplicity of some of these samples (such as fine fragrances), and they can be
directly (or after dilution in organic solvents) analyzed by chromatographic techniques. However, in most cases extraction
techniques, like liquid–liquid extraction, solid phase extraction in combination with liquid–liquid, online clean-up with thermal
desorption, matrix solid-phase dispersion, pressurized liquid extraction, ultrasound assisted microextraction, etc. are used not only
for achieving the isolation of the target compounds from the matrix but also to produce a preconcentration of the analytes.
Physical (i.e., refractive index, density or optical rotation) or simple chemical test (acidimetry, saponification, carbonyl index)
are useful for checking the raw materials or the final product, but they do not offer an actual determination of the compounds that
are present in an unknown perfume. Taking into account that many compounds form part of a perfume, the determination of the
composition of an unknown perfume is not an easy task. As an example, fine fragrances can contain over 50 ingredients and,
although the expertise of the perfumers is sometimes useful, it is not enough to identify (and much less to quantify) all the
compounds involved. In this sense, powerful analytical techniques are required. Separation techniques like chromatographic and
related techniques are the most suitable for individual separation and determination of these compounds.
Bearing in mind that the compounds forming the perfume have a low boiling point, gas chromatography (GC) has been the
most used technique in the perfume industry. Due to the volatile nature of the compounds present in a perfume, the head-space
(HS) sampling mode is also commonly employed to introduce the compounds to be determined in the GC system. In this sense, the
perfume, after appropriate sample pretreatment, can be analyzed by GC with a flame ionization detector (FID) and the Kovats Index
(KI) for each peak can be experimentally determined. The identification is carried out by comparing the experimentally determined
KI with those values kept in databases, although sometimes a composition is not completely resolved because there can be peaks
with very close KI. Moreover, although the databases update quite often, new ingredients can be present, and the software library
will not recognize these novel compounds. In these cases, a mass spectrometry (MS) detector coupled to GC, instead of a FID, plays
a crucial role, since a chemical structure can be elucidated by studying the mass spectra of the compound.6 The use of GC–MS has
been satisfactorily applied to the analysis of different cosmetic products such as shampoos, creams, lotions, lipsticks and face
powder, and moreover to fine fragrances. In this sense, current databases, contain the mass spectra of compounds besides the KI,
becoming GC–MS in a powerful analytical technique. The internal standard calibration method is often used for quantitative
requirements. Selected ion-monitoring (SIM) mode can be very useful, especially in trace analysis, making it more sensitive and
selective.

Fig. 3 (A) Chromatogram of a lab-made cosmetic formulation containing potentially allergenic fragrance ingredients at 10 ppm. The chromatogram was recorded
in SIM. (B) Chromatogram recorded in full scan mode of a commercial cosmetic sample containing seven fragrance ingredients above labeling limit. 1. Limonene, 2.
Benzyl alcohol, 3. Linalool, 4. 1-Fluoronaphthalene (IS), 5. Methyl 2-octynoate, 6. Citronellol, 7. Citral 1, 8. Geraniol, 9. Citral 2, 10. Cinnamal, 11. Anisyl alcohol, 12.
Hydroxycitronellal, 13. Cinnamyl alcohol, 14. Methyl decanoate (IS), 15. Eugenol, 16. Coumarin, 17. Isoeugenol, 18. a-Isomethyl ionone, 19. Methyl laurate (IS), 20.
Lilial, 21. Amyl cinnamal, 22. Lyral 1, 23. Lyral 2, 24. Amyl cinnamyl alcohol, 25. Farnesol 1, 26. Farnesol 2, 27. Hexyl cinnamal, 28. Benzyl benzoate, 29. Methyl
pentadecanoate (IS), 30. Benzyl salicylate, 31. Benzyl cinnamate. Reprinted with permission from Desmedt, B.; Canfyn, M.; Pype, M.; Baudewyns, S.; Hanot, V.;
Courselle, P.; De Beer, J.O.; Rogiers, V.; De Paepe, K.; Deconinck, E. (2015). HS-GC–MS method for the analysis of fragrance allergens in complex cosmetic
matrices. Talanta 131, 444–451, Elsevier.
6 Perfumes

Fig. 3 shows the obtained chromatograms for two cosmetic formulations containing potentially allergenic fragrance ingredients
by using a Headspace Gas Chromatography-Mass Spectrometry (HS-GC–MS) method, after extraction with hexane. The chromato-
gram (SIM mode) corresponding to a lab-made sample shows 31 peaks Fig. 3 (A): 27 corresponds to 24 allergenic fragrances (3 of
them were each detected as 2 separate single peaks) and 4 to the internal standards (IS). Fig. 3 (B) shows the chromatogram (full
scan mode) of a commercial sample containing 7 allergenic fragrances without any interferences from other components.
In a recent publication the determination not only of the abovementioned potentially allergens but also of other 28 and their
isomers by comprehensive two-dimensional GC coupled with both MS and FID in parallel is proposed.7
Other chromatographic techniques, like liquid chromatography (LC) and thin layer chromatography (TLC) have been less used
in perfume analysis. Different compounds such as benzylideneacetone, cinnamyl alcohol, isoeugenol, cynnamyl anthranilate,
cynnamaldehyde, bergapten, citropten, coumarin, 6-methyl coumarine or safrole have been determined by using either LC or TLC.
Many of the compounds present in perfumes are chiral, and sometimes the chiral ratio of a mixture can affect the odor quality of
the perfume, since the olfactory receptor in the human nose is protein based and thus enantiospecific. Chiral columns can achieve
the separation and quantification of the different enantiomers of a target compound.8,9
Moreover, electronic noses also have an increasingly prominent role in the field of perfume analysis. These instruments are
devices composed of an array of nonselective gas sensors that can act in a similar manner to real biological noses. The characteristic
signal patterns can be conveniently processed by using multivariate data analysis, partial least squares, artificial neuronal network,
etc. and they can be used for the identification of different compounds and the differentiation between authentic and fake fine
fragrances.10

