Received: 27 May 2017 Revised: 3 August 2017 Accepted: 17 August 2017

DOI: 10.1002/ffj.3418

REVIEW

The essential oil of patchouli, Pogostemon cablin: A review*

Teris A. van Beek1 | Daniel Joulain2

1
Laboratory of Organic Chemistry,
Wageningen University, Stippeneng 4, 6708
WE Wageningen, The Netherlands
2
SCBZ Conseil, Les Micocouliers ‐ F3, 99
avenue Sidi Brahim, 06130 Grasse, France
Correspondence
Teris A. van Beek, Laboratory of Organic
Chemistry, Wageningen University,
Stippeneng 4, 6708 WE Wageningen, The
Netherlands. Tel. + 31 317 482376;
Fax + 31 317 484914;
Email: teris.vanbeek@wur.nl
Abstract
The leaves of Pogostemon cablin (Blanco) Benth. (Lamiaceae) are the source of patchouli
essential oil, which is – with an annual production of about 1300 tonnes – an important
and unique commodity in the fragrance industry. All the literature pertaining to patchouli
was critically reviewed with an emphasis on the qualitative and quantitative chemical analysis
of the oil but also harvesting, fermentation, drying, distillation, used analytical techniques,
sensory aspects including molecules responsible for the odour, adulteration and toxicological
aspects, i.e., skin sensitisation, are discussed. In total 72 constituents have been convincingly
identified in the oil and another 58 tentatively. The main constituent is the sesquiterpene
patchoulol. For this review over 600 papers were consulted and in the supplementary
information all patchouli‐related references not relevant enough to be cited in the paper itself
are listed.

KEY W ORDS

adulteration, composition, odour, patchoulol, steam distillation, toxicology

1 | I N T RO D U CT I O N constituents,15 and a short monograph containing information on

Patchouli essential oil (PEO) is obtained by steam distillation or
hydrodistillation of the dried leaves of Pogostemon cablin (Blanco)
Benth. (Lamiaceae). It has a unique woody odour and is an indispensable
and difficult to replace constituent of many women's and men's
fragrances as well as cosmetics in general.6 It has been stated that “this
oil is one of the most important materials available to the perfumer.”7
Approximately 90% of today's global production of 1200‐1300 metric
tonnes per annum is realised in Indonesia.8-11 Swamy et al. mention a
higher figure of about 2000 tonnes.12 In terms of bulk, it is approxi-
mately the tenth most important essential oil.8 The current price level
is US $ 61‐63 per kg for good quality oil11 but has been as high as $
150/kg.13 In terms of turnover PEO belongs to the 15 most important
essential oils,14 and we estimate the current annual sales at approxi-
mately $ 75 million. Patchouli leaves entered the European market in
the first half of the 19th century and the odour soon became popular.
The essential oil is already mentioned in the first book on plant
constituents and odour was published as early as 1875.16 More exten-
sive monographs were published near the end of the 19th century17,18
and PEO constituents like patchoulol and patchoulene already occur in
Beilstein's Handbook of Organic Chemistry.19
With regard to the Chemical Abstracts Service (CAS) Registry
Number (RN) assigned to the essential oil, the situation is somewhat
confusing. In our view the CAS RN, which corresponds best to the
essential oil is [8014‐09‐3] (Oils, patchouli), the definition of which is:
“Extractives and their physically modified derivatives. Pogostemon
cablin (Pogostemon patchouli), Labiatae” with as other names: Oil of
Patchouli, Patchouli oil. In addition to this RN, there is also [84238‐
39‐1] (Patchouli, extract) with as definition: “Extractives and their
physically modified derivatives such as tinctures, concretes, absolutes,
essential oils, oleoresins, terpenes, terpene‐free fractions, distillates,
residues, etc., obtained from Pogostemon cablin (Pogostemon patchouli),
Labiatae”. This is clearly a much wider definition also encompassing
solvent extracts and supercritical fluid extracts. However, it does not
exclude the essential oil. Still other CAS RNs, like [1442462‐94‐3],

*Part 6 of 7 in a series on woody fragrance raw materials. For parts 1‐5 see [918959‐91‐8], [94334‐20‐0], [91770‐50‐2], [91770‐49‐9], [91079‐
references 1–5.1–5 37‐7], [73049‐67‐9], [68917‐23‐7], and [73049‐85‐1] refer to specific

--------------------------------------------------------------------------------------------------------------------------------
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided
the original work is properly cited.
© 2017 The Authors. Flavour and Fragrance Journal Published by John Wiley & Sons, Ltd.

6 wileyonlinelibrary.com/journal/ffj Flavour Fragr J. 2018;33:6–51.

 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 7

modified extracts, e.g., acetylated or oxidised extracts, and are of little too. All papers were obtained in complete form and are not cited from
importance for this review. [1450625‐49‐6] refers to a biotechnologi- abstracts only. In some cases, external experts or authors of published
cally produced extract, which is reminiscent of patchouli but is not patchouli papers were consulted for advice. Reliable references
derived from P. cablin. In the USA, PEO has been approved by the Food relevant to the fertilisation, harvesting, drying, fermentation and
and Drug Administration under paragraph 21 CFR 172.510,20 and has distillation of patchouli leaves, identification and chemical analysis of
21
the GRAS (“generally recognized as safe”) status under nr 2838. It volatile constituents of patchouli, and the odour, adulteration and
occurs in the list of the European Chemical Agency (ECHA) under EC toxicology of the oil finally made it into this review. Some analytical
list nr 616‐944‐7,22 and in the Chinese positive list of ingredients used papers were deemed less reliable and were excluded. This review
in cosmetics under nr 02634.23 excludes a discussion of the history, botany, ecology, breeding, cultiva-
Over the years many reviews have appeared on Pogostemon cablin, tion, biotechnology, pharmacology, medical and food uses, economy
its constituents and their uses. They are summarised in Table 1. In spite and biosynthesis of constituents or the oil itself. For these topics, the
of the large number of reviews, a comprehensive and critical review of reader is referred to one of the many reviews cited in Table 1. All
the chemical composition of PEO and its quality control, adulteration, encountered patchouli references, which are not cited in this review,
odour and possible skin sensitisation is still lacking, i.e., a review for are listed in the Supplementary Information (SI).
the fragrance industry similar to the ones, which appeared earlier in
this journal on plants containing woody fragrances.1-5 This review aims
at filling this gap.
2 | I N F L U E NC E OF F ER T I L I S A T I O N ,
H A RV E S T I N G T I M E , D R Y I NG ,
TABLE 1 Overview of earlier reviews on P. cablin, its constituents or F ER M E N T A TI O N A N D D I S T I L LA T I O N
its uses TE C HN I Q U E ON ES SE N TI A L OI L CO N TE N T
Year of
A N D C O M P OS I T I O N
Main topics of review publication References
24-27
broad, i.e., history, botany, 1911, 1921, 2.1 | Average Essential Oil Content
cultivation, distillation, annual 1931, 1949
production and prices The average oil percentage in “dried” P. cablin leaves as
28-32
cultivation of patchouli 1968, 1986, reported in 72 references was calculated as 2.6% (ranges: 0.54 –
1988, 1994,
2006
5.2%).12,26-28,55,58-121 However in view of the discussion below, this

trade of PEO 1982 33,34 figure is not precise as it is dependent on the exact plant material used

chemistry 1984, 1995 35,36 (cultivar, age, soil and growth conditions), percentage of stems as well as

mini reviews on chemistry 1976‐2014 37-43 foreign materials (soil, other plant species like weeds122), drying and/or

agrotechnology 1995 44 fermentation, storage, final percentage of water and the distillation

uses in foods 1996, 2016 45,46 technique, conditions and duration. There is no standardisation so it is

all aspects except for chemical 1997 47 well possible that the percentage PEO of two batches of identical plant
constituents material could differ more than a factor of two. A few studies reported
48
botanical and biological aspects 1999 the oil percentage in fresh leaves on a fresh weight basis, a realistic
general review 2005, 2015 13,49
value being ~0.3%.123 Mahanta reported a seemingly high value of
role of P. cablin as herbal medicine 2007 50
0.7%.122 Twigs or stems contain less oil than leaves.65,75 Husain stated
in China
that an oil content of 2.5% is satisfactory for commercial purposes.30
origin of the name “patchouli” 2010 51
Weiss cites Indonesian sources, who claim 5.8%, or even 6‐8% oil
52
odour 2012
content, but this seems optimistic in view of all the reports above.47
53,54
phytochemistry and pharmacology 2013, 2015
55
production and application 2013
cultural practices, harvesting, 2016 56 2.2 | Effects on Oil Composition
distillation and diseases
57
In view of the difficulties associated with the chemical analysis of PEO
botany, agrotechnology and 2016
biotechnology (see § 3.2), the natural variation of patchouli leaves and the lack of
standardisation of various procedures such as fertilisation, harvesting,
drying, fermentation or distillation, in our view there are currently no
References were obtained from the authors' existing personal reliable studies on the effect of said parameters on the percentage of
literature databases on patchouli as well as a search in SciFinder individual constituents in PEO. Thus, this aspect is not further
Scholar from 1900 till October 20, 2017 with the key words: discussed.
“Pogostemon cablin”, “patchouli”, “patchouly” and “patchoulol” but
restricted to English, French and German publications in journals. Addi-
2.3 | Fertilisation and Cultivation Conditions
tionally, Miltitz Berichte and Fritzsche, Dodge and Olcott monthly bul-
letins were scanned. More references in more languages were found Bhaskar and Saha et al. compared different nitrogen treatments. The
after reading all the initially obtained papers and these were subse- oil yield in kg/ha increased with more nitrogen but an effect on the
quently downloaded and read. Several relevant patents were included oil content (in %) was less clear, with only the lowest N dose giving

8 VAN BEEK AND JOULAIN
significantly less oil.73,95,96 Also in similar later studies, no or a very lim-
ited influence of fertiliser levels on oil content was found but the yield
in kg/ha invariably increased.76,87,88,99-102,124,125 Additionally Singh
found no effect of the irrigation level on oil content or oil composi-
tion.102 No effect of potassium on the oil content was found.92
Bhaskar et al. investigated the effect of two growth regulators
(triiodobenzoic acid and kinetin) on oil content and oil yield. At the
maximum dose of both, the oil yield (in kg/ha) more than doubled
but the oil content increased only by 25%.73,103 In contrast, in another
study a 50 μM application of kinetin gave a 58% increase in % oil con-
tent on a shade dry weight basis.126 Application of benzyl adenine (BA)
gave a 50% increase in oil yield but did not influence oil content.94
Misra obtained the same result for foliar application of gibberellin.85
When patchouli plants were grown under sun and shade conditions,
84
there was no significant difference in oil content in one study, while
in two other studies unshaded plants contained more oil.28,47 When 11
different arbuscular mycorrhizal fungi were tested, one was found to
increase significantly both biomass and oil content.104
2.4 | Harvesting Time
The first harvesting usually takes place six months after planting in the
field, and then again, every three to six months for a total of two to
three years. Harvesting is best done after sunrise or before sun-
set.27,30,47,48 Tripathi and Hazarika investigated the influence of the
harvest year and the harvest within each year on the oil content. The
oil content declined from 4.1% in year 1 via 3.5% in year 2 to 2.7%
in year 3. There were no significant differences between the three har-
vests within each year.59 The same authors studied the influence of
the maturity stage. The oil content of semi‐mature leaves (4.5%) was
higher than those of vegetative (4.0%) and fully mature leaves
(3.8%).59 Blank et al. harvested four times within the first year after
planting and found average percentages of 1.73, 1.70, 2.14 and
1.79% for 7 different plants.61 Manjunatha et al. reported approxi-
mately the same oil percentage for harvests 6, 9 and 12 months after
planting. The oil yield per ha decreased sharply for these same har-
87
vests, especially from the first to the second harvest. Similar results
have been reported by Singh et al.110 In contrast, in another study both
the oil content and the oil yield per ha were much higher for the sec-
ond harvest after 11 months than for the first harvest after 8
months.97 Sarma compared the oil yield in kg/ha of harvests after 5,
8 and 11 months in two subsequent years. In both year 1 and 2, the
highest yields were noticed after 11 months and on average yields
were 40–50% higher in year 2 than in year 1. The influence of the har-
vesting time during the day (morning, mid‐day, evening) was much less
124
pronounced. In yet another study, leaves were harvested after 3, 6
68
or 9 months. Oil yields were 3.45, 3.36 and 2.38%. Unusually Thai
et al. harvested every month from the 4th month after planting
onwards.120 The oil yield increased from an initial ~2.2% to ~3.1%
for the last three harvests. As the amount of leaves in kg/ha decreased
sharply during the last two harvests, the optimum oil yield in kg/ha
rd th 120
peaked at the 3 and 4 harvest (~14 kg/ha). Overall the different
studies do not yield one conclusive picture and most likely local influ-
ences like plantlet type, soil, rain, temperature, weeding, manure,
amount of leaves, distillation parameters, etc. play an important role.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
Kongkathip et al. investigated the patchoulol content of PEOs
obtained from leaves harvested after 3, 6 or 9 months and noted a
gradual decrease as a function of leaf age.68
2.5 | Drying and Fermentation
As about 50% of the available PEO is stored in cells buried within the
leaves (see next section on distillation for more information), the distil-
lation of fresh leaves gives a low yield, also when expressed on a dry
weight basis. The best way to free the interior oil is drying and possibly
also a mild fermentation of the leaves. These processes make the cell
walls of the oil glands more permeable, making it in turn easier for
PEO to diffuse out of the leaves and be steam distilled. This was
already more than a century ago well documented by de Jong.127 He
determined the oil content of fresh leaves, leaves fermented during
several days at 52 °C and leaves dried immediately after harvesting
and found that the dried and fermented leaves gave exactly the same
yield, which was 2.5 times higher than that of distilled fresh leaves.127
Later studies confirmed this finding.65,83 During the fermentation,
there was no enzymatic action on stored non‐volatile precursors to
increase the amount of PEO127 thus disproving an earlier hypothesis.18
Similarly, Guenther stressed that proper drying is important as it makes
the membranes more permeable. However fermentation should be
avoided as it can lead to a mouldy odour.27 On the other hand
changes as a result of curing could occur during storage or shipment
of baled leaves from Asia to Europe or USA leading to higher quality
PEOs.27,48 The investigation by Gogröf produced results, which are
somewhat contradictory to those above.128 He found a 50% higher
oil content in leaves dried during 30 days relative to fresh leaves.
Possibly this is due to a “storage” effect (vide infra). Also deliberate
fermentation gave a slightly increased oil yield. Based on TLC, he
noted no qualitative changes as a result of drying or fermentation.128
The fact that changes are still occurring during storage was shown
in a study where leaves were stored for 210 days and analysed every
30 days for PEO content. Oil yield initially increased and reached a
maximum of 4.2% after 150 days of storage. Afterwards it declined
to 3.15% at 210 days, almost the same content as after 30 days of dry-
ing (3.05%).83,124 Fermentation was avoided. An explanation was not
given but slow evaporation could be the cause of the long‐term
decrease. In the same study shade drying and sun drying were com-
pared. The oil content was ~15% lower after sun drying. The effect
of storage on the oil content did not depend on the method of dry-
ing.83,124 According to Doraswamy sun drying is initially possible but
after a slight withering of the leaves, drying should be continued in
the shade.116 A storage of 150 days being optimal for the oil yield
was confirmed in an independent study.59 These authors suggested
that the increase up to 150 days is due to modest fermentation, which
weakens the cell walls and thereby facilitates oil recovery.59
Drying usually takes place by spreading the leaves out on a floor or
placing them in racks with proper ventilation and frequent turning to
avoid the growth of fungi.27,48,116 Doraswamy stated that leaves
should be dried to one fifth of their original weight to get a high yield
and superior quality.116 When comparing three different drying
methods, namely spread drying (8‐10 days), heap drying (10 days),
and spread (3‐4 days) followed by heap drying (2 days), the oil yields

VAN BEEK AND JOULAIN
were respectively 3.70, 3.35 and 3.87%.59 Ramya et al. reviewed sev-
eral drying studies and mentioned that oven drying at 30–50 °C during
half a day gives a similar oil yield as shade drying during 54 hr.55 This
was confirmed in another study where percentages of 2.46, 2.60 and
2.40% were reported for mechanical drying at 40 and 45 °C and shade
78
drying during 45 hrs respectively. Oyen cited a source stating that
drying at temperatures over 40 °C resulted in 80% loss of oil.48 In
another well‐documented study just the opposite result was found:
drying during 5 hr at 45 °C and 96 hr at 17–25 °C gave yields of
116
3.25 and 2.25% respectively. In both cases the final water content
was 17%. Masetto et al. compared drying periods of 0, 10, 14, 21
and 28 days and found the highest yield after 10 days of drying.112
Kongkathip compared oven‐dried leaves with leaves fermented
68
during 11, 34, 44 or 77 days. The percentages given for the oven‐
dried leaves (0.30%) and 34 days fermented leaves (1.04%) seem erro-
neous. The authors concluded that fermentation during 77 days gave
the highest oil yield (2.48%).68 Rulianah et al. have tried to degrade
cellulose and lignin in patchouli leaves by treatment with a white rot
fungus, Phanerochaete chrysosporium, in a liquid medium to increase
the oil yield.79 After a 7‐day treatment the oil content was 2.8% and
after 15 days 4.5%. The resinoid oil was isolated from the medium by
liquid‐liquid extraction (LLE) with hexane. A reference value obtained
by steam distillation of dried leaves was not given. Although the yield
is high, the procedure does not seem to be economically feasible on
a large scale. Another problem may reside in the olfactory properties
of the fermented oil. Yu et al. carried out a pretreatment of the leaves
with hemicellulase during 1 hr at 45 °C followed by steam distillation. It
gave a 41% higher oil yield than non‐treated leaves.129
Summarising, it is clear that proper drying prior to distillation
is absolutely necessary to obtain a good yield. However, what
exactly happens during the drying and possible fermentation and long‐
term storage in chemical terms and how this affects the odour quality
of the resulting PEO is still unclear after more than a century of research.
2.6 | Distillation Technique
Through a truly excellent microanalysis study, Henderson et al. showed
that in P. cablin volatile constituents are located in two different types
of cells.130 On the leaf epidermis one can find glandular trichomes.
Independent of its size, one leaf contains 60,000 trichomes and each
trichome contains 2 ng of essential oil. In the spongy mesophyll of
leaves one can find internal cells containing essential oil. There are
approximately 3 times more internal cells than external ones but as
the internal ones contain 3 times less essential oil, the amount of
PEO stored in both cell types is about the same. Qualitatively there
was no difference in composition. Stems and roots contained much
130
less essential oil than leaves. Another anatomic study postulated
that the internal mesophyll cells contain more sesquiterpenes although
no chemical analytical evidence was provided.131 These findings con-
127,132,133
firm much earlier observations as well as a recent micro-
scopic study.134 In 2013, two detailed electron microscopy studies
were performed in which up to six different types of glandular tri-
chomes, two forms of non‐glandular trichomes, and two types of inter-
135
nal glands were distinguished.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
9
As the oil in the internal structures is deeply buried and can hardly
pass the intact oil gland membranes, it is adamant that a drying step
precedes the hydro or steam distillation. More than a century ago de
Jong already remarked that otherwise only the oil in the outside glan-
dular hairs could be obtained.127 Quite remarkably, Gogröf reported
that fresh leaves yielded their oil faster than dried leaves. He came to
this conclusion by hydrodistillation experiments and microscopic stud-
ies of the leaves after distillation. Perhaps this contradictory result is
due to working with P. heyneanus instead of P. cablin. The exact identity
of the species used in his study is unclear.128 Recently the results of de
Jong were confirmed by hydrodistilling fresh, semi‐dry and dry leaves.
The distillation of fresh leaves was essentially over after 3 hours but
yielded only 1.82% of oil. For semi‐dry and dry leaves these figures
were 10 hours & 3.62% and 15 hours & 4.18% respectively.83,124
Bruns reported an almost 10% higher oil yield if the distillation time
was increased from 5.5 to 7.5 hours.75 But even after drying and grind-
ing, the high boiling constituents in PEO are difficult to distil, with
steam distillation being more effective than hydrodistillation.77 Von
Rechenberg reported a 10% higher yield with steam distillation starting
from the same leaves and also a different oil composition.77 Similarly
Doraswamy reported higher yields when steam distillation was used
versus hydrodistillation.116
The bottom line is that patchouli leaves require long distillation
times, a common feature for all sesquiterpene‐rich essential oils. De
Jong reported that it took several days before no more oil was
distilled and Gildemeister and Hoffmann mention 36 hr.26,127
Benveniste mentions a relatively short 6 hr for a steam distillation
to complete90 but Guenther cites steam distillation times of commer-
cial distilleries of 12–24 hr.27 He also remarked that higher steam
pressures reduce the distillation time and often but not always
increase the yield. A too high steam pressure or too long distillation
may affect the quality negatively. The steam pressure is best varied
during the distillation.27,55 By cohobation of the distillation waters,
the oil yield can be increased by 15%.27 Kumar showed that soaking
of dried leaves prior to hydrodistillation led to a 5% (after 12 hr of
soaking) to 10% (after 24 hr of soaking) higher yield of PEO.66
Recently Costa et al. reported that an 8 hr hydrodistillation of dried
leaves in a Clevenger apparatus statistically did not yield more oil
than a 1 hr distillation even though at first glance the percentages
of 4.8 (8 hr) and 3.3 (1 hr) suggest otherwise.108 As their conclusion
differs significantly from all others over the past century, it should be
treated with some reservation. Ramya et al. studied the effect of bed
packing densities of 8, 10 and 12 kg/m3 of the distillation still on the
oil content after 4‐6 hr of distillation time. The highest content of
2.78% was not surprisingly found for the lowest loading in combina-
tion with the longest distillation time although overall the differences
were small.55
2.7 | The presence of iron in patchouli essential oil
Iron ions can occur in PEO, as a result of the distillation process being
carried out in non‐stainless steel equipment or due to storage in iron
containers.27,136,137 The presence of iron ions causes the PEO to be
darker in colour,138,139 and can furthermore reduce the stability of
the oil.137,140 The presence of phenolic or enolic constituents or a high
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
10 VAN BEEK AND JOULAIN

acidity increases the iron content27,138 so we hypothesise that the iron 1

H‐NMR.152,153 Already in 1967 Tsubaki applied capillary GC to better
content in a PEO might be partially related to its pogostone content. separate PEO hydrocarbons.154
Harunsyah reported on a PEO having an iron content of 384 Over the next 15 years several fragrance companies, like IFF,155
137,140 141
ppm, while 25 ppm is the specified maximum. Some compa- Roure Bertrand Dupont,156 and Henkel KGaA,75 carried out in‐depth
nies offer PEOs containing iron and iron‐free qualities.142 Iron ions analyses of patchouli oil. Most of our current knowledge on its constit-
91
can be removed by treatment with citric acid, tartaric acid or EDTA, uents still stems from these investigations. The main motivation was to
oxalate,143 by molecular distillation,27,142,144 or by filtration over ben- find out which molecules are responsible for the unique patchouli
tonite.145 Holdsworth‐Haines described the use of tartaric acid to odour (see also § 5.1 on odour) and GC‐olfactometry (“sniffing port”
remove either copper or iron from PEO.138 Copper‐containing PEOs analyses) was applied several times to PEO. In this period GC com-
138
are greenish in colour. bined with mass spectrometry became commercially available and pro-
vided hitherto unknown analytical capabilities in the essential oil field.
13
C‐NMR was introduced during this period as a new and very useful

3 | ANALYTICAL INVESTIGATIONS OF spectroscopic technique.

P A T CH O U L I E S S EN T I A L O I L
3.1 | History of patchouli oil analysis and used
analytical techniques
Several periods regarding patchouli oil analysis can be distinguished.
The first period runs from 1840–1960 during which analyses were
mostly focused on characterisation of the oil by parameters like spe-
cific gravity, refractive index,146,147 optical rotation, colour, odour
and solubility (Table 2). Separation of compounds proceeded by vac-
uum distillation and crystallisation, and structure elucidation took place
by highly elegant but nonetheless tedious and time‐consuming chemi-
cal degradation and derivatisation studies. The purity of products was
most often assessed by boiling points, elemental analysis and mixed
melting points (m.p.) with reference substances. Although patchoulol
had already been isolated in the 19th century, not a single structure
of the major sesquiterpenic constituents was known prior to
1960.26,27 However systematic investigations were on‐going.148-150
and in the period 1950–1960 column chromatography as well as UV
The fourth and last period runs from approximately 1985 till our
present time. GC‐MS is now widely available and mass spectral data-
bases (Wiley, NIST) come with the purchase of the instrument. This
resulted in quantitatively many analyses, of which many are qualita-
tively of lower quality than before. High‐speed GC on 10 m × 0.1 mm
× 0.2 μm columns has been applied on PEO.157,158 In spite of run times
shorter than 5 min with fair resolution, in practice this approach never
became popular. Occasionally new compounds in patchouli oil are
reported159,161 or the structure of a known compound corrected.162
Vibrational circular dichroism (VCD), which allows the distinction of
enantiomers even when they lack a UV chromophore, was applied in
the latter study. As the VCD spectrum can be theoretically calculated,
it allows the determination of the absolute configuration. Nowadays
structural proof is provided by 2‐dimensional NMR rather than by total
synthesis. Chromatographic resolution has potentially dramatically
increased by the commercial availability of GC×GC‐TOFMS (compre-
hensive two‐dimensional gas chromatography with time‐of‐flight mass
spectrometry for detection). Fragrance companies continue to be inter-
ested in patchouli oil and patchouli odour.162-165

TABLE 2 Physicochemical properties of patchouli essential oil

166 167 141 27
Source
Specific gravity (g/mL) 0.954–0.983 0.952–0.975 0.950–0.975 0.940–0.995
Refractive Index 1.4990–1.5120 1.5050–1.5150 1.507–1.515 1.505–1.5160
Optical rotation −43°–−66° −40°–−60° −48°–−65° −40°–−72°
Miscibility with 90% ethanol mixture of 10:1 90% ethanol: mixture of 10:1 90% ethanol:oil gives 1:2 to 10:1 90% ethanol:
oil is clear a diaphanous solution or light oil is clear
opalescence
Colour yellow to reddish brown light yellow to reddish brown

and IR spectroscopy were used for the first time.149 Gogröf was the According to Guenther higher quality oils exhibit higher absolute
first to apply a chromatographic method in an analytical fashion to specific gravity, refractive index and optical rotation values and good
PEO (ascending thick layer chromatography with silica gel solubility.27 A high specific gravity and optical rotation together indi-
"chromatostrips", an early version of TLC).128 cate a high patchoulol content and consequently a lower hydrocarbon
The period from 1960–1970 saw the identification of several content, which will provide a more pleasant aroma. In the 1930s, so
major constituents like patchoulol, α‐guaiene, α and β‐patchoulene, called “Singapore oils” were in high demand and showed a higher spe-
seychellene, and α‐bulnesene. Synthesis played an important role in cific gravity and better miscibility than “Java patchouli oil”.168 In
these investigations and an X‐ray study finally solved the structure of another case, a PEO from the Seychelles was rejected on the basis of
patchoulol.151 Newly introduced techniques included both analytical wrong figures for specific gravity, refractive index and optical rotation
and preparative packed column gas chromatography, and in spite of a fair odour and good solubility.169

VAN BEEK AND JOULAIN
Nowadays PEO as well as the leaves of P. cablin occur as mono-
graphs in the Chinese Pharmacopoeia.170,171 Patchouli leaves are iden-
tified by microscopy and TLC of the contained essential oil with
patchoulol as reference substance. For patchoulol a minimum content
of 0.10% based on dry leaves is specified.171 PEO is identified by its
colour, smell, specific gravity (0.950 ‐ 0.980) specific rotation
(−43°–−66°), refractive index (1.503‐1.513), TLC on silica with
patchoulol and pogostone as reference substances and the absence
of materials insoluble in 90% ethanol (1:10 ratio). A minimum content
170
of 26% is specified for patchoulol.
3.2 | Considerations for the evaluation of published
patchouli oil analyses
All papers describing the structure elucidation of individual compounds
or chemical analyses of the total essential oil of Pogostemon cablin
(patchouli oil) were carefully checked with two aims in mind. Aim 1
was to arrive at a listing of all compounds, which – with a greater
(Table 3A) or lesser degree (Table 3B) of confidence – occur in PEO.
Aim 2 was to determine the qualitative and quantitative variation of
the major constituents (Table 4). Papers devoted to solvent or super-
critical extracts were excluded as they do not deal with essential oils
and cannot be compared quantitatively with essential oils by GC, since
extracted non‐volatile compounds do not elute. By definition essential
oils are obtained via steam distillation (mostly industrial scale) or
172
hydrodistillation (lab scale, e.g., Clevenger apparatus). A second
consideration was that there is currently no significant market for
odorant‐rich patchouli extracts. Finally, the benzene extract, which
173,174
was mostly used in the past, is now obsolete.
Although essential oil analysis appears simple and straightforward
as it only involves the injection of a diluted solution of the essential oil
into a GC‐MS, in our opinion it is not. This is especially true for oils
consisting mostly of sesquiterpene hydrocarbons and oxygenated ses-
quiterpenes, like for instance PEO. First of all, the chromatography can
be poor, e.g., due to severe overloading of the column or too short
retention times (low k' values), and several examples of such shortcom-
ings in the patchouli literature were encountered. This will automati-
cally result in more co‐elution and additionally the mass spectrum
can be affected. Less credit was given to such analyses. The paper by
Frister is an example of too short retention times.175 Additionally there
are wrong mass spectral assignments and errors in the legend.175 The
real pitfall in essential oil analysis however, lies in the data processing.
Commercial mass spectral databases, such as NIST and Wiley, are
not dedicated to essential oils, which means that many sesquiterpene
spectra do not occur in these databases but spectra of many more
non‐volatile constituents do occur. As the library search algorithm will
always generate some fit, albeit at 30% similarity, this can lead to the
supposed identification of rather odd compounds in patchouli oil such
as farnesyl pyrophosphate (ionic, non‐volatile), thyroxine (a non‐
volatile human hormone), 5‐azulenemethanol (a blue compound;
patchouli oil is never blue), tetrahydrosmilagenin (a non‐volatile
steroid‐triol), squalene (non‐steam distillable, human skin
constituent), 1,4,7,10,13,16‐hexaoxacyclooctadecane (a crown ether),
3‐hexadecyne (so far not known as a natural product), dibutyl phthal-
ate (plasticizer), anthracene (polyaromatic hydrocarbon), 4,5‐
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
11
dimethoxy‐2‐methylphenol (not less than 34.6% of the oil), trilaurin
(39 carbons, non‐steam distillable), glycyl‐L‐proline, 2‐hydroxy‐5,6‐
epoxy‐15‐methyl‐pregnan‐20‐one or butylated hydroxytoluene
(synthetic antioxidant) to name a few. Papers reporting such com-
pounds were given less credit with regard to other identifications.
As mass spectra of structurally different sesquiterpenes can be
near identical, the opposite is also true: even a fit of 99% does not
always guarantee that a compound has been correctly identified. We
came across many examples of misidentifications of this kind. Although
in retrospect one can of course never prove that a certain constituent
did not occur in a particular oil, based on the over 100 patchouli
analyses studied, we have excluded isolated reports of – sometimes
major – compounds not mentioned by other investigators. An example
is germacrene A reported at a concentration of 11.73% while for the
almost co‐eluting α‐bulnesene a concentration of only 0.86% was
given in the same paper. α‐Bulnesene is one of the four major constit-
uents of PEO. Superficially the two mass spectra are similar. In 2017,
an “improved” patchouli plant containing 40.41% β‐selinene as major
constituent was reported. Based on the depicted chromatograms, most
likely an identification error was made.
An additional problem of complex oils, like patchouli, is that inev-
itably some compounds (partially) co‐elute, which changes the mass
spectrum of the major contributor and obscures the spectrum of the
minor one, resulting in turn in a wrong assignment. By careful
deconvolution this can be corrected to some extent but this requires
mass spectral expertise, which sometimes appears to be absent. Also,
long listings of identified trace constituents were considered as some-
what suspicious for the reasons given above. In our view, still more is
not known about the trace constituents (concentration from 0.01‐
0.5%) of PEO than what is known. Occasionally authors have reported
on the occurrence of a specific enantiomer, e.g., in 2016 both (−)‐
caryophyllene oxide and caryophyllene oxide (later eluting) were
reported in one and the same analysis of patchouli oil. However, in
all such cases a non‐chiral column was used making such assignments
impossible and thus the report less credible. The same holds true for
the same compounds reported twice, or even thrice, in a single analysis
and the inclusion of vague compound names such as “trans‐tricyclo” or
“isobutyrate” or “apha‐guanine”.
Similarly, incidental reports of sesquiterpenes belonging to biosyn-
thetically very different classes from those occurring in PEO were
disregarded until more convincing evidence than just a mass
spectrum is provided. The major sesquiterpenes occurring in
patchouli oil belong to the guaiane, α/γ‐patchoulane, β/δ‐
patchoulane, deoxypatchoulol, seychellane, and caryophyllane classes.
Minor constituents belong to the elemane, eudesmane, germacrane,
humulane, farnesane, cycloseychellane, nortetracyclopatchoulol and
copaane classes. Although we cannot exclude the occurrence of other
classes, until further proof is provided we treat the identifications of,
for example, (−)‐eromophilene, cis‐thujopsene, trans‐longipinocarveol,
longifolenaldehyde, (+)‐α‐longipinene, aristolone, 4,5‐diepiari-
stolochene, β‐ylangene, valencene, nootkatene, dihydrokaranone, γ‐
muurolene, γ and δ‐cadinene, 4‐oxo‐14‐norvitrane, β‐chamigrene,
and α‐bergamotene on biosynthetic grounds as doubtful.
Retention indices (RIs or LRIs = linear retention indices) are neces-
sary to identify sesquiterpenes with a much greater degree of

12
confidence than by mass spectra alone. Tables with RIs of sesquiter-
penes are available.176-178 However we noted several papers where
RIs seem to have not been experimentally determined but rather sim-
ply copy/pasted from the aforementioned databases. Such papers
have been treated cautiously. Also, papers reporting impossible RIs
such as RI = 932 for patchoulol or RI = 1939 for l‐(+)‐ascorbic acid
2,6‐dihexadecanoate (anyhow not a steam distillable compound) hav-
ing 37 carbons were considered to be less reliable. Papers reporting
oxygenated sesquiterpenes eluting before sesquiterpene hydrocar-
bons, or tridecane eluting 7 min after tetradecane (in a 2016 paper)
have been treated with some reservation. RIs can somewhat vary even
if recorded on the same stationary phase depending on carrier gas,
inlet pressure, column length and diameter, film thickness, analyte con-
centration, detector (FID versus MS) and especially the temperature
gradient.179 Peak reversals are possible due to using different temper-
ature gradients or by going from gradient to isothermal elution. Polar
phases (such as PEG 20M), which are often preferred when analysing
essential oils, are prone to produce more variable and thus less reliable
RIs upon ageing and depending on manufacturers. Thus, like mass
spectra, RIs should be used wisely.
Errors in identifications, reported mass spectra and RIs do occur in
the literature and through error multiplication ("domino effect") can
further complicate essential oil analyses. The most notorious error of
this kind relates to the identification of seychellene as γ‐patchoulene
by Wenninger et al. in 1966.153 Presumably based on this persisting
error, Hussa et al. still misidentified in 2011, so 45 years (!) later,
seychellene as γ‐patchoulene. This particular confusion in the litera-
ture is further aggravated by an unknown component, having the
same mass spectrum as γ‐patchoulene, almost co‐eluting with δ‐
patchoulene (see also § 3.3 on identification). In spite of claims by
some authors, reference standards are regrettably not available for
most sesquiterpenes occurring in PEO.
Still another type of error that can occur in patchouli analysis is
either a wrong botanical identification or a mix‐up of samples in the
lab. For instance, in a 2016 paper, carvacrol (47.5%) and p‐cymene
(15.2%) were reported as the main constituents of patchouli oil. Not
surprisingly, neither patchoulol nor any of the other constituents typi-
cal of patchouli oil were reported. This shows a total lack of any liter-
ature research by the authors and this paper has not been cited in
this review but does occur in the SI. Possibly a botanical misidentifica-
tion also played a role in the atypical oils reported on by Sugimura et al.
in the 90s.74,180 Since then no one else has reported near 20% of
germacrene B and longifolene in PEO.
A further problem is the occurrence of extraneous constituents in
patchouli oil, by inadvertent contamination of leaves of P. cablin with
other materials like weeds prior to steam distillation, by a lack of
cleaning of the distillation equipment between charges or by wilful
adulteration before or after distillation. An example of the latter is
the addition of gurjun balsam, which contains compounds belonging
to the aromadendrane class (see § 4 on adulteration). Several authors
have reported the occurrence of acetophenone in patchouli oil.
Acetophenone is the main constituent (51%)181 of P. heyneanus and
48
this species is used to adulterate P. cablin. Only small amounts of P.
heyneanus will be needed to be able to detect acetophenone as a minor
constituent in genuine PEO. Murugan et al. in their comparative study
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
of both Pogostemon species were unable to detect acetophenone in P.
cablin oil.181 Probably several unlikely compounds detected in PEO
have been correctly identified and originated from a contamination.
This however was in most cases impossible to prove.
Finally we encountered one odd case where analytical data of one
set of authors182 were copied five years later by a pair of unrelated
authors without providing any remark in the experimental or a citation.
Furthermore, the same pair of authors (Kusuma and Mahfud) subse-
quently republished these data in 2017 with unexplained changes
and without citing their own paper from 2016. Together these facts
made us entirely disregard the six very similar papers by these two
authors, however these references like all others rejected can be found
in the SI.
3.3 | Constituents of Patchouli Essential Oil
Based on all criterions mentioned above, we weighed the quality of
each report and gave more value to – in our opinion – high quality
reports and completely disregarded (and neither used nor cited) lower
quality analyses. However, all references relating to non‐used patch-
ouli analyses can be found in the supplementary information. In retro-
spect, it was possible to correct several clearly wrong assignments,
especially when a chromatogram was available. In such cases,
corrected data have been used. We have also compared the combined
results qualitatively with independent GC‐MS analyses of genuine
PEOs carried out in the laboratories (Robertet and Wageningen Uni-
versity) of both authors using dedicated essential oil mass spectral
databases.
The outcome of the excellent, good and fair reports has been used
to compile a listing of compounds, which occur in PEO with a high
degree of certainty (Table 3A) or with a reasonable degree of certainty
(Table 3B). This information is qualitative, i.e., whether a constituent
occurred at a high or low concentration did not play a role. So actually,
Tables 3A and 3B contain several compounds, which cannot be
detected in total PEO, not even by an expert with a modern GC‐MS.
This is especially true for carboxylic acids, phenols and amines
(alkaloids), which can be easily separated and significantly enriched by
LLE. In the literature, hundreds of compounds have been reported to
occur in patchouli oil. If a constituent has been reported in an – in our
view – dubious paper or has been reported only once or twice, the con-
stituent did not make into Table 3B. First, additional independent proof
should be provided. Some compounds or groups of compounds merit
more attention from an identification point of view, i.e., because the
identification is troublesome or because of a structural revision. These
are discussed in the following paragraphs after which Table 3A and 3B
with 72 positively and 58 tentatively identified patchouli constituents
respectively follow. Identifications reported by scientists from the
fragrance industry deserve special attention,155,165,183-185 since they
have not been published in peer‐reviewed journals on the basis of
detailed spectroscopic data recorded from isolated pure substances.
As this missing information may exist as proprietary information of
these companies, many of these identifications could be correct.
However, as they do not satisfy the requirements of this journal, we
have therefore included them in Table 3B.

VAN BEEK AND JOULAIN
3.3.1 | SQHCs in general
Sesquiterpene hydrocarbons (SQHCs) constitute a large fraction of
PEO. Hefendehl and Welch et al. both mention 62%,186,187 while
Ohloff et al. report 40‐60%.52 Our in‐house analyses of more than 20
commercial samples show a wide range of variation (49‐73%, mean
163
59%). The literature data in Table 4 based on 100+ PEO analyses
yield an average of 47% excluding γ and δ‐patchoulene and unidentified
SQHCs so approximately half of the oil. In extreme cases the SQHC con-
tent can be > 80% or < 25%. As discussed in § 3.2, the identification of
sesquiterpenes, including SQHCs, is wrought with difficulties leading to
many misidentifications in PEO. Ideally identification should be based on
an identical EI‐MS according to both the computer algorithm and a
skilled human observer and identical retention times to an authentic ref-
erence substance on two different GC phases. Thus, as only few SQHCs
are commercially available, identifications are rarely ideal.
In the very first chemical study of PEO in 1869, it was reported that
patchoulol 33 could be easily converted to a SQHC, which must have
been a mixture of patchoulenes.188 The name “patchouline” was coined
for this product in 1877,189 which was later changed to “patchoulene”.
The first attempt to isolated pure SQHCs by repeated fractional distil-
lation from PEO took place in 1904.190 Based on the reported boiling
points, fractions 1 and 2 might have been (partially pure) α‐patchoulene
24 and α‐bulnesene 15 respectively. After partially successful studies
on probably again α‐patchoulene and α‐bulnesene in 1950 by Šorm
et al.,149 the identification of the main SQHCs in PEO started in earnest
in 1962. 152,191
The identification of α‐bulnesene 15, α‐guaiene 16 and
β‐patchoulene 21 was based on a comparison by GC, UV, IR and NMR
with isolated dehydration products of guaiol and bulnesol. The
contemporary study by Büchi et al. on the formation of the various
patchoulenes from patchoulol 33 must have been helpful. 192
A few
years later, Tsubaki et al. used a combination of the new capillary GC
technique, column chromatography (including AgNO3 complexation),
preparative GC, NMR and IR to confirm the presence of α‐bulnesene
15, α‐guaiene 16 and β‐patchoulene 21, and establish that of
α‐patchoulene 24.154 They also identified β‐elemene 7 and β‐
caryophyllene 18, the latter being a genuine PEO constituent as con-
firmed by others.153,193,194 Additionally Tsubaki et al. detected but
not identified a tricyclic sesquiterpene “hydrocarbon G” having a termi-
nal methylene according to its IR and NMR spectra. The mass spectrum
showed the base peak at m/z 122.154 It was proven to be identical to
seychellene 26, which was simultaneously isolated and structurally
195,196
identified by Wolff et al. In his study of a PEO produced
in Brazil, Peyron reported in addition to most of the SQHCs
mentioned above, α‐elemene and λ‐patchoulene. In retrospect, the
latter two compounds were most likely β‐elemene 7 and a mixture of
seychellene 26 and δ‐patchoulene 22, respectively. 194
Teisseire
reported in 1973 the isolation of 13 sesquiterpene hydrocarbons, of
which 8 were identified. Of those, only α‐humulene 5 had not been
reported before in PEO.156,193 His unidentified “hydrocarbon B” was
later shown to be δ‐patchoulene 22. 197
A few minor SQHCs in PEO
merit special attention and are discussed individually at the end of this
section.
Faraldos et al. have commented on the confusing nomenclature
198
of the patchoulane sesquiterpenes. As the skeletons of
α/γ‐patchoulane (tricyclo[5.3.1.01,5]undecane), β/δ‐patchoulane
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
13
(tricyclo[6.2.1.02,6]undecane) and patchoulol (tricyclo[5.3.1.05,10]
undecane) all share the same cyclodecane ring (C1 to C10) and differ
only in the bridging positions of the isopropyl group, i.e., C11 with its
two geminal methyl groups, and the zero carbon connections (second-
ary bridge), they proposed shorthand names to name the three
patchoulane skeletons unambiguously.198 Based on the location of
the C11 bridge and the second ring fusion, they distinguish
[1,7:5,10]‐, [1,7:1,5] and [7,10:1,5]patchoulanes. Patchoulol 33 would
thus be named [1,7:5,10]patchoulan‐1‐ol.198
The chemical structures of most of the major SQHCs have been
unambiguously confirmed by either total synthesis, like for seychellene
and cycloseychellene,187,199 or hemi‐synthesis, e.g., α‐guaiene 16 and
aciphyllene 17 from guaiol.200 Nowadays, NMR is often replacing full
or hemi‐synthesis for proof of structure but errors are possible and
pogostol 37 is a telling example (vide infra). High resolution NMR
and MS data have been published for many isolated SQHCs, e.g., for
α‐guaiene, α‐bulnesene, seychellene and α‐patchoulene, albeit in a
rather confusing article requiring errata.201,202
Some of the PEO SQHCs, e.g., β‐patchoulene 21 and α‐
patchoulene 24 are suspected to be artifacts formed by dehydration
of patchoulol 33 and subsequent Wagner‐Meerwein rearrangements
during steam distillation.52 Invariably when patchoulol is dehydrated,
depending on the conditions various mixtures of patchoulenes and
other rearranged hydrocarbons result. For instance, when treated with
POCl3 in pyridine, a mixture of γ‐patchoulene 23 and α‐patchoulene 24
resulted in 91% yield. When the mixture of 23 & 24 or patchoulol 33
was treated with concentrated sulphuric acid in ether, β‐patchoulene
21 was formed.192 And when heated in refluxing hexane in the
presence of acidic Amberlite‐IR‐120H, patchoulol is transformed
in 77% yield into a mixture containing 55% β‐patchoulene 21 and
5% δ‐patchoulene 22, among other unidentified hydrocarbons.203
Interestingly, small amounts of seychellene 26 are also formed under
these conditions, together with ε‐patchoulene 76.163 The latter has
been tentatively identified as a very minor constituent in PEO and might
well be an artifact formed by isomerisation of β‐patchoulene and/or
δ‐patchoulene during distillation.163 Although the above conditions
differ from those during steam distillation, almost certainly, depending
on pH and temperature, dehydration of patchoulol followed by
carbocation rearrangements take place during steam distillation.
However, what happens exactly during the fabrication of PEO and the
influence of steam distillation on its qualitative and quantitative compo-
sition has never been investigated in detail with modern techniques.
Aromadendrene and cadinane SQHCs have been frequently
reported in PEO but we consider these reports erroneous (see also
§ 3.2). An example is the report by Ishihara et al. (PEO SQHC fraction
containing 4% gurjunene and 1% δ‐cadinene).204 The presence of
the former class is the result of adulteration.205,206 The mentioning
of δ‐cadinene in concentrations of 2‐3%40 in four PEOs is probably
due to a misidentification of aciphyllene 17 (Lawrence, personal
communication).
3.3.2 | Aciphyllene
Aciphyllene (guai‐4,11‐diene) 17 is always present in a few percent in
patchouli oils. It elutes just before the major constituent α‐bulnesene

14
15 on non‐polar columns. It was independently identified by Saritas
and by Joulain,163,207 and later confirmed by others.208 In the structure
given by the former two authors the 2‐propylidene side chain at C‐7
has the α‐configuration, in contrast to almost all other sesquiterpenes
in PEO. When aciphyllene was first synthesized by Blay et al., its spec-
209
tral data did not match those of the natural product. Subsequently
its structure was revised to 17 (Table 3A), i.e., with the C‐7 substituent
in the β‐position. The structural revision was confirmed by Srikrishna
and Pardeshi,210 and later by synthesis.200
3.3.3 | α‐Copaene
This hydrocarbon has been cited as a possible marker for contamination
or adulteration. Indeed, the earlier mentioned ISO norm states that the
content of this hydrocarbon should not exceed 1%, which suggests that
higher levels would indicate some adulteration, possibly with copaiba
balsam (Copaifera spp.). While authentic PEOs have been found to
contain 0.1% α‐copaene 25,123 the authors have recorded lower
percentages or even undetectable levels of this SQHC in many oils
distilled and analysed by us and PEOs with α‐copaene levels >0.5%
should indeed be treated as suspicious in terms of possible adulteration.
3.3.4 | Cycloseychellene
211
The rare tetracyclic cycloseychellene 27 was first reported in 1973.
One year later Dhekne and Paknikar showed that Tsubaki's “hydrocar-
154 212
bon E” (Figure 1) was actually cycloseychellene. Its structure was
revised in 1981 on the basis of high field NMR studies,213 and total
199
synthesis.
211
FIGURE 1 Left: initial (wrong) structure of cycloseychellene.
Right: Revised and correct structure of cycloseychellene 27213
3.3.5 | γ‐Patchoulene
The IR spectrum initially assigned to γ‐patchoulene 23 isolated from
PEO actually corresponded to seychellene 26.214,215 The IR spectrum
of authentic γ‐patchoulene obtained by dehydrating patchoulol with
POCl3 and preparative GC (pGC), was reported later.215 This spectrum
is identical to that of γ‐patchoulene isolated from PEO in July 2000 by
one of the authors.163 Indeed the IR spectra of γ‐patchoulene 23 and
seychellene 26 show striking similarities, whereas these hydrocarbons
are easily distinguished by their mass spectra and linear retention
indices (LRIs). Although frequently reported as a constituent of PEO,
whether γ‐patchoulene genuinely occurs in patchouli leaves or is
merely an artifact, remains to be determined. Tsubaki et al. could not
detect it in PEO but they could detect it in distillation fractions of
PEO. According to them it elutes just after α‐bulnesene on a HB‐
2000 stationary phase.154 Teisseire confirmed that γ‐patchoulene
does not occur in PEO and that its retention time in GC is totally differ-
193
ent from seychellene. On a dimethylpolysiloxane phase (syn. DB1,
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
SE30 or CP Sil5) an LRI of 1497 is reported, i.e., close to aciphyllene
(LRI=1495) and α‐bulnesene (LRI=1503).177 Unfortunately often
“γ‐patchoulene” is reported to elute close to α‐patchoulene and
δ‐patchoulene, probably due to the elution there of a still unknown
SQHC having almost the same mass spectrum as 23.
3.3.6 | η‐Selinene
η‐Selinene 75 (ent‐isomer of cascarilladiene) has been tentatively
identified as a very minor SQHC in PEO (Joulain, unpublished). It is
easily overlooked as it coelutes with cycloseychellene and/or
β‐caryophyllene on non‐polar phases. However, on a polar Wax phase,
it can be observed close to cycloseychellene and α‐guaiene.
3.3.7 | Oxygenated sesquiterpenes in patchouli
essential oil
Patchoulol
Patchoulol (syn. patchouli alcohol) was first isolated in crystalline form
by Gal in 1869 as the major component of PEO.188 The first correct
elemental composition (C15H26O; adjusted from the C30H26O2 given
in the paper) of what was then named "patchouli camphor" was
derived 8 years later.189 In retrospect amazingly correct, the com-
pound was suspected to be the hydrated hydrocarbon (C5H8)3, i.e.,
C15H24 + H2O. Wallach and Tuttle confirmed these data in 1894.216
They also suggested that patchoulol is a tertiary alcohol, a proposal
which found support 10 years later.217 Next Semmler and Mayer con-
cluded that patchoulol is a tricyclic alcohol on the basis of its molecular
refractivity.218 The first – incorrect – structure A (Figure 2) for
patchoulol was published by Treibs.148 Unaware of carbocation rear-
rangements, he concluded from the fact that dehydration produced
some (blue) guiazulene that patchoulol must possess a bridged 5‐7 ring
system. This work was continued by Büchi et al. with a series of
chemical degradations of β‐patchoulene 21, γ‐patchoulene 23 and
α‐patchoulene 24.192 The isolation and the structural determination
of 21, 23 and 24 obtained by pyrolysis of the acetate and the forma-
tion of β‐patchoulene 21 by dehydration of patchouli alcohol
with H2SO4, I2, or H3BO3, led to the – incorrect – conclusion that
patchoulol possessed the tricyclic structure B (Figure 2). With struc-
ture B in mind, an elegant partial synthesis of patchoulol was designed
from (+)‐camphor, via an epoxide of 24, which was expected to lead to
optically active "patchouli alcohol" B but in reality led to the true
natural product 33!219 However shortly thereafter, structure 33 was
finally and unambiguously assigned to patchoulol on the basis of an
X‐ray crystal analysis151 of the known bright red chromic acid ester.220
The apparently contradictory results of both the chemical elucidation
of the structure of patchoulol and of its partial synthesis from (+)‐
camphor were explained in a subsequent article.221 The unexpected
reverse skeletal rearrangement of an epoxide, which was overlooked
by the authors, caused faith to be placed in the wrong structure B
instead of 33. This historical case has been attractively reviewed by
renowned organic chemists.222,223 Faraldos et al. studied the
biosynthesis of patchoulol in considerable detail through the use of
deuterium labelled farnesyl diphosphate.198 As part of these studies,
they published the most extensive NMR study of 33.198

VAN BEEK AND JOULAIN
While patchoulol is not specific for Pogostemon species, its occur-
rence in high concentrations is nevertheless scarce in other essential
oils, and mostly restricted to certain Valeriana species,224 for example
26.2% in Valeriana celtica leaf oil,225 up to 50% in Valeriana wallichii
rhizome oil,226 in Valeriana pancicii root and rhizome oil,277 and up to
66.7% in Valeriana jatamansi rhizome oil.228 Adulteration of V. celtica
and V. jatamansi with PEO supposedly occurs, however as in the above
cited studies genuine plant material was used, there is no doubt that
patchoulol is a true constituent of several Valeriana essential oils.
Nowadays, purified patchoulol obtained by fractional distillation of
PEO and subsequent crystallisation is commercially available in multi‐
kg quantities for fragrance applications under the trade name
Healingwood.229
FIGURE 2 Left: first (wrong) structure A of patchoulol.148
Middle: second (wrong) structure B of patchoulol.192,219
Right: correct structure 33 of patchoulol151
3.3.8 | Norpatchoulenol
Norpatchoulenol 34 was first reported in PEO by Teisseire's group in
1971.230,231 Its structure and absolute configuration were determined
by a correlation with cyperene232 (Scheme 1) and an X‐ray analysis of a
233
crystalline bromo‐ketone derivative. Later Mookherjee claimed that
norpatchoulenol had already been known to his employer since
1966.155 The concentration of 34 in PEO is on average 1% but the
actual percentages vary from 0.11 to 4% (see Table 4).
SCHEME 1 Possible biosynthetic route to norpatchoulenol 34 starting from patchoulol 33 via intermediates C and D according to Ourisson.234
Far right: structure of cyperene
A biogenesis of 34 from patchoulol 33 (Scheme 1) has been pro-
posed by Ourisson, based on forcible gastric feeding of 33 to rabbits
and dogs, generating diol C and hydroxy‐acid D as hydrolysable glucu-
ronides present in the urine of the animals.234 Diol C was also pro-
235
duced by microbial hydroxylation (bacteria or fungi). Catalytic or
chemical oxidation of diol C furnished hydroxy‐acid D and the latter
was converted to 34 by lead tetraacetate in pyridine in the presence
of cupric acetate. However, neither C nor D has been identified so
far as putative precursors of patchoulol in PEO or any patchouli
extract. Also, our analyses of many PEOs, do not point in the direction
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
15
of a relation between patchoulol and norpatchoulenol concentrations.
On the other hand, the concentrations of norpatchoulenol and
nortetracyclopatchoulol 35 do seem to be related suggesting a com-
mon C14 precursor. Thus, the biosynthesis of norpatchoulenol 34
(as well as 35) in patchouli leaves remains unknown to date.
3.3.9 | Nortetracyclopatchoulol
The structure of nortetracyclopatchoulol was originally published as a
trace constituent (0.001%) with a typical patchouli odour, which was
even slightly stronger than that of patchoulol and norpatchoulenol.155
As an X‐ray study failed, in addition to the obtained GC, IR, MS and
NMR data, the authors performed a correlation with norpatchoulenol.
Treatment with a strongly acidic ion exchanger gave indeed the
expected norpatchoulenol after which the authors concluded that E
(Figure 3) represented the trace constituent. The configuration of the
cyclopropane ring was left undecided. For the next 33 years this was
assumed to be the correct structure. However, in 2014, Zaizen et al.
published structure 35, also a nortetracyclopatchoulol, as a novel
odour‐active compound in patchouli oil. They performed a rigorous
13
NMR study including INADEQUATE C‐13C measurements, which
technique was not available in the early 80s.162 Sniffing port gas chro-
matography was used to pinpoint this minor constituent. However, as
no nortetracyclopatchoulol was detected in their study and the mass
spectrum for 35 was identical to that of nortetracyclopatchoulol, we
reason that Zaizen et al. in fact reisolated nortetracyclopatchoulol.
Additional proof for this hypothesis can be found in the fact that in
our own PEO analyses, only one peak with the same mass spectrum as
35 can be observed and on a WAX phase this compound elutes just
after patchoulol and well before pogostol (Joulain, unpublished), i.e.,
at the same position as in the Wax chromatogram of Zaizen et al. This
would imply that structure E is wrong. This is certainly possible as it
would be very difficult to distinguish between E and 35 with low res-
olution NMR without having both compounds available. It is further
likely that 35 would be converted to norpatchoulenol upon treatment
with acid. Thus, the important odour‐active nortetracyclopatchoulol
has structure 35 and occurs as such in Table 3A. The absolute
configuration was proven by vibrational circular dichroism (VCD).162
As can be seen in the chromatogram of PEO B (Figure 7b),
nortetracyclopatchoulol (peak marked ntP at 44.5 min) is not necessar-
ily a 0.001% trace constituent but can occur at concentrations of 0.5%
or higher. Therefore, in some oils it might contribute significantly to
the overall patchouli odour.

16
155
FIGURE 3 Left: initial (wrong) structure E of nortetracyclopatchoulol.
Right: Revised correct structure of nortetracyclopatchoulol 35162
3.3.10 | Pogostol
Although pogostol has been reported to occur in more than 30 essen-
tial oils including PEO, its identification has been rarely supported by
more evidence than mere GC‐MS data. However, PEO is the exception
to the rule as several authors have reported also NMR data and its
occurrence in PEO is beyond doubt. Pogostol was first isolated by Pfau
236
and Plattner in 1936. They converted it to the blue guiazulene by
the dehydration/dehydrogenation method, which they had just
invented. In 1968 Hikino et al. proposed structure F (Figure 4), i.e.,
without any stereochemistry.237 The stereochemistry at C4, C5 and
C7 was partially clarified (structure G, Figure 4) by Teisseire et al. on
the basis of convincing chemical correlations with both bulnesol and
α‐bulnesene whose absolute stereochemistries were already known
at that time.156,193 Unfortunately he did not provide any specific
optical rotations in his Experimental. Ten years later Itokawa et al.
isolated a sesquiterpene alcohol from Alpinia japonica rhizomes and
identified it as pogostol on the basis of having identical IR and low field
1
H NMR data as an authentic reference.238 Although they overlooked
Teisseire's findings and did not show any stereochemistry, they did
13
report C NMR data, which proved to be very useful in subsequent
investigations by other authors. In 1987 without further explanation,
Teisseire reported structure H (Figure 4) for pogostol.239 In 1997,
Fleischer et al. reported pogostol O‐methyl ether from Artabotrys
stenopetalus with the same stereochemistry as in H.240 They elucidated
the structure on the basis of extensive NMR studies including HMBC
and NOESY spectra. In 2000, during the course of an in‐depth
analysis of vetiver essential oil, Weyerstahl et al. isolated and
13
identified pogostol by comparing their C‐NMR data with those of
FIGURE 4 Various postulated, partially inaccurate, structures of pogostol 37
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
Hitokawa.241 By NOED NMR experiments (not described in the
Experimental) they arrived at the same structure H for pogostol as
published earlier by Teisseire. The same conclusion was reached by
Stierle et al.242
In the same year the structures for pogostol and its methyl ether
were challenged by Booker‐Milburn et al. after their total synthe-
sis.243 On the basis of 13
C‐NMR data, they proved that natural
pogostol was neither identical to H nor to its 7‐epimer. However,
they did not propose any alternative structure. The structure of
pogostol was then corrected to I (Figure 4) by Amand et al. on the
basis of renewed NOE observations in two different solvents.244 In
CDCl3 H4 and H5 are separated by only 0.01 ppm complicating
NOE conclusions, which probably led to four groups publishing the
same but wrong structure H. In pyridine, these two atoms are much
better separated. In 2014, pogostol was isolated from a fungus and H
was given as the structure.245 In the 90s we have isolated pogostol
13
from PEO and carried out detailed NMR studies. The C‐NMR data
are identical to those published by Itokawa, Weyerstahl, Stierle,
Amand and Barra. One of the authors (TvB) arrived at the same con-
clusion as Amand et al. concerning the relative stereochemistry.246
However structure I is not in agreement with the evidence provided
by Teisseire. As pogostol can be converted to α‐bulnesene and
bulnesol, 37 represents the correct structure for pogostol, i.e., the
enantiomer of I. Although not being hard evidence, the fact that all
compounds in PEO without double bonds at C4, C5 and C7, like α‐
patchoulene 24, γ‐patchoulene 23, α‐bulnesene 15, seychellene 26
and patchoulol 33, have the same stereochemistry at C4, C5 and
C7 as 37, supports the suggested absolute stereochemistry. Conse-
quently, the structure assigned in the CAS Registry to (−)‐pogostol
with the chemical name (1R,3aS,4R,7S,8aR)‐decahydro‐1,4‐dimethyl‐
7‐(1‐methylethenyl)‐4‐azulenol with CAS RN [21698‐41‐9] is wrong,
as it corresponds to (+)‐ent‐pogostol. The – in our view – correct
structure of pogostol present in PEO is shown in Table 3A, with
the (1S,3aR,4S,7R,8aS)‐configuration, and is therefore pending a new
CAS RN.
3.3.11 | Selinane SQHCs
In considering the fairly large number of selinane (syn. eudesmane)
hydrocarbons identified in PEO, albeit in relatively minor proportions,

VAN BEEK AND JOULAIN
one could expect that traces of oxygenated selinane derivatives
are present as well, but no such sesquiterpenoids have been
identified yet.
3.3.12 | Sesquiterpene ketones
Only a few sesquiterpene ketones have been reported as constituents
156
of patchouli oil, the first one by Teisseire et al. They proposed two
possible structures without stereochemistry, J or K (Figure 5). Later the
spectroscopic data were critically re‐examined and the compound was
identified as α‐bulnesenone.197 α‐Bulnesenone was recently synthe-
sised and its structure confirmed on the basis of EI‐MS and NMR
data.247 However this ketone could not be detected when searching
for ions at m/z 218 (molecular ion, 41% rel. int.) and m/z 108 (base
peak) in the TIC trace from a GC‐MS analysis of PEO (Joulain, unpub-
lished). Thus it remains doubtful whether Teissseire's ketone was actu-
ally α‐bulnesenone. A retrospective correlation is impossible as
1
Teisseire et al. published only UV, IR and H‐NMR data and Givaudan
only 13C‐NMR and EI‐MS data.
Later Mookherjee et al. reported "strong‐woody‐patchouly"
β‐patchoulenone 39, "strong carvone, camphoraceous" α‐guaienone
98 (no relative stereochemistry provided), "strong ambergris"
α‐patchoulone 99, and "odorless" 9‐oxopatchoulol 103 to be present
183
in "traces" in PEO. However as the results have never been
published in a peer‐reviewed journal and nobody has since fully
identified 98, 99 and 103 in PEO, their report should be considered
tentative until they are isolated and confirmed by modern spectro-
scopic techniques. In contrast, however, 39 was already known as a
synthetic product by oxidation of β‐patchoulene,248 which contributed
to its identification. Buré and Sellier detected an ill‐defined “9‐
oxopatchoulol” but without spectroscopic proof its identification
remains tentative.208
FIGURE 5 Structures of discussed sesquiterpene ketones
Despite its patchouli‐related name, (−)‐patchoulenone cannot be
detected in PEO (Joulain, unpublished). However it does occur in
Cyperus species.249-251 Consequently, the compounds that have been
wrongly identified as "patchoulenone" and "isopatchoulenone"
in PEO, 208
still need to be correctly identified. Some of this con-
troversy has been caused by Akhila and Tewari, who in their
review on patchouli oil, incorrectly cited Trivedi et al.35,249 When
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
17
β‐patchoulenone 39, α‐patchoulone 99 and 9‐oxopatchoulol 103 were
cited as PEO constituents,208,252, 99 was already known as a hemi‐
synthetic product.253
As a matter of fact, rotundone 40 is likely to correspond to one of
the aforementioned ketones. It has been repeatedly (inadvertently?)
cited as a constituent of PEO,254,255 although no detailed report has
ever been published on its isolation and unambiguous identification
by well‐accepted spectroscopic techniques. A constituent having the
typical MS and GC LRIs on non‐polar and polar columns of rotundone
can indeed be easily detected in PEO (Joulain, unpublished;
Posthumus, personal communication; see also Figure 7c). The presence
of 40 in PEO is not surprising since it has been shown to be readily pro-
duced by aerial oxidation of α‐guaiene 16.256
Four norsesquiterpene ketones 41‐44 were recently identified as
trace constituents (< 100 ppm) in PEO by Liu et al.160 Although the
starting oil (215 g) was lab‐distilled, one may question the naturalness
of the methoxylated ketone 43, which could well be an artifact formed
by reaction with methanol, which was used as a solvent during the
isolation procedure.
Li et al. published the structures of five hydroxysesquiterpene
ketones, one known one and four new ones 104‐108 (MW= 234
Da) occurring in a 70% ethanolic extract of patchouli stems.159 After
re‐analysing the data of several of our own patchouli analyses, at
least four trace constituents in PEO with a mass spectrum, which
could very well fit the published structures could be detected (Joulain
and Posthumus, unpublished results). Unfortunately, Li et al. did not
publish EI‐MS spectra so for now it is unclear which structure fits
which mass spectrum and which peak in the chromatogram and thus
the five ketones are in Table 3B. The same holds true for the recently
discovered pocahemiketal A 45 and B 46 as new low ppm elements
of PEO.161
3.3.13 | Other oxygen‐containing sesquiterpenoids
The identification of the "strong woody" α‐cedrenal 102 (Table 3B)
cited by Mookherjee without providing proof needs to be confirmed,
since no other cedrane sesquiterpenes have been identified so far in
PEO.183 In contrast, sesquiterpene oxides, such as the guaiene oxides
28/29 and caryophyllene oxide 30 have been fully identified in PEO
a long time ago (Table 3A). It is noteworthy that the former two have

18
been detected recently among the products generated by simple
aerial oxidation of α‐guaiene 16, together with rotundone 40 and
7‐epichabrolidione A 97.256,257 Since 16 is a major constituent of
PEO (~11%, Table 4), it is not surprising to find these same ketones
as well as the C‐1 epimer 101 of 7‐epichabrolidione 100 in PEO
(Joulain, unpublished; Table 3B). The latter compound had already
been identified in a soft coral.258 Air oxidation of α‐guaiene is thought
to proceed through a hydroperoxide acting as a common intermediate
for the production of 28, 29, 40, 100 and 101 as well as an oxidising
agent for the production of other minor oxygenated sesquiterpenes
from other substrates.256,257 The same may be true for hydroperox-
259
ides, which occur naturally in patchouli leaves, but do not exist in
the essential oil since they are likely to be destroyed during steam or
hydrodistillation. Many other minor mono and dihydroxylated sesqui-
terpenes as well as hydroxylated sesquiterpene ethers and ketones
can be detected by GC‐MS in PEO.
Spathulenol, an aromadendrane tertiary alcohol, can
detected sometimes as a trace element in PEO but its presence is
most likely due to adulteration. It is also known to be an artifact
produced by air oxidation of bicyclogermacrene.260 However the
latter compound has so far not been detected in PEO, possibly due
to a co‐elution with major SQHCs on both non‐polar and polar
stationary phases.
3.3.14 | Pogostone
In contrast to patchoulol, pogostone 51 is specific for P. cablin and such
a single species occurrence is a rare feature in the plant kingdom. The
same holds true for its analogs 47‐50 and 52‐53, which occur in much
smaller concentrations in PEO. Pogostone was first identified by Klein
et al.261 The ratio of patchoulol to pogostone varies markedly in PEO,
which may be due to the existence of different chemotypes of P.
262 263
cablin, and/or selective accumulation of pogostone in stems.
recently, two independent research groups have investigated these
hypotheses in more detail and moreover suggested that tissue age
264,265
may also play a role. All these factors together could certainly
account for the high variability of the pogostone content in PEO,
depending of the proportions of stems versus leaves and age of the cul-
tivar used for the production of PEO. Another factor might be related to
the high acidity of 51 (predicted pKa ≈ 6). If PEO contains a significant
percentage of 51, the “acid value” according to titration will be “out of
spec”. To remedy this, such a PEO could be washed with base, which in
turn will contribute to variation in the % of 51. Interestingly, Howarth
reported on a higher acidity of PEOs originating from Sulawesi (acid
value 11‐17) compared to those from Sumatra and Java (acid value 3‐
5) with a concomitant drop in patchoulol content from ~32% to
~27%.10 The lower quality of PEOs from Sulawesi might be caused by
an increased pogostone concentration. There are many recent papers
on the biological activity of pogostone as well as patents on its possible
applications. This topic is beyond the scope of this review, however
several of those references are included in the SI.
3.3.15 | Nitrogen‐containing compounds
About 20 nitrogen‐containing compounds, i.e., alkaloids, can be
detected by GC‐MS when analysing a basic fraction of PEO (Joulain,
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
unpublished data). Bearing in mind that such LLE procedures at high
and low pH may generate artifacts, it is noteworthy that no direct GC
analysis of PEO with an N‐specific or selective detection (NPD or
chemiluminescence, respectively) has ever been carried out. The
major ones contain one nitrogen atom only (odd molecular weight).
Patchouli pyridine 69 and epiguaipyridine (Figure 6) have been
reported as the two main nitrogenous components of PEO.266 The
latter structure was a few years later corrected by van der Gen
who, after synthesis, suggested that the main pyridine sesquiterpene
of PEO has in fact the 7R,10S (cis‐) configuration and is thus
guaipyridine 70.267
be
FIGURE 6 Structure of 10‐epiguaipyridine
In a review of N‐compounds in essential oils, Maurer cited the
original paper by Büchi and consequently drew the epi isomer.268
The 1H NMR data from van der Gen was insufficient to make an
unambiguous identification but recently the structure was definitely
established as 70.251 Although guaiol pyridine or epicananodine 117
has been tentatively identified by NMR, its possible formation as an
artifact by hydration of 70 (or even from patchouli pyridine 69)
during the isolation procedure involving strong acid treatment, should
not be ruled out. As a matter of fact, only guaipyridine 70 can be
Very detected by direct GC‐MS analysis of PEO and patchouli solvent
extracts by extracting ions m/z 214 and 215 (Joulain, unpublished).
Consequently, the presence of 69 in PEO needs to be confirmed.
The other minor N‐containing compounds cited in the litera-
ture185,268 have been withheld from Table 3A. More evidence is
needed prior to their identification as true constituents of PEO
(Maurer, personal communication). As of today, the biogenetic origin
of these sesquiterpene pyridines is unclear.
3.3.16 | Eugenol, benzaldehyde and cinnamic aldehyde
Eugenol 68 was first reported as a minor PEO constituent (8 g from
22 kg; 0.036%) in 1904.269 It was isolated from PEO by a combina-
tion of solvent enrichment, fractional vacuum distillation, extraction
with sodium carbonate (phenolic fraction) and again distillation. It
was characterised by its odour, b.p., density, colouration with Fe(III)
and m.p. of a derivative. Similarly, “traces” of benzaldehyde and
cinnamic aldehyde were detected. They were somewhat less rigor-
ously identified by their smell and a non‐depressed m.p. of a mixture
of derivatives of the isolated products and synthetic benzaldehyde
and cinnamic aldehyde.269 The presence of eugenol was also reported
by Gogröf who used TLC as well as a hybrid of column chromatogra-
phy and thick layer chromatography to separate it from other PEO
constituents.128 A few years later Dummond who studied commercial
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 19

PEOs of various origin, confirmed Gogröf's findings.136 Identification
proceeded by smell, various physical parameters and properties of
several derivatives. The concentration was determined as 0.27%. This
suggests either an analytical error (the author also reported not less
than 1.15% (!) of a basic constituent in PEO, which is highly unlikely)
or a contamination of the starting PEO, since eugenol cannot be
detected by GC‐MS and ion extraction (m/z 164) in any genuine,
lab‐distilled oil investigated by us. However, in using the highly sensi-
tive and selective method for the quantification of fragrance allergens
270
(CEN method), eugenol can indeed be detected, but its content in
genuine oils does not exceed 100 ppm (Joulain, unpublished data).
This confirms somehow previous findings where eugenol is reported
among other traces of phenolics and acids after extraction with
60,271
KOH or as a sub 100 ppm constituent. However as the latter
oil was contaminated, it is impossible to be certain where such traces
originated from. Possibly the eugenol observed by both Gogröf and
Dummond by means of TLC, was pogostone 51 as the two
compounds have the same Rf value in various solvents (Joulain,
unpublished data).
Similar to the 1904 Schimmel report, Dummond also reported
“traces” of benzaldehyde and cinnamaldehyde but neither of them
could be detected by us in a range of PEOs by routine GC‐MS.136,269
Hussin reported benzaldehyde but not cinnamaldehyde as a sub‐100
ppm constituent. Masada reported high concentrations of these three
compounds with eugenol actually being the main constituent (!) of the
oil. This implies an extremely adulterated PEO.272 Morris stated
that eugenol, cinnamaldehyde and benzaldehyde originate from
P. heyneanus,273 but this seems unlikely as Murugan did not detect them
in P. heyneanus oil.181 Also Hollo et al. reported the occurrence of
eugenol and benzaldehyde based onTLC and IR.274 Their proof appears
unsatisfactory and possibly the authors were biased, i.e., based on ear-
lier reports they wanted to see compounds, which were not there. Else
they dealt with heavily adulterated PEO. Until further proof, it was
decided not to enter benzaldehyde and cinnamaldehyde in Table 3B.

TABLE 3A Compounds positively identified in patchouli essential oil

Chemical structure Common Chemical Name MW CAS RN Identification reference

1 α‐pinene 136 80‐56‐8 206

2 β‐pinene 136 127‐91‐3 206

206
3 limonene 136 138‐86‐3

4 pentadecane 212 629‐62‐9 Joulain (unpublished)

Posthumus (unpublished)

5 α‐humulene 204 6753‐98‐6 156

163
6 germacrene D 204 23986‐74‐5

7 β‐elemene 204 515‐13‐9 154

8 δ‐elemene 204 1206‐34‐4 206,247

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
20 VAN BEEK AND JOULAIN

TABLE 3A (Continued)

Chemical structure Common Chemical Name MW CAS RN Identification reference

163
9 selina‐4,11‐diene 204 17627‐30‐4

10 α‐selinene 204 35387‐23‐6 163

11 δ‐selinene = selina‐4,6‐diene 204 473‐14‐4 163

12 β‐selinene 204 473‐12‐1 163

163
13 7‐epi‐α‐selinene 204 123123‐37‐5

163
14 selina‐4(15),7(11)‐diene 204 515‐17‐3

15 α‐bulnesene (= δ‐guaiene) 204 3691‐11‐0 149,152

16 α‐guaiene 204 3691‐12‐1 152

163,207
17 aciphyllene 204 87745‐31‐1

153,154
18 (E)‐β‐caryophyllene 204 87‐44‐5

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 21

TABLE 3A (Continued)

Chemical structure Common Chemical Name MW CAS RN Identification reference

163
19 (E)‐7‐epi‐β‐caryophyllene 204 68832‐35‐9

163
20 (Z)‐β‐caryophyllene 204 118‐65‐0

21 β‐patchoulene 204 514‐51‐2 152

22 δ‐patchoulene 204 53823‐16‐8 156,197

23 γ‐patchoulene 204 508‐55‐4 215

24 α‐patchoulene 204 560‐32‐7 153

25 α‐copaene 204 3856‐25‐5 167

154,196
26 seychellene 204 20085‐93‐2

211
27 cycloseychellene 204 52617‐34‐2

156,275
28 1β,5β‐epoxy‐guai‐11‐ene 220 10224‐59‐6

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
22 VAN BEEK AND JOULAIN

TABLE 3A (Continued)

Chemical structure Common Chemical Name MW CAS RN Identification reference

155,156
29 1α,5α‐epoxyguai‐11‐ene 220 10410‐05‐6

155,156
30 caryophyllene oxide 220 1139‐30‐6

155,156
31 1,10‐epoxy‐guai‐11‐ene 220 133647‐70‐8

32 (Z)‐nerolidol 222 3790‐78‐1 Joulain (unpublished)

Posthumus (unpublished)

151,188
33 patchoulol 222 5986‐55‐0

231,232
34 norpatchoulenol 206 41429‐52‐1

155,162,276
35 nortetracyclopatchoulol 206 not assigned

277
36 6‐hydroxypatchoulol 238 1287235‐52‐2

156,244,246,275
37 pogostol 222 not assigned

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 23

TABLE 3A (Continued)

Chemical structure Common Chemical Name MW CAS RN Identification reference

259
38 guaia‐10(15),11‐dien‐1α‐ol 220 133593‐97‐2

39 β‐patchoulenone 218 37932‐11‐9 183,248

254,255
40 rotundone 218 18374‐76‐0

160
41 14‐nor‐β‐patchoul‐1(5)‐ene‐4‐one 204 93570‐32‐2

160
42 14‐nor‐β‐patchoul‐1(5),2‐diene‐4‐one 202 1899836‐02‐2

160
43 2β‐methoxy‐14‐nor‐β‐patchoul‐ 234 1899832‐98‐4
1(5)‐ene‐4‐one

160
44 14‐nor‐β‐patchoul‐1(5)‐ene‐2,4‐dione 218 1902118‐42‐6

161
45 pocahemiketal A 264 not assigned

161
46 pocahemiketal B 236 not assigned

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
24 VAN BEEK AND JOULAIN

TABLE 3A (Continued)

Chemical structure Common Chemical Name MW CAS RN Identification reference

271
47 4‐hydroxy‐6‐methyl‐3‐(1‐oxobutyl)‐ 196 22073‐85‐4
2H‐pyran‐2‐one

271
48 4‐hydroxy‐6‐methyl‐3‐(2‐methyl‐1‐ 196 22073‐86‐5
oxopropyl)‐2H‐pyran‐2‐one

49 4‐hydroxy‐6‐methyl‐3‐(3‐methyl‐1‐ 210 872310‐80‐0 Joulain (unpublished)

oxobutyl)‐2H‐pyran‐2‐one Posthumus (unpublished)

271,278
50 4‐hydroxy‐6‐methyl‐3‐(2‐methyl‐1‐ 210 58667‐67‐7
oxobutyl)‐2H‐pyran‐2‐one

261,271,278
51 pogostone; dhelwangin 224 Enol : 23800‐56‐8
Keto: 58667‐66‐6

271
52 4‐hydroxy‐6‐methyl‐3‐(1‐oxohexyl)‐ 224 27424‐82‐4
2H‐pyran‐2‐one

271
53 4‐hydroxy‐6‐methyl‐3‐(5‐methyl‐1‐ 224 69793‐67‐5
oxohexyl)‐2H‐pyran‐2‐one

54 nonyl acetate 156 143‐13‐5 Joulain (unpublished)

Posthumus (unpublished)

271
55 2‐methylbutyric acid 102 116‐53‐0

271
56 pentanoic acid 102 109‐52‐4

271
57 4‐methylpentanoic acid 116 646‐07‐1

271
58 2‐methylpentanoic acid 116 97‐61‐0

271
59 heptanoic acid 130 111‐14‐8

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 25

TABLE 3A (Continued)

Chemical structure Common Chemical Name MW CAS RN Identification reference

271
60 nonanoic acid 158 112‐05‐0

271,279
61 cis‐2‐pentylcyclopropanecarboxylic acid 156 5075‐48‐9

271,279
62 trans‐2‐pentylcyclopropanecarboxylic acid 156 58650‐45‐6

271
63 phenol 94 108‐95‐2

271
64 o‐cresol 108 95‐48‐7

271
65 dimethylphenol 154 undefined

271
66 4‐vinylphenol 120 2628‐17‐3

271
67 guaiacol 124 90‐05‐1

271
68 eugenol 164 97‐53‐0

266
69 patchoulipyridine 215 6517‐97‐1

251
70 guaipyridine 215 41447‐48‐7

268
71 (E)‐3‐(but‐1‐enyl)pyridine 133 142505‐11‐1

185
72 (Z)‐3‐(but‐1‐enyl)‐4‐propylpyridine 175 142505‐12‐2
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
26 VAN BEEK AND JOULAIN

TABLE 3B Compounds tentatively identified in patchouli essential oil

Chemical structure Common chemical name MW CAS RN Identification
165
73 2‐ethylfuran 96 3208‐16‐0

74 α‐curcumene = ar‐curcumene 202 644‐30‐4 Joulain (unpublished)

75 η−selinene 204 599‐57‐5 Joulain (unpublished)

76 ε‐patchoulene 204 172925‐72‐3 Joulain (unpublished)

184
77 hotrienol 152 20053‐08‐7

165
78 safranal 150 116‐26‐7

165
79 2‐methylisoborneol 168 2371‐42‐8

36,165
80 geosmin 182 19700‐21‐1

165
81 ethyl 4‐methylpentanoate 144 25415‐67‐2

82 guaia‐1,11‐dien‐10‐ola 220 not assigned 162

83 guai‐11‐en‐5‐ola 222 not assignedb 162

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 27

TABLE 3B (Continued)

Chemical structure Common chemical name MW CAS RN Identification

206
84 (E)‐phytol 296 150‐86‐7

206
85 hexahydrofarnesylacetone 268 502‐69‐2

36
86 4,5‐epithiocaryophyllene 236 65563‐95‐3

36
87 1,2‐epithiohumulene 236 65563‐96‐4

183,252
88 2,3‐dimethyl‐2‐cyclohexen‐1‐one 124 1122‐20‐9

183,252
89 3,4‐dimethyl‐2‐cyclohexen‐1‐one 124 10463‐42‐0

183,252
90 3‐methyl‐2‐cyclohexen‐1‐one 124 1193‐18‐6

183,252
91 isophorone 138 78‐59‐1

183,184,252
92 2,3,4,4‐tetramethyl‐2‐cyclohexen‐1‐one 152 28354‐98‐5

183,252
93 3,5‐dimethylcyclohexan‐1,4‐dione 140 14384‐30‐6

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
28 VAN BEEK AND JOULAIN

TABLE 3B (Continued)

Chemical structure Common chemical name MW CAS RN Identification

183,252
94 4‐oxoisophorone 152 1125‐21‐9

155
95 dihydrobovolide 182 10547‐84‐9

184
96 4‐(3‐methyl‐2‐furyl)‐2‐butanone 152 898543‐80‐1

97 α‐bulnesenone 218 143072‐17‐7 Joulain et al. (unpublished)

98 α‐guaienone 218 not assigned 183,252

99 α‐patchoulone = patchoulione 220 5986‐56‐1 183,253

100 7‐epichabrolidione A 236 1633708‐84‐5 Joulain et al. (unpublished)

101 1,7‐diepichabrolidione A 236 1644642‐00‐1 Joulain et al. (unpublished)

102 α‐cedrenal 218 28587‐62‐4 183,252

183,252
103 9‐oxopatchoulol 236 111931‐60‐3

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 29

TABLE 3B (Continued)

Chemical structure Common chemical name MW CAS RN Identification

159
104 (4R)‐4‐hydroxyrotundone 234 1564275‐90‐6

159
105 (4R)‐4‐hydroxy‐10‐epirotundone 234 190451‐21‐9

159
106 cis‐4‐hydroxy‐β‐patchoulen‐2‐one 234 1564275‐86‐0

159
107 trans‐4‐hydroxy‐β‐patchoulen‐2‐one 234 1564275‐89‐3

159
108 (4R)‐4‐hydroxy‐α‐patchoulen‐8‐one 234 1564278‐84‐8

165
109 4‐ethylphenol 122 123‐07‐9

165
110 3‐propylphenol 136 621‐27‐2

165
111 4‐ethylguaiacol 152 2785‐89‐9

165
112 4‐vinylguaiacol 150 7786‐61‐0

185
113 (E)‐3‐(but‐1‐enyl)‐4‐propylpyridine 175 142505‐13‐3

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
30 VAN BEEK AND JOULAIN

TABLE 3B (Continued)

Chemical structure Common chemical name MW CAS RN Identification

185
114 (Z)‐3‐(but‐1‐enyl)pyridine 133 142505‐10‐0

185
115 5‐isopropyl‐2‐picoline 135 20194‐71‐2

185
116 5,6‐dihydro‐3,6,6‐trimethyl‐7H‐ 175 55713‐43‐4
cyclopenta[c]pyridin‐7‐one

251
117 guaiol pyridine = epicananodine 233 41447‐00‐1

183,252
118 dehydroguaiol pyridine 231 not assigned

185
119 6,7‐dihydro‐1,3,6,6‐tetramethyl‐ 203 55713‐38‐7
8(5H)‐isoquinolinone

185
120 122 undefined

185
121 136 undefined

185
122 150 undefined

185
123 164 undefined

165
124 2‐methoxy‐3,6‐dimethylpyrazine 138 19846‐22‐1

(Continues)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 31

TABLE 3B (Continued)

Chemical structure Common chemical name MW CAS RN Identification

185
125 2‐methoxy‐3‐isopropylpyrazine 152 25773‐40‐4

165
126 2‐isobutyl‐3‐methoxypyrazine 166 24683‐00‐9

185
127 quinoxaline 130 91‐19‐0

185
128 2‐aminoacetophenone 135 551‐93‐9

185
129 methyl anthranilate 151 134‐20‐3

185
130 methyl N‐methylanthranilate 165 85‐91‐6

a
undefined stereochemistry; bCAS RN 1632300‐50‐5 has been assigned to a defined isomer of synthetic origin.256

3.4 | Qualitative and quantitative variation of major hydrocarbons will be correspondingly low and vice versa. However, the
constituents ratios between the hydrocarbons appear to be fairly constant (see § 3.5
on Fingerprints). Besides the natural chemical variation, the drying and
The second aim was to map the variation reported in the literature
distillation process (hydrodistillation versus steam distillation, duration,
among the major constituents of PEO formulated in § 3.2. All reports
pH of still, etc.) will contribute to the variation. The most variable com-
deemed reliable were analysed with respect to the main constituents.
pound of all is the non‐terpenoid pogostone. It can be undetectable
If authors analysed multiple oils, the average % was used. When
(Figure 7a) or be the major constituent (Figure 7b). According to Luo
authors reported a combined % for two or more constituents, such
et al. there are patchoulol and pogostone chemotypes of P. cablin.262
data was not used. When authors did not report the occurrence of a
The same group also reported that stem oil contained twenty times
particular compound, it was not assumed to be zero but rather was
more pogostone than leaf oil, which could also partially explain the
ignored. When authors reported a trace, arbitrarily it was counted as
concentration differences.263 Hu in a fingerprinting paper based on
0.02%. Compounds mentioned in less than eight analyses were
18 PEOs reported patchoulol‐pogostone ratios varying from 0.70 to
disregarded so many trace components mentioned in Table 3A do
9.50.307 Liu et al. reported pogostone concentrations from < 5% to >
not figure in Table 4. Over 100 oil analyses from 75 papers were
65%.308 The average concentration of 8.9% in Table 4 is an exaggera-
evaluated and the results expressed as average percentage and ranges
tion. This is caused by the fact that in many analyses pogostone
are summarized in Table 4. No analytical results by the authors are
occurred at < 1% and was therefore ignored. However, we have not
incorporated in Table 4.
entered 0% each time a compound was not mentioned.
The ranges in Table 4 indicate an enormous variation between oils
However even the wide ranges inTable 4 already give an indication
and of course significant variation is to be expected. In spite of the fact
which compounds are major constituents and which ones are interme-
that, where possible, the dataset has been scrutinized and corrected, it
diate or minor constituents. This is more easily seen from the average
will still contain errors and this is probably most true for the more values, which in our opinion are indeed representative of the average
extreme values. The large ranges are partially due to the widely variable PEO. The total excluding pogostone adds up to ~91% and this fits our
ratio of hydrocarbons to oxygenated constituents in PEO. If a patchouli own observations that the combined constituents in Table 4 together
oil contains 80% of oxygenated compounds, the relative amounts of all with γ and δ‐patchoulene but excluding pogostone, make up 90‐97%
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
32 VAN BEEK AND JOULAIN

TABLE 4 Average concentration (GC, internal normalisation) and
ranges of major patchouli constituents based on over 100 patchouli oil
analyses. Compounds are tabulated in order of elution on a
polydimethylsiloxane phase36,40,58,60-67,69-72,74-76,81,83,86,91,92,99,101,
102,104,106,108,111-113,118-124,129,179,181,182,186,201,206,208,263,280-306
Constituent Average % Ranges % Nr of data
and unique for PEO, are absent as well as the important aroma contributor
nortetracyclopatchoulol. α‐Gurjunene is an indicator of adulteration and
should not be allowed to occur in PEOs, not even at 0.2% in oils prepared
in an artisanal way. The Indonesian norm SNI 06‐2385‐2006 demands a
minimum of 30% patchoulol and a maximum of 0.5% α‐copaene.141

α‐Pinene 0.09 0.01‐0.3 28

TABLE 5 Allowed ranges of several constituents of patchouli oil
β‐Pinene 0.20 0.02‐1 30
according to 2003 ISO norms167
Limonene 0.03 0.01‐0.3 22
δ‐Elemene 0.52 0.01‐1.9 19 Constituent Minimum % Maximum %

β‐Patchoulene 3.0 0.03‐12 78 β‐Patchoulene 1.8 3.5

β‐Elemene 0.89 0.18‐1.9 40 Copaene traces 1
Cycloseychellene 0.46 0.02‐0.8 21 α‐Gurjunene 0.0 0.2a
(E)‐β‐Caryophyllene 3.1 0.75‐6.8 78 α‐Guaiene 11 16
α‐Guaiene 11 2.9‐23 64 β‐Caryophyllene 2 5
Seychellene 6.6 2.3‐13 67 Bulnesene 13 21
α‐Humulene 0.69 0.05‐2 26 Norpatchoulenol 0.35 1
α‐Patchoulene 4.5 1.2‐13 70 Patchoulol 27 35
Germacrene D 0.12 0.0‐0.2 8 Pogostol 1 2.5
Aciphyllene 2.4 0.7‐4.2 30 Total 56.15 85.2
α‐Bulnesene 14 2.9‐23 83 a
only for oils prepared in an artisanal way.
Norpatchoulenol 0.86 0.11‐4.0 37
Caryophyllene oxide 0.73 0.0‐4.6 30
Pogostol 2.4 0.2‐6.2 50 3.5 | Chromatographic fingerprint of patchouli oil
Patchoulol 39 11‐72 104 A gas chromatogram of an essential oil can serve as a fingerprint and
Total without pogostone 90.6 is a powerful tool in controlling the quality of produced or bought
Pogostone 8.9 0.1‐27.7 24 oil.311 Moreover it can show also peaks, which should not be there,
Total with pogostone 99.5 ‐ ‐ i.e., volatile adulterants or impurities. Published, correctly assigned
chromatograms are also helpful for future researchers. Notwithstand-
of PEO. γ and δ‐patchoulene are the only intermediate constituents not ing the wide variation in concentration of individual constituents,
present inTable 4 for the reason that insufficient reliable data was avail- patchouli oil does have a unique profile. In Figure 7a‐c the sesquiter-
able in literature. Interestingly, Schmidt has analysed 52 PEOs from pene regions of the chromatograms on DB5 stationary phase of three
Indonesia, India, China, Madagascar and Brazil over a period of 15 very different PEOs are depicted. Oil A almost meets the ISO norms
years.309 Unfortunately, neither analytical data nor averages are given, and contains no pogostone. Stem oil B contains pogostone in over
only the ranges for 22 constituents. However, if the median of the 20%, some α and β‐pinene and has a very high norpatchoulenol con-
ranges is taken as a kind of average, there is actually a very good corre- tent. Oil C has a very high patchoulol content, some pogostone, several
spondence with the data in Table 4. β‐Elemene is not reported by him
more oxygenated, higher boiling constituents and a low hydrocarbon
but as the concentration reported by Schmidt for α‐copaene is too high
content. The sesquiterpene hydrocarbon pattern is specific with
(0.2‐1.6%; normally ≤ 0.1%) and otherwise the data look reliable, we
relatively low concentrations of δ and β‐elemene, cycloseychellene
assume α‐copaene has been misidentified, even more so as our range
and humulene, intermediate concentrations of β‐patchoulene,
for β‐elemene in Table 4 (0.2‐1.9%) is very similar to his range reported
β‐caryophyllene, α‐patchoulene, δ‐patchoulene and aciphyllene and
for α‐copaene. Rather surprisingly no pogostone is reported. The only
real mismatch is the too high concentration reported by him for high concentrations of α‐guaiene, seychellene and α‐bulnesene.
γ‐patchoulene (5.9‐8.0%). This must be a misidentification although it Compounds specific for PEO include cycloseychellene 27,
is unclear what it was in reality as all major PEO constituents are norpatchoulenol 34, nortetracyclopatchoulol 35, several patchouli
accounted for in Schmidt's table.309 ketones, pogostone 51 and the pyridines 69 and 70. Only the first
When the average percentages and the ranges are compared with the three should always be detectable. Cycloseychellene is easier sepa-
ISO norms, the ISO norms appear to be somewhat restrictive. For instance, rated from (E)‐β‐caryophyllene 18 on a DB1 (Figure S2 in SI) than a
none of the three PEOs depicted in Figure 7a‐c would meet the standard DB5 stationary phase (Figure 7b) but can also be resolved from 18
based on the patchoulol content. If sellers would need to meet all the on DB5 with a different temperature gradient (Figure S4 in SI). The
demands inTable 5 for all their oils, it would require considerable blending patchoulenes, seychellene, patchoulol and pogostol are highly charac-
or manipulation, and indeed blending of premium high‐patchoulol Indone- teristic of PEO. Non‐sesquiterpenoids, which are almost always
sian PEO with cheaper low‐patchoulol Chinese PEO occurs.310 Also the detectable in low concentrations in PEO include α and β‐pinene, nonyl
choice of compounds is odd, copaene is present at trace levels in PEO acetate and pentadecane. The PEO fingerprint is characterised by the
and not typical for PEO and cycloseychellene would be a better choice. combination of the above compounds in relatively fixed relative ratios
Seychellene and pogostone, which are respectively highly characteristic (see Table 6 and Figure 7a‐c).
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 33

TABLE 6 Relative percentages of 15 major constituents of the 3 patchouli oils depicted in Figure 7a‐c
Constituent RI CPSil 5a177 RI DB1b RI DB5176 RI DB5c Oil A Oil B Oil C

α‐Pinene ‐ ‐ 932 ‐ 0.1 1.9 0.0

β‐Pinene ‐ ‐ 974 ‐ 0.3 4.8 0.0
β‐Patchoulene 1388 1384 1379 1388 1.7 2.2 0.2
(E)‐β‐Caryophyllene 1421 1416 1417 1425 3.9 2.1 0.5
α‐Guaiene 1440 1434 1437 1442 13.0 7.5 2.3
Seychellene 1447 1442 1444 1448 7.4 4.3 3.0
α‐Patchoulene 1467 1458 1454 1461 5.3 2.5 1.7
δ‐Patchoulene 1466 1460 ‐ 1465 3.1 1.3 0.6
Aciphyllene 1495 1495 1501 1502 3.1 1.5 0.9
α‐Bulnesene 1503 1502 1509 1510 16.2 10.1 4.2
Norpatchoulenol 1551 1544 1553 1559 1.1 4.5 1.0
Caryophyllene oxide 1578 1572 1582 1575 0.1 0.0 1.0
Pogostol 1647 1641 1651 1656 2.1 2.0 4.3
Patchoulol 1661 1651 1656 1661 36.8 22.9 59.4
Pogostone ‐ 1677 ‐ 1716 0.0 22.0 3.4
Total 94.2 89.6 82.5
a
equivalent to DB1 (dimethyl polysiloxane).
b
van Beek, unpublished; for conditions, see SI, Figure S2.
c
van Beek, unpublished; for conditions, see SI, Figure S4.

FIGURE 7A Partial gas chromatogram (sesquiterpene region) of patchouli oil A on a DB5 stationary phase. Legends: δE = δ‐Elemene, βE = β‐
Elemene, βPa = β‐Patchoulene, βCa = (E)‐β‐Caryophyllene, CyS = Cycloseychellene, αGu = α‐Guiaene, Sey = Seychellene, αHu = α‐Humulene,
αPa = α‐Patchoulene, δPa = δ‐Patchoulene, U = Unidentified constituents, GD = Germacrene D, δSe = δ‐Selinene = Selina‐4,6‐diene, 15 =
Pentadecane, Aci = Aciphyllene, αBu = α‐Bulnesene = δ‐Guiaene, 7eS = 7‐epi‐α‐Selinene, ZNe = (Z)‐Nerolidol, eBu = 1,10‐epoxy‐α‐Bulnesene, nPa
= Norpatchoulenol, Cao = Caryophyllene oxide, ntP = Nortetracyclopatchoulol, Pgl = Pogostol, Pat = Patchoulol. GC conditions: 60 m × 0.25 mm ×
0.25 μm DB5 column; He 18 psi; 60 °C (0 min hold) to 260 °C at 3 °C/min; MS detection (M.A. Posthumus, unpublished results)
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
34 VAN BEEK AND JOULAIN

FIGURE 7B Partial gas chromatogram (sesquiterpene region plus pogostone) of PEO B prepared from stems on a DB5 stationary phase. For peak
assignments and conditions, see Figure 7a. 2mP = 4‐Hydroxy‐6‐methyl‐3‐(2‐methyl‐1‐oxobutyl)‐2H‐pyran‐2‐one 50, 6mP = 4‐Hydroxy‐6‐methyl‐
3‐(3‐methyl‐1‐oxobutyl)‐2H‐pyran‐2‐one 49, Pgn = Pogostone (M.A. Posthumus, unpublished results)

FIGURE 7C Partial gas chromatogram (sesquiterpene region) of a significantly oxidised PEO C on a DB5 stationary phase. For peak assignments
and conditions, see Figure 7a. Rtd = rotundone, 234 = unidentified constituent with MW = 234, possibly identical with 104 ‐ 108 (M.A. Posthumus,
unpublished results)

VAN BEEK AND JOULAIN
3.6 | Quantitative Analysis of Patchoulol
The combined data from MS (to ensure identification) and FID (for
quantification) will provide a fingerprint as well as relative percentages
of the constituents (100% internal normalisation with equal response
factors for all components). If the PEO is genuine, freshly distilled
and properly stored, it will contain only volatiles and thus the found
relative percentages are representative of the entire sample. When
using a flame ionisation detector (FID), hydrocarbons give higher
responses than oxygenated components and thus in PEO the found
percentages for sesquiterpene hydrocarbons are expected to be
slightly too high and that of patchoulol slightly too low, e.g., in reality
33 instead of 30% patchoulol. More accurate figures can be obtained
by using earlier experimentally determined relative response factors
312-314
(RRFs) or predicted RRFs. When using the Total Ion Current
(TIC) of a mass spectrometer as detector, the deviation can be larger
than 10%.315 In the past a thermal conductivity detector (TCD;
katharometer) has also been used for PEO and the author claims that
RRFs do not differ much from 1.316 Van Beek has compared 3 different
detectors on five different GC systems (TCD; MS‐TIC; FID; FID; FID)
for the analysis of patchoulol in PEO by means of internal
standardisation and found absolute percentages of 31.0, 30.6, 30.9,
30.7 and 30.9%.317 At least for patchoulol, this suggests that with
internal standardisation the detector does not strongly influence the
outcome as long as all other parameters are optimal. When PEO is
adulterated with non‐volatile additives, the measured relative percent-
ages will not change but no longer reflect the true absolute percent-
ages. The same holds true when essential oils are old and have been
improperly stored as this leads to non‐volatile polymers.318
The International Organization of the Flavor Industry (IOFI) has
published guidelines for quantitative GC analysis of essential oils.319
For obtaining truly quantitative data, they advise to use external
standardisation, internal standardisation or standard addition. The
use of an autosampler is desirable and to avoid discrimination splitless
or on‐column injection preferred. However even then, quantitative
essential oil analysis remains a complex task with many potential
pitfalls like linearity, widely different concentrations of constituents, loss
or decomposition of analytes during their analysis, non‐availability of
reference compounds, finding a proper internal standard, optimisation
of the many different GC parameters, baseline separation of analyte
peaks, column selection, choice of detector, and, injection volume and
315,318-320
split ratio in case of split injections. The absolute purity of
reference analytes for preparing calibration solutions is an often
neglected issue. Indications from suppliers cannot always be relied
on and a 100% normalisation GC analysis of the standard ignores
non‐volatile impurities. Quantitative NMR against a suitable, non‐volatile,
stable and commercially available reference compound of known
absolute and high purity is the preferred solution.321
For PEO three main aroma volatiles (see section on Odour)
patchoulol 33, norpatchoulenol 34 and nortetracyclopatchoulol 35
are most relevant for an assessment of their absolute concentration.
Patchoulol is commercially available but the other two constituents
are not. However, comparing the well resolved peak areas of 34 and
35 with that of patchoulol should yield also their true concentrations.
Being all three tertiary alcohols, it can be assumed that their RRFs
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
35
are approximately the same when FID is used. Apart from this, there
should be an indirect check on the gross presence of non‐volatile or
extraneous compounds. This is most easily achieved by comparing
the total peak area of all patchouli‐related peaks (see section on Fin-
gerprint) with that of the patchoulol peak with or without correction
of RRFs. Teisseire has discussed an alternative approach.239 Finally
for satisfying the ISO norms, the absolute concentration of several
additional constituents like α‐guaiene 16, α‐bulnesene 15 and pogostol
37 needs to be determined, which again can be done starting from the
absolute concentration of patchoulol 33. Preferably predicted RRFs
should be used in this case.
Surveying the literature, there are quantitative papers on
patchoulol and pogostone only. Both internal60,171,265,317,322,323 and
external standardisation68,89,170,307,324-327 have been applied.
The very first quantitative method for patchoulol was developed
by a committee of experts and used 1‐pentadecanol as internal stan-
dard (i.s.).322 This i.s. elutes shortly after patchoulol and, being a stable
C15 alcohol, is a fair choice, its main disadvantage being the b.p., which
is 57 °C higher than that of patchoulol and additionally it is a primary
alcohol whereas patchoulol is tertiary. Both packed and capillary col-
umns were used. 0.5 μL (10% PEO in ethyl acetate) was injected with
a split of 1:200. The injection temperature was not specified. Detection
was by FID. The purity of patchoulol reference was only determined by
100% normalisation. Van Beek used (E,E)‐farnesol as i.s.317 This com-
pound is well separated from both patchoulol and other PEO constitu-
ents, its b.p. is only 4 °C lower than that of patchoulol and it has the
same oxidation status and elemental composition (C15H26O). Unfortu-
nately, it is not a tertiary alcohol. Cedrol, a saturated tricyclic teriary
alcohol with a b.p. 15 °C lower than that of patchoulol, might be a good
alternative i.s. A GC equipped with one injector (split 1:100) and two 60
m columns (DB1 and DB‐Wax) each with their own FID was used thus
generating 2 chromatograms per injection. Two different PEOs were
analysed in 2008 and 2010. The patchoulol percentages found were
for oil 1 30.9% (DB1, 2008), 30.3% (DB1, 2010), 30.7% (Wax, 2008)
and 30.6% (Wax 2010). For oil 2 these values were: 50.9% (DB1,
2008), 50.2% (DB1, 2010), 50.4% (Wax, 2008) and 50.9% (Wax
2010). The actual percentages are unknown but the results provide
confidence. The method was partially validated by analysing an artificial
PEO consisting of ~10% dodecanol, ~50% tetradecane and 37.36%
patchoulol. Analysis by the same i.s. method yielded 37.26% and
37.40% for the DB1 and DB‐Wax column respectively.317 Details were
unfortunately never published.
Hussin et al. published a quantitative method, which is also lacking in
details. It appears that no true relative response factor (RRF) against the
used i.s. was calculated for patchoulol but rather the average RRF of sev-
eral monoterpene alcohols against the i.s. using FID. Whether this is
appropriate was not validated. As i.s. nonane was used, whose chemical
and physical properties are significantly different from those of
patchoulol. Filippi et al. explicitly mentioned that nonane gives
relatively high fluctuations as an i.s. in quantitative vetiver analyses com-
pared to a higher boiling i.s.318 Nonane does not satisfy several of the
demands specified by the IOFI Working Group on Methods of
Analysis.319 Injection temperature was 280 °C, split ratio 1:100 (10%
PEO in hexane). The injection volume was not specified.60 The fourth
internal standard method was not applied to PEO but rather to

36
headspace samples.323 As i.s. nonyl acetate 54 was used, which is an
unfortunate choice as this is the only simple ester naturally occurring in
PEO, albeit in low concentrations. The last two methods both
used octadecane as i.s. 171,265
Its b.p. is 30 °C higher than that of
patchoulol, which could lead to discrimination during injections. Also its
RRF will differ significantly from that of patchoulol making it altogether
not the ideal i.s. In the Chinese Pharmacopoeia Monograph, the method
was applied to a chloroform extract of P. cablin leaves. The injector tem-
perature was not specified and 1 μL (extract in hexane) was injected with
171
a split of 1:20. The detector temperature was 280 °C. In the last
method the injector temperature was 230 °C and 1 μL (5% PEO in hex-
ane) was injected with a split of 1:50. 265
Overall the fully published
methods do not appear superior to the method using 1‐pentadecanol
published 30 years ago. 322
Still a well‐validated internal standard method
for patchoulol in PEO remains to be published.
The methods using external standardisation have been applied
170,171 307,324,325
to PEO, on patchouli leaves, or on isolated
patchoulol.326,327 All methods are similar in the set‐up of the calibra-
tion curve although the range of concentrations (factor 8 to 1000)
and the number of points (4 to 10) varies. All curves show good
linearity (r2 > 0.995) indicating that nowadays GC autosamplers can
reproducibly inject 1 μL volumes with an RSD of 1%. The injection
conditions vary considerably with temperatures ranging from as low
as 210 °C,325 to as high as 280 °C.324,325 The injection volumes were
1 or 2 μL and split 1:10, splitless or pulsed splitless were used.
Detection proceeded by FID, CI‐MS (monitoring of m/z 221 and 205)
or EI‐MS. In the latter case, TIC, SIM or MS/MS was used. Several
assays were (partially) validated. The best‐validated assay appears the
recent one by Yang et al.89 Details on linearity, recovery (accuracy),
precision, LOD, LOQ, repeatability, stability, inter and intraday
variability are provided by the authors. Some methods also simulta-
neously determine pogostone by GC.89,307 Wang et al. published a
328
GC‐MS method for pogostone only. Octacosane was used as i.s.
Somewhat surprisingly, the authors deemed it necessary to purify
pogostone from PEO prior to GC‐MS by means of two LLE steps.
Due to the interest in its pharmacological properties, in recent years
several LC‐MS methods for the determination of pogostone in
experimental animals have been published. These are not discussed
here; for references, see SI.
3.7 | GC‐IR, Two‐dimensional GC and Chiral GC
3.7.1 | GC‐IR
In the 80s GC‐FTIR was “in trend” and there are two applications in the
field of PEO.163,329 Although FT‐IR is valuable in structure elucidation
and can sometimes provide answers GC‐MS cannot (in the same
paper,329 there are nice examples for distinguishing different santalols
from sandalwood oil by IR), its added value over GC‐MS in the case of
PEO was not demonstrated. Since then GC‐FTIR has faded out.
3.7.2 | Two‐dimensional GC
In principle comprehensive 2‐dimensional GC (GC×GC) is a very
powerful technique for resolving complex essential oils. It can separate
many more components than 1‐dim. GC and thus provide cleaner mass
spectra, especially of minor components. So far GC×GC was applied
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
thrice to PEO.330-332 The study by Ruan is actually inferior to routine
1‐dim. GC of PEO. This is partially caused by using a DB1 column of
only 2 m length in the 1st dim, which on top has the unusual diameter
and film thickness of 100 μm and 3.5 μm respectively. The 2nd dim.
OV17 column is somewhat non‐polar, which does not benefit an opti-
mal resolution. The modulation time was 4 sec. The 2D‐fingerprint
shows some additional minor components but eventually only five
components got identified.330
The second study is more elaborate and three different column
combinations were compared. For their PEO sample the authors
preferred a 60 m Wax column (0.25 mm × 0.25 μm) in the 1st dim.
and a 3 m chiral (Cyclodex‐B) column (0.10 mm × 0.10 μm) in the 2nd
dim. The analytical rationale for choosing a chiral column was that
“the chiral column is a good choice for separating the isomeric compo-
nents in volatile oils of traditional Chinese medicines”. Although it is not
yet known (vide infra) whether mixtures of enantiomers ("scalemic
mixtures") occur in PEO, it is unlikely that any enantiomers can be
resolved within 5 sec. If any constituents would occur as a mixture of
enantiomers, a partial separation would lead to band broadening in
the 2nd dim., which is actually undesirable. Of course the selectivity of
the chiral phase is quite different from the 1st dim. Wax phase and
based on the 2D FID contour plots the combination works well. The
modulation time was 5 sec and a temperature gradient of 70 to
200 °C at 3 °C/min was used. As both columns were present in the
same oven, no individual optimisation of the temperature gradients of
the two columns was possible. Nevertheless, in this study many more
peaks were resolved but clustered classes of compounds (e.g., sesqui-
terpene hydrocarbons and alcohols) have not been indicated. The used
PEO looks highly atypical with many monoterpenes not found in other
studies as well as few sesquiterpene hydrocarbons suggesting contam-
ination or adulteration. The identification of many sesquiterpenes
appears dubious and several characteristic ones like seychellene and
pogostol are suspiciously absent. The authors claimed that they
“tentatively” identified 394 compounds. Relative concentrations are
not given. Overall this study was unfortunately not the breakthrough
in PEO analysis.
Edwards et al. used 20 m BPX5 column (0.18 mm × 0.18 μm) in
the 1st dim. and a 2 m DB1701 (= 14%‐cyanopropyl‐phenyl)‐
methylpolysiloxane) column (0.25 mm × 0.15 μm) in the 2nd dim.332
Detection was by EI‐TOF‐MS at both 70 and 12 eV. As this was only
an exploratory study with PEO being one of 8 analysed essential oils,
it was not in‐depth and only patchoulol is reported. However, all
the major and intermediate PEO constituents could be identified
(McGregor, personal communication). The hardware is fine but the
contour plot shows little added value over a proper 1‐dim. GC study
on a 60 m column. This is caused by a limited difference in selectivity
between the BPX5 and DB1701 columns. A combination of a DB5
and a Wax column would probably have shown more resolved peaks.
Thus a good GC×GC study on PEO, comparable to the state‐
of‐the‐art one on vetiver oil,318 is regrettably still lacking. Anyhow,
even if such a study would be performed, it is doubtful whether it
would lead to many newly identified components as the mass spectral
identification is only as good as the mass spectral database and many
PEO components probably do not occur in the commercial databases.
However due to the much higher chromatographic resolution of

VAN BEEK AND JOULAIN
the GC×GC separation, at least one would know how many more
constituents are awaiting their structural elucidation, i.e., they would
be upgraded from “unknown unknowns” to “known unknowns”.
3.7.3 | Chiral GC
Chiral phases for GC can be used for two different purposes: (1) for
true chiral GC for the distinction of enantiomers or (2) as a stationary
phase with a unique selectivity independently of chirospecific
separation purposes. The latter application can be used preparatively
333
too. Betts used chiral GC (Chiralsil‐Val phase) for purpose (2), i.e.,
to achieve a better separation of patchouli hydrocarbons than on
standard, e.g., dimethyl polysiloxane, stationary phases. However
according to today's standards, the resolution does not appear superior
to chromatograms currently obtained on DB5 stationary phases. An
advantage though is that with 20 min the separation was rather
fast.334 Wu used a Cyclodex‐B column as the 2nd dim. column in a
331
GC×GC application (vide supra). Thus the first true chiral GC study
of PEO, e.g., of purified constituents or after heartcutting, still has to
take place. Based on reported optical rotations for patchoulol and
patchoulenes and biosynthetic considerations, it is expected that these
PEO compounds do not occur as mixtures of enantiomers but this is
still to be proven.
4 | A D U L TE R A T I O N A N D CO NT A M I N A T I O N
335
Different adulteration practices exist in the field of essential oils.
The exact form depends mainly on the chemical composition and the
value of the targeted essential oil. Adulteration, that is, intentional
alteration or falsification, can occur in the case of PEO: (1) prior to
distillation by adding extraneous materials to genuine patchouli leaves;
or (2) after distillation by "supplementing", i.e., diluting PEO with
various ingredients. Deliberate adulteration should be distinguished
from inadvertent contamination as analytically they are quite
different to detect. For adulteration to be economically worthwhile,
the addition of at least 20% of adulterant is necessary, resulting in
significant changes in physical parameters, constituents and/or odour.
In contrast, contamination with a few % of leaves from another species
or soil during steam distillation will generally be more difficult to
detect. Some forms of PEO adulteration, which used to be common,
are obsolete today thanks to the tremendous progress in analytical
chemistry.
Adulteration of PEO has occurred for a long time. The addition of
leaves from other, cheaper plant species to "Java patchouli" was cited
already in the 19th century.17 The same goes for post‐distillation
adulteration of PEO: when pure, the latter was said to dissolve in an equal
bulk of 95% ethanol; if not, adulteration with copaiba balsam should be
suspected.336 Later, the frequent adulteration of PEO was mentioned
several times,24 including in dedicated textbooks on essential oils.26
Over time many different materials have been added – up to 80%
– to genuine patchouli leaves prior to distillation. Ocimum basilicum var.
pilosum, Urena lobata var. sinuate, Hyptis suaveolens, Plectranthus
fruticosus, Lavatera olbia, Pogostemon heyneanus, Microtoena insuavis,
or Pavonia weldenii and even sand or soil – up to 50% – or water –
up to 35% – to increase the overall weight, have been mentioned. 27,48
Substances added to PEO after distillation, can be either natural (cedar
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
37
oil higher fractions, pepper oil, white camphor oil, guaiac wood oil,
copaiba and gurjun balsams, residues from vetiver and camphor oil
productions, cubeba oil, amyris oil) or synthetic (vide infra).27,48,295,335
In the past adulteration of patchouli resinoid oil with oakmoss
resinoids, with cloves and with coumarin, or even diluted with PEO
has been described.173
Many different synthetics have been used to adulterate PEO. At
least 20% of phthalates in a PEO of French origin has been reported
one century ago.337 The authors remarked that “The prices paid at
present for pure patchouli oil make such manipulations exceedingly
profitable”. The addition of glycols such as hexylene glycol, propylene
glycol and dipropylene glycol, benzyl benzoate, various phthalates or
methyl esters of hydrogenated resin acids (Hercolyn D) as well as
non‐volatile adulterants such as vegetable oils (e.g., palm oil), fixed oils
(e.g., castor oil) or mineral oil has been cited.35,48,75,186 Non‐volatile
adulterants are not detectable by conventional GC. However a
straightforward method to detect and quantify those, is integration
of the 15‐20 most intense peaks in GC and comparison with an
internal standard (“semi‐quantification”, see also § 3.6).315,322 The
study by Bruns is particularly valuable as it provides GC profiles of
PEO spiked with gurjun balsam and it shows where various glycols,
isobornyl acetate and (Z)‐8‐camphene methanol (syn. (Z)‐patchouli
ethanol) elute.75 More recently, a paper using a PEO adulterated
with 14.3% methyl benzoate appeared.338 Most of these adulteration
cases would have been detected by an odour assessment by
trained evaluators.
Until the introduction of modern analytical techniques in the
second half the 20th century, colour tests, standard physical measure-
ments (mostly solubility tests, density, refractive index, etc.) and
chemical determinations such as the saponification number, were used
to detect gross falsifications, albeit with probably limited efficacy.25
The first report on the use of gas chromatography to detect the
fraudulent addition of the much less expensive gurjun balsams
(Dipterocarpus spp.) to PEO was published in 1966,153 and the efficacy
of this technique was rapidly confirmed by others.339 However initially
this did not lower the frequency of adulteration.90 Nowadays GC
fingerprinting based on GC‐FID or GC‐TIC chromatograms (see § 3.5)
is the number one quality control method to detect abnormalities in
PEO. Other less used QC techniques capable of detecting adulteration
13
include C‐NMR295 and HPTLC.340 In contrast with some other
economically important essential oils, stable isotope mass spectrome-
try has not yet been used to detect possible adulteration of
PEO constituents.
Over time adulteration of PEO with gurjun and copaiba balsams
was most prevalent and therefore received a lot of attention.75,153,339
α‐Gurjunene (Figure 8) does not occur in PEO and has long
been recognised as an indicator for the presence of gurjun
balsam.75,153,339,341 Recently, the presence of alloaromadendrene
(Figure 8) in PEO was addressed in detail and it was confirmed that
its detection is also indicative of the addition of gurjun balsam.205,206
Unfortunately in the same year a committee of analytical experts
misidentified δ‐patchoulene 22 as alloaromadendrene in PEO and thus
caused some confusion.179 The detection of calarene (Figure 8), a
constituent of Dipterocarpus jourdainii and D. intricatus balsams,339
was not mentioned by Cornwell.206 In summary, neither α‐gurjunene

38
nor alloaromadendrene or other aromadendrane‐type sesquiterpenes
such as spathulenol (Figure 8) should be considered as constituents
of genuine PEO. These aromadendrene‐type sesquiterpene hydrocar-
bons are absent in another gurjun balsam (Dipterocarpus costatus),
which contains mainly (E)‐β‐caryophyllene and α‐humulene.339 The
addition of this balsam can be detected (and also copaiba balsam, for
the same reason) since these two sesquiterpene hydrocarbons occur
75
in lower relative percentages in genuine PEO.
FIGURE 8 Structures of alloaromadendrene, α‐gurjunene, spathulenol and calarene
Some contamination of PEO by crop protection agents or resi-
dues in the still from earlier distillations is possible.13 To our knowl-
edge, no report has been published to date on the detection and
quantification of pesticides in PEO. Various phytosanitary treat-
ments are currently involved during the cultivation of patchouli to
counteract diseases caused by viruses, bacteria, fungi and other
pests.48
Adulteration of PEOs, which are sold to major fragrance houses
appears less prevalent today as the biggest end users of this natural
product have the analytical expertise to detect it. However PEOs sold
in small bottles in health or “well‐being” shops, for e.g., aromatherapy,
are still frequently adulterated (van Beek, unpublished). A chromato-
gram of a PEO bought commercially in The Netherlands containing
5% benzyl alcohol, 22% α‐gurjunene, 3% thujopsene, 7%
alloaromadendrene, 2% γ‐gurjunene, 3% cedrol and 20% benzyl
benzoate, i.e., 1 part PEO and 2 parts adulterants (!), is shown in the
SI. Another example is the oil used in a study on antimicrobial activity
of essential oils.342 It contained 11.3% α‐gurjunene, 6.7% benzyl
alcohol, 3.1% thujopsene, 2.8% aromadendrene and 2.6% α‐cedrol,
which at the same time casts doubts on the antimicrobial results.342
In 2017 a PEO containing 17.8% aromadendrene was used to
determine the effect of PEO on human skin cells.343
Although it has not yet been reported, theoretically the
commercial availability of Clearwood™, a potential patchouli substitute
(see § 5.2), which is 25% cheaper per kg than "regular" PEO, creates an
opportunity to adulterate genuine PEO. Such adulteration will be
harder to detect as Clearwood™ consists of the usual patchouli
SQHCs, pogostol and patchoulol, i.e., all compounds normally present
in PEO. However, Clearwood™ contains patchoulol ethyl ether in a
significant amount and this compound does not occur in PEO. Thus,
addition of a few % of Clearwood™ to genuine PEO will be detected
by GC‐MS as this compound can be sensitively observed through its
molecular ion at m/z 250 (55%) and its base peak at m/z 111 (van Beek,
unpublished). A somewhat similar adulteration scenario would be the
addition of patchoulol produced by transgenic Artemisia annua
plants.344 Should the price of PEO once again rise to $ 160/kg as has
happened in the past,13 such practices might occur.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
5 | O D O U R OF P A T C H O U L I O I L
5.1 | Constituents responsible for patchouli odour
Not less than 50 major patchouli‐themed perfumes were released
between 1917 and 2007.345 Much has been said and written about
the odour of patchouli essential oil and extracts,6,346 and regardless
of their origin, the qualitative olfactory properties of PEO are complex.
Arctander has published a pertinent olfactory assessment of patchouli
odour: “a dark orange or brownish‐coloured, viscous liquid, possessing
an extremely rich, sweet‐herbaceous, aromatic‐spicy and woody‐
balsamic odour. An almost wine‐like, ethereal‐floral sweetness in the
initial notes is characteristic of good oils, although this top note can
be absent or masked in freshly distilled, but otherwise good oils”.347
The odour is also described as "rich, heavy, sweet‐herbaceous,
aromatic/spicy/camphoraceous, woody, balsamic, root‐like and deli-
cately earthy",348 and “very intense, woody, sweet‐balsamic with
spicy and woody‐earthy undertones; displays extreme originality”.7
Odour evaluation of fractions from patchouli extracts or essential
oils may be biased by the presence of contaminants such as
anethole.349
Many authors have used a set of three basic odour descriptors
repeatedly: earthy, woody and camphoraceous.155,350 Whereas the
earthy, "cellar like" note is clearly reminiscent of mould, the woody
descriptor is more vague, since there are many natural products,
including fragrant wood extracts and synthetic odorants, which belong
to this category, all providing various nuances and effects (e.g., "dry
cedarwood", "warm agarwood", iso‐E Super, etc.). The "camphora-
ceous" attribute is sometimes found less desirable by perfumers, which
initiated the design of synthetic patchouli substitutes.351
The controversial contribution of (−)‐patchoulol to the odour of
PEO is undoubtedly a historical case. As early as 1875, this hitherto
unidentified compound was suspected to be odourless "if repeatedly
crystallised".16 This was confirmed in 1903.217 However one year
later it was stated in an inconclusive report of Schimmel & Co. that
patchoulol after repeated crystallisation still smelled of patchouli.
Nonetheless it was speculated that it might possibly be odourless
when pure.269 In 1949, a new investigation confirmed that recrystal-
lized patchoulol smelled characteristically of patchouli.148 Probably in
view of the earlier findings, the authors allowed for the possibility
that the odour was caused by minute amounts of degradation
products instead of patchoulol itself. The odourless character of
(−)‐patchoulol was again alleged by Roure Bertrand Dupont in
1973, possibly to support the decisive contribution of
norpatchoulenol 34, which had just been isolated and identified as

VAN BEEK AND JOULAIN
“the true carrier of the odour of patchouli oil”.193,232 However com-
petitors from the fragrance industry opposed this claim. Firmenich
stated that mainly patchoulol was responsible for the odour of
PEO with a minor role for norpatchoulenol. In a sniffing port analysis
the latter molecule was described as having an “intense, rooty,
352
cellar‐potato‐like odour”. Independently an academic group also
stated that patchoulol had a typical PEO odour and that
norpatchoulenol possessed a stronger odour but that it was not typ-
ical of the complete oil.186 Subsequently and possibly in response to
this paper, Teisseire weakened his own earlier claim and wrote that
pure patchoulol is “practically odourless”,353 a statement, which later
gained moderate support from colleagues at Givaudan: "patchoulol
... contributes very little to the olfactive character of this important
354
natural product". Meanwhile, chemists from IFF published their
extensive studies on PEO, which included a synthesis of racemic
patchoulol. This was found to have an odour “nearly identical” to that
of pure natural patchoulol.155 Further they confirmed the impor-
tance of norpatchoulenol, which possessed according to them a
“considerably more intense odour than that of patchoulol”. Addition-
ally they identified nortetracyclopatchoulol 35 (see § 3.3) as a new
155
powerful player in this fragrant orchestra. Some years before this
compound had already been patented by IFF.276 Its structure was
corrected about 40 years later by other authors who reisolated this
compound with the help of GC‐olfactometry (see § 3.3) and con-
firmed that it showed a “strong woody and earthy odour with warm
and sweet nuance”. 162
The one‐century old dispute concerning the odour of patchoulol
came finally to an end with the synthesis of both enantiomers of
patchoulol. This enabled to assess their odour unambiguously.355 The
synthetic, nature‐identical (−)‐(1R,3R,6S,7S,8S)‐patchoulol exhibited a
“strong, typical patchouli scent with an earthy, slightly camphoraceous,
powdery cellar note, which was practically indistinguishable from nat-
ural patchouli alcohol”. In contrast, the odour profile of the unnatural
(+)‐(1S,3S,6R,7R,8R)‐patchoulol was described to be “much weaker,
less characteristic, nearly indefinable and by no means reminiscent of
patchouli. It might however have a β‐santalol odour with a green
355
undertone”.
A somewhat rudimentary but nonetheless effective GC‐
olfactometric analysis of PEO allowed assigning intensities to approxi-
mately 15 GC‐eluted constituents. The scores ranged from odourless
(score 0), via weak (1) and well detectable (2) to intense (3). Patchoulol
356
was the only one rated 3. Thus the authors confirmed that
patchoulol is the main odour carrier, although they incorrectly
indicated it to be a racemate. It is noteworthy that the odour of the
major sesquiterpene hydrocarbons was described as well detectable
("gut wahrnehmbar"). Earlier Bruns had already carried out a more
extensive GC‐olfactometric analysis of PEO with 32 detected peaks
75
and their odour descriptions.
The detection thresholds measured for the three closely
related tertiary alcohols 33, 34 and 35 are important parameters
to consider when discussing their odour value and relative
contributions:52
(−)‐patchoulol 33: ~0.93 ng/L air
(+)‐norpatchoulenol 34: ~2.8 ng/L air
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
39
(+)‐nortetracyclopatchoulol 35: ~1.0 ng/L air
The authors stated that these figures are approximate and indeed
Mookherjee et al. arrived at a slightly different ranking.155 The concen-
trations given in the latter paper for 33, 34 and 35 of 30, 0.5 and
0.001% respectively are somewhat arbitrary and will vary considerably
in practice (see § 3.4). For 35 it seems too low as it can be easily
observed by GC‐MS in total PEO and can even reach a concentration
of ~1%, e.g., see Figure 7b.
Kraft pertinently reminded us that quite a number of people are
anosmic or have very different sensitivities for these natural
products.345 This is a rather frequent physiological phenomenon.
Secondly, and more importantly, in view of the many studies carried
out during the last three decades of the last century on the structure/
patchouli odour relationship of structurally related tertiary alcohols, the
probability that patchoulol is odourless is extremely low.155,351,357-360
Depending on the shape and size of pure patchoulol crystals,
sublimation eliciting odour perception may vary and could possibly
suggest that it is odourless. Together with anosmia, this hypothetical fea-
ture may account for the contradictory odour assessments of patchoulol
over almost 150 years, and should motivate further investigations.
In contrast with these "master contributors", very few odour
descriptions of other isolated constituents, minor or not, have been
published. Pogostol has been mentioned once as one of the major
contributors to the woody odour of PEO but this seems somewhat
doubtful.361 The "strong ambergris" odour of α‐patchoulone 99,
initially known as a hemi‐synthetic product,253 and later identified at
"trace" levels in PEO, and the "strong woody patchouli" odour carried
by β‐patchoulenone 39 (also as a "trace") have been mentioned.183,252
Not less interesting is the – still putative – contribution of
rotundone 40, an extremely potent compound with a detection
threshold of 8 ng/L in water, although 20% sensory panellists are
subject to anosmia.362,363 Actually, the contribution of 40 as a "wood,
peppery, agarwood‐like" compound to the odour performance of
PEO as a whole,364 has not been assessed yet. Cis‐ and trans‐
pentylcyclopropane carboxylic acids, respectively 61 and 62, are
potent odour contributors: when isolated, the cis‐isomer 61, which
occurs in the essential oil at ~200 ppm, is claimed to possess a
"patchouli‐like, animal, leathery note with good intensity and persis-
tence".183,271,279 The contribution of pogostone (syn. dhelwangin) 51
that occurs – like its minor analogs – in variable concentrations in
PEO,271,279 has not been reported so far, and is therefore supposed
to be negligible or nil.
Based on the structural similarity, one could expect the odour of
the major pyridine isomer 70 in PEO to be reminiscent of the "green,
earthy and carrot‐like" odour of epiguaipyridine, the major pyridine‐
sesquiterpene present in Cyperus scariosus essential oil (cypriol).251
However this is not borne out by the earlier odour description of 70
as being "weak pyridine‐like".268 More than one century ago,
Schimmel's chemists had detected a "base of stupefying odour".269
Patchouli pyridine 69 and guaipyridine 70 are described to possess
"off‐notes of nauseating, rooty‐quinolinic character".52 These pyridines
and other nitrogen‐containing compounds have therefore controversial
odour properties: some think they contribute to the aroma of patchouli
(Joulain, unpublished) while others disagree.183

40
Several sources have remarked that fresh PEO does not have
the best odour and that aged oils smell better.49,55,310,347,352 This
aspect has best been described by Guenther: ‘Freshly distilled oils
have a somewhat “green” and harsh note which, however, changes con-
siderably with time. Aging of the oil for a prolonged period will develop
that full, rich, and almost fruity note for which the best grades of patch-
ouly are renowned and so highly esteemed by expert perfumers’ as well
as ‘Aging of the oil is of utmost importance. A patchouly oil several
years old possesses a much finer and fuller odor than one freshly
27
distilled’. Freshly distilled essential oils often possess an off‐note
due the presence of highly volatile olfactorily‐undesirable com-
pounds like acetaldehyde, ammonia or hydrogen sulfide.18 These
preferentially evaporate the first days after distillation. However, a
continuous quality increase over a number of years is chemically dif-
ficult to envisage and we have never come across any clear explana-
tion or chemical and olfactory research to back up this long‐standing
observation. If PEO is properly packed (room temperature, dry, no
oxygen, no light, inert container), apart from some redox reactions,
dehydrations and subsequent hydrations, very little chemistry should
occur and the smell is therefore not expected to change noticeably.
However, there are some hints that the improvement occurs
primarily when the oil is not properly packed and dried. In that case
hydration, oxidation and partial polymerisation will occur and some
alteration and actual improvement of the odour might occur.
Howarth suggested that this is done on purpose: ‘The first rule when
it comes to odor is that patchouli gets better with age. This is key,
because as the odor matures it gets more round and loses some of
the harsher green notes. One can achieve this through a simple process
of aerating to start the oxidation process. Sometimes this “ageing” is
10
done at origin before sampling’.
An example is the formation of rotundone 40 by aerial oxidation of
guiaene 16.256,257 Thus the formation of traces of 40 might alter or
improve the overall aroma of PEO over time. In somewhat oxidised
PEOs, rotundone can occur in relatively high concentrations (~1%;
Figure 7c). A somewhat similar story could hold true for the strong
celery‐like smelling dihydrobovolide 95, which has been cited as a
contributor to the odour of PEO.155 Indeed, lactone 95 has a low
odour threshold of 480 pg/L in air and can be formed through oxida-
tion processes.365 However we have never been able to detect it in
any PEO or patchouli extract and the identification remains tentative.
More research is clearly desirable.
The same holds true for twelve compounds (Table 3B) reported
recently at trace levels in a commercial PEO of Indonesian origin,
using an undefined headspace sampling technique.165 Both
2‐methylisoborneol 79 and geosmin 80 with detection thresholds
(ppt levels) much lower than their actual concentrations are highly
potent elements and could contribute to the earthy/ musty odour.
However one should not rule out the possibility that 79 and 80 are
actually artefacts produced by cyanobacterial contamination of the
water used for the distillation, since tank storage of rainwater is a
common practice in PEO‐producing countries.
In the perfumery industry, differences of opinions are indeed not
scarce when expressing preferences, and patchouli is no exception. On
the one hand, some creative perfumers consider that the complexity
of a crude essential oil is the key to its attractiveness. In contrast, others
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
claim that removing acidic, phenolic and/or basic fractions, produces a
warm, herbacious, camphoraceous, woody‐patchouli odour that is
better than the odour of the original crude oil.252 Lowering the content
of certain sesquiterpene hydrocarbons carrying "a nasty top note" by
fractional distillation with concomitant enrichment of patchoulol and
norpatchoulenol is also claimed to provide a rectified oil having "no
green or earthy top note, but rather a mild sweet, woody, balsamic
and amber‐like odour".366 In summary, it appears that the typical
harsh‐musty character of crude PEO may or may not be appreciated
to the same extent by some perfumers in their creations.
5.2 | Clearwood™, a possible substitute for
patchouli oil?
The biotech company Amyris in Brazil in a partnership with
Firmenich produces Clearwood™, which according to a marketing
sheet is a “soft clean version of Patchouli without the earthy,
leathery and rubbery notes found in the natural oil”.164 It can be
used like patchouli or as a new woody building block at higher
dosages.164 The production is based on research carried out by
Firmenich over the past 15 years. Enzymes involved in the synthesis
of patchoulol and other sesquiterpenes were identified in P. cablin.
One cDNA was found to encode for a multifunctional patchoulol
synthase, which in combination with farnesyl pyrophosphate (FPP)
produced most of the sesquiterpenes found in PEO in similar yields
and ratios.367 Part of the underlying research was patented.368,369
Through a recombinant DNA approach, the required enzymes are
brought to expression in a transformed host organism, in this case
a yeast (Saccharomyces cerevisiae).370 The yeast should also be
capable of producing the starting material FPP and is grown on a
medium based on cane sugar. This approach is now being used for
many more valuable plant products.371 A new CAS RN [1450625‐
49‐6] was created for this product “Oils, Patchouli, patchoulol
synthase‐modified Saccharomyces cerevisiae‐fermented, from
carbohydrates” but rather surprisingly in the marketing sheet the
CAS RN [84238‐39‐1] is used, which refers to P. cablin extracts
(see Introduction). However, Clearwood™ is prepared via a GMO
process and is certainly not a P. cablin extract.
Chemically Clearwood™ consists of the regular sesquiterpene
hydrocarbons found in PEO plus pogostol and patchoulol. It contains
neither the acids, bases and pogostone present in PEO nor the
olfactorally important norpatchoulenol and nortetracyclopatchoulol
(see § 5.1), which probably apart from pogostone, all contribute to
the “dirty” odour of PEO. In contrast, it contains several oxygenated
sesquiterpenes, including patchoulol ethyl ether, which are not present
in regular PEO (van Beek, unpublished). As a result, the natural status
of Clearwood™ would certainly be challenged if one considers CAS
RN [84238‐39‐1]. Clearwood™ has a different “cleaner” odour and
the statement that it can be used as a new woody building block or
“PEO extender” is probably closer to the truth than that it can be used
like patchouli. Indeed, a recent patent claims the use of compounded
mixtures containing inter alia highly potent trace elements newly found
in PEO, with the aim of restoring somehow Clearwood™'s lack of
richness.165

VAN BEEK AND JOULAIN
6 | TOXICOLOGY AND SKIN SENSITISATION
The skin contact allergy of essential oils or their constituents is a concern
in fragrance compounding, and PEO is not an exception.309,372,373 Neat
or as the main constituent of PEO, patchoulol is a fragrance ingredient
used in decorative cosmetics, fine fragrances, shampoos, toilet soaps
and other toiletries as well as in non‐cosmetic products such as house-
hold cleaners and detergents. In neat form, its use annual worldwide
use is 0.5‐1 metric tonne, whereas the consumption of the essential oil
is much bigger at ~1300 tonnes. Belsito et al. have made an assessment
of the general toxicological and dermatological risk of many terpene
alcohols including patchoulol‐containing products.374 Their conclusion
was that patchoulol has a low order of acute toxicity with dermal and
oral NOAELs (no observed adverse effect levels) of 50 mg/kg body
weight/day or greater and is negative in mutagenicity and genotoxicity
tests. At current use levels, patchoulol is non‐irritating and sensitisation
potential is generally low. In a separate note the results specific to
patchoulol were summarised by Bhatia et al.375 In a rat study, PEO orally
administered for 90 days at a level in excess of at least 100 times the
maximum estimated daily dietary intake in man evoked no adverse
effect on growth, food consumption, haematology, blood chemistry,
liver and kidney weights or on gross and microscopic appearance of
major organs at autopsy.376
Over time, various groups have investigated PEO in more detail
with respect to the risk of triggering allergic reactions on the skin.
Nakayama et al. found 3 “strongly positive” and 8 “weakly positive”
reactions to “Patchouli oil” (unknown test concentration) in patch tests
with 183 patients.377 The number of patients, mostly Japanese
women, with pigmented cosmetic dermatitis thought to be caused in
part by PEO, decreased significantly after 1978 when major cosmetic
manufacturers began to eliminate strong contacts sensitizers (e.g.,
cinnamaldehyde) from their products.378 Clinical data published in
2002 reported that 0.8% positive reactions to PEO (10% in petrolatum)
in 1606 consecutive patients were observed.379 A more recent study
on patch results with PEO identified 0.6% positive reactions in 2446
consecutively tested patients and 1.4% positive reactions in 828
patients tested in the context of a special series.380 For assessing skin
contact allergy of cosmetics, two "fragrances mixes" (14 components
in total), the 26 allergens,381 which need to declared in cosmetics,
and several essential oils were tested. When PEO was patch‐tested
in this study (10% concentration in petrolatum), 53 patients out of
382
5539 (1%) gave a positive reaction. It is regrettable that, no informa-
tion was given in this report about the intensities of the reactions, and
none of these reports gave any information about the quality of the
tested essential oils (origin, genuineness, age, peroxide value, etc.).
Some of the above studies have been summarised by the Scientific
Committee on Consumer Safety of the EU.383
Among the constituents ever reported in PEO, at least six are
restricted in fragrance compounding: limonene, linalool, citronellol,
farnesol,206 eugenol,271 and isophorone.252 Whereas limonene, linal-
ool, citronellol, farnesol and eugenol are suspected skin sensitisers
384
with restricted use levels or purity requirements with regard to
the level of oxidation, isophorone 91 is officially classified as a CMR
(carcinogen, mutagen, reprotoxic) and should not be used at all as a
fragrance ingredient.385 However eugenol 68 occurs only at low ppm
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
41
levels in genuine PEO (see § 3.3) and the presence of isophorone and
farnesol has not been confirmed to date. The other above cited
compounds are trace elements in patchouli (<150 ppm) and can be
detected and quantified accurately only by using a validated specific
GC‐MS method.386 One should not rule out however that the
presence of these compounds in PEO is the result of contamination
during harvesting, distillation or manufacturing.
The European Chemicals Agency (ECHA) is the driving force
among regulatory authorities in implementing the European Union's
groundbreaking chemicals legislation for the benefit of human health
and the environment as well as for innovation and competitiveness.
Just as for any other chemical entering the EU or used in the EU, ECHA
provides information on PEO and addresses toxicological concerns.387
7 | CO NC LUSIO N
Among the sesquiterpene‐rich essential oils in the category "natural
woody commodities", patchouli is of major importance for the
fragrance industry. Similar to other important raw materials of this type
such as vetiver and sandalwood oils (and to a lesser extent agarwood),
sesquiterpenes represent more than 95% of PEO, including at least six
nor‐ and two seco‐sesquiterpenes, but its composition has not yet been
investigated in‐depth to the same level. Hence, whereas more than 150
volatile constituents have been identified in vetiver,5 sandalwood,3 and
agarwood,4 including many minor or even trace elements of high olfac-
tory relevance, the number of such compounds identified so far in PEO
is unfortunately smaller. Considering that the main vectors of the
earthy‐mouldy note, namely the trio patchoulol 33, norpatchoulenol
34 and nortetracyclopatchoulol 35, alone cannot account for the rich-
ness and high complexity of PEO odour, there is an on‐going need to
identify more high impact odour‐donating components of PEO, includ-
ing carbonyl‐containing sesquiterpenes such as rotundone 40.
It is noteworthy that only six important odour contributors have been
conclusively identified during a 30‐year period (1962‐92) represented by
~25% of the references cited in the present review, and no such compo-
nents – with exception of rotundone – have been identified during the last
25‐year period representing ~65% of the references! Notwithstanding the
very recent findings concerning ultra‐trace constituents,165 this is a para-
dox, when one considers the availability of efficient GC‐olfactometry
devices to pinpoint key odorants, and improved separation techniques as
well as more sensitive and powerful spectroscopic tools for the identifica-
tion of complex chemical structures. The launching in the late 1980s of low
cost GC‐MS systems, leading to a proliferation of lower quality publica-
tions in the domain of essential oil analysis, partially accounts for this
regrettable situation. More GC‐MS analyses will by themselves not provide
the structures of the many still unknown PEO constituents, even if occur-
ring at the 1‐2% level.
Furthermore, what happens in fresh patchouli leaves during the
"drying‐fermentation" step and subsequent storage prior to steam
distillation in relation to PEO odour is still enigmatic and remains to
be clarified, despite recent reports.388-390 This holds also true for the
ill‐understood increase in olfactory quality of PEO after years of
storage. Finally, the fate of some genuine leaf constituents – already
identified or pending identification – during the distillation process also
needs to be better understood. The present review is therefore a

42
strong invitation for skilled phytochemists to reopen, in collaboration
with other scientists, the patchouli case.
8 | SUMMARY OF SUPPLEMENTARY
INFORMATION
The Supplementary Information belonging to this paper consists of:
Table S1. List of non‐cited patchouli‐related papers of which the
focus is outside the scope of this review or contain possibly erroneous
analytical chemical data. Papers are presented in chronological order.
Figure S1. Complete chromatogram of PEO B (see also Figure 7b),
i.e., including monoterpenes.
Figure S2. Chromatogram of oil B (Figure 7b) on a dimethyl
polysiloxane phase (DB1).
Figure S3. Chromatogram of oil B (Figure 7b) on a polyethylene
glycol phase (Wax).
Figure S4. Complete gas chromatogram of a commercial steam
distilled PEO on a 30 m DB5 stationary phase with a different temper-
ature programme.
Figure S5. Chromatogram of heavily adulterated commercial PEO
on a DB5 phase.
ACKNOWLEDGEMEN TS
We gratefully acknowledge Bo Chen, Xiao Wei, Bruno Maurer, Charles
Cornwell, Arnaldo Bandoni, Philip Kraft, Robin Clery, Shashikumar
Paknikar, Brian Lawrence, Toshio Hasegawa, Anil Pant, Arjen Schots,
David Leach, Suaib Luqman, Andre Krause, Annette Bierman, Pat
Sandra, Luigi Mondello, Lydia Ziegler, Sascha Beutel, Laura McGregor,
Eranki Prakasa Rao, Louis Peyron, Sabrina Boutefnouchet, Keita Tanaka
and Liang Xiong for providing information, literature or translations.
Without their generous help this review would not be the same. We
are greatly indebted to Maarten Posthumus who kindly provided us with
the results of many of his unpublished but very high‐quality analyses of
patchouli oils and the corresponding chromatograms some of which
made it into this review (Figure 7a‐c). Last but not least we wish to thank
the late Hans Siwon who set up his own patchouli distillation facility in
Indonesia and carried through several innovations leading to oils with dif-
ferent olfactory properties. Over a period of ten years, he gave both
authors many high‐quality genuine patchouli oils and proverbially speak-
ing he planted the small seed that eventually led to this review.
ORCID
Teris A. van Beek http://orcid.org/0000-0002-9843-7096
Daniel Joulain http://orcid.org/0000-0001-9662-9732
RE FE R ENC E S
1. Joulain D, Tabacchi R. Lichen extracts as raw materials in perfumery.
Part 1: oakmoss. Flavour Frag J. 2009;24:49‐61.
2. Joulain D, Tabacchi R. Lichen extracts as raw materials in perfumery.
Part 2: treemoss. Flavour Frag J. 2009;24:105‐116.
3. Baldovini N, Delasalle C, Joulain D. Phytochemistry of the heartwood
from fragrant Santalum species: a review. Flavour Frag J. 2011;26:7‐26.
4. Naef R. The volatile and semi‐volatile constituents of agarwood, the
infected heartwood of Aquilaria species: A review. Flavour Frag J.
2011;26:73‐89.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
5. Belhassen E, Filippi J‐J, Brévard H, Joulain D, Baldovini N. Volatile
constituents of vetiver: a review. Flavour Frag J. 2015;30:26‐82.
6. Anonis DP. Woody notes in perfumery: patchouli in fragrances, part II.
Perfum Flavor. 2007;32:30‐35.
7. Haarmann & Reimer. Guide to Fragrance Ingredients. Vol. 4. Hamburg:
Gloess Publishing; 1985;113.
8. Lawrence BM. A preliminary report on the world production of some
selected essential oils and countries. Perfum Flavor. 2009;34:38‐44.
9. Anonymous. Socio‐economic committed report on patchouli ‐
Pogostemon cablin (Blanco) Benth. IFEAT World ‐ International Feder-
ation of Essential Oils & Aroma Trades, May edition. 2014;4‐5.
10. Howarth J. Natural product supply bulletin: patchouli, Mexican lime
and the Crimean conflict. Perfum Flavor. 2015;40‐January:32‐37.
11. Lawrence BM. Personal Communication 2016.
12. Swamy MK, Mohanty SK, Sinniah UR, Maniyam A. Evaluation of
patchouli (Pogostemon cablin Benth.) cultivars for growth, yield and
quality parameters. J Essent Oil‐Bear Plants. 2015;18:826‐832.
13. Frister T, Beutel S. Moschusduft und Patchouliöl ‐ Zwei Duftbausteine
fur Parfümerie und Kosmetik. Chem unserer Zeit. 2015;49:2‐9.
14. Verlet N. Commercialization of essential oils and aroma chemicals. In
A Manual on the Essential Oil Industry, Tuley de Silva K (ed.). United
Nations Industrial Development Organization: Vienna, 1995;203‐232.
15. Rochleder F. Phytochemie. Leipzig: Verlag von Wilhelm Engelmann;
1854;155.
16. Hunt R, Rudler FW. Ure's Dictionary of Arts, Manufactures, and Mines.
III. 7th edition. London: Longmans, Green, and Co.; 1875;516.
17. Anonymous. Patchouli (Pogostemon Patchouli, var. suavis). Bull Misc Inf
(Royal Bot Gardens, Kew). 1889;135‐139.
18. Gildemeister E, Hoffmann F. Die Aetherischen Oele. Berlin: Verlag
Julius Springer; 1899;855‐859.
19. Beilstein F. Handbuch der Organischen Chemie. Dritte Auflage, dritter
Band. Hamburg und Leipzig: Verlag von Leopold Voss; 1897;514 & 538.
20. Anonymous. U.S. Food and Drug Administration, Code of Federal
Regulations, Title 21, Vol. 3, Part 172 ‐ Food additives permitted for
direct addition to food for human consumption. Sec. 172.510 Natural
flavoring substances and natural substances used in conjunction with
flavors (21CFR172.510). 2016.
21. Hall RL, Oser BL. Recent progress in the consideration of flavoring
ingredients under the food additives amendment. III. GRAS sub-
stances. Food Technol. 1965;19:151‐197.
22. Anonymous. Patchouli oil, https://echa.europa.eu/fr/substance‐infor-
mation/‐/substanceinfo/100.109.383, accessed on April 29, 2017.
23. Anonymous. Inventory of Existing Cosmetic Ingredients in China
(CFDA, Decree No. 105, 2015) ‐ 02634 Pogostemon cablin oil,
http://www.cirs‐reach.com/news‐and‐articles/new‐inventory‐of‐