References

1. Angelucci, F. L.; Silva, V. V.; Dal Pizzol, C.; Spir, L. G.; Praes, C. E. O.; Maibach, H. Physiological Effect of Olfactory Stimuli Inhalation in Humans: An Overview. Int. J. Cosmet. Sci.
2014, 36, 117–123.
2. Basketter, D.; Kimber, I. Fragrance Sensitisers: Is Inhalation an Allergy Risk?Regul. Toxicol. Pharmacol. 2015, 73, 897–902.
3. Frosch, P.; Johansen, J. D.; Schuttelaar, M. L.; Silvestre, J. F.; Sanchez-Perez, J.; Weisshaar, E.; Uter, W. Patch Test Results With Fragrance Markers of the Baseline Series—
Analysis of the European Surveillance System on Contact Allergies (ESSCA) Network 2009–2012 on Behalf of the ESSCA Network. Contact Derm. 2015, 73, 1631–1671.
4. Uter, W.; Johansen, J. D.; Borje, A.; Karlberg, A. T.; Liden, C.; Rastogi, S.; Roberts, D.; White, I. R. Categorization of Fragrance Contact Allergens for Prioritization of Preventive
Measures: Clinical and Experimental Data and Consideration of Structure–Activity Relationships. Contact Derm. 2013, 69, 196–230.
5. Chisvert, A.; López-Nogueroles, M.; Salvador, A. Perfumes in Cosmetics: Regulatory Aspects and Analytical Methods. In Analysis of Cosmetic Products, 2nd ed.; Salvador, A.,
Chisvert, A., Eds.; Elsevier: Amsterdam, 2018; Chapter 10.
6. Van Asten, A. The importance of GC and GC-MS in Perfume Analysis. Trends Anal. Chem. 2002, 21, 698–708.
7. Belhassen, E.; Bressanello, D.; Merle, P.; Raynaud, E.; Bicchi, C.; Chaintreau, A.; Cordero, C. Routine Quantification of 54 Allergens in Fragrances Using Comprehensive Two-
Dimensional Gas Chromatography-Quadrupole Mass Spectrometry With Dual Parallel Secondary Columns. Part I: Method Development. Flavour Fragr. J. 2018, ;63–74.
8. Bicchi, C.; Cagliero, C.; Rubiolo, P. New Trends in the Analysis of the Volatile Fraction of Matrices of Vegetable Origin: A Short Overview. A Review. Flavour Fragr. J. 2011,
321–325.
9. Cagliero, C.; Sgorbini, B.; Cordero, C.; Liberto, E.; Rubiolo, P.; Bicchi, C. Enantioselectives Gas Chromatography with Cyclodextrin in Odorant Analysis. In Springer Handbook of
Odor; Buettner, A., Ed.; Springer Handbooks, 2017.
10. Gebicki, J.; Szulczynski, B.; Kaminski, M. Determination of Authenticity of Brand Perfume Using Electronic Nose Prototypes. Meas. Sci. Technol. 2015, 26, 125103.

Further Reading
Chaintreau, A. Analytical Methods to Determine Potentially Allergenic Fragrance-Related Substances in Cosmetics. In Analysis of Cosmetic Products, 1st ed.; Salvador, A., Chisvert, A.,
Eds.; Elsevier: Amsterdam, 2007; Chapter 6, Section 2.
Chisvert, A.; Salvador, A. Perfumes in Cosmetics. Regulatory Aspects and Analytical Methods for Fragrance Ingredients and Other Related Chemical in Cosmetics. In Analysis of
Cosmetic Products, 1st ed.; Salvador, A., Chisvert, A., Eds.; Elsevier: Amsterdam, 2007; Chapter 6, Section 1.
Negri, R. M. Electronic noses in Perfume Analysis. In Analysis of Cosmetic Products, 1st ed.; Salvador, A., Chisvert, A., Eds.; Elsevier: Amsterdam, 2007; Chapter 6, Section 3.
Sell, C., Ed.; The Chemistry of Fragrances. From Perfumer to Consumer, The Royal Society of Chemistry: Cambridge, 2006.

Relevant Websites

http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/cfrsearch.cfm—FDA (U.S. Food and Drug Administration), Code of Federal Regulations, Title 21, Parts 70-82 for Colorants;
Parts 330-360 for OTC drugs; Parts 700-740 for Cosmetics.
http://ec.europa.eu/health/scientific_committees/consumer_safety/docs/sccs_o_102.pdf—SCCS/1459/11, Opinion on Fragrance allergens in cosmetic products.
http://eur-lex.europa.eu/eli/reg/2009/1223/2016-08-12—Regulation (EC) 1223/2009 of the European Parliament and the Council of 30 November 2009 on cosmetic products.
http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri¼OJ:L:2003:066:0026:0035:en:PDF—Directive 2003/15/EC of the European Parliament and of the Council of 27 February
2003, amending Council Directive 76/768/EEC on the approximation of the laws of the Member States relating to cosmetic products.
http://www.ifraorg.org/en-us/ingredients—IFRA (International Fragance Association) website.
http://www.rifm.org—RIFM (Research Institute for Fragrance Materials), website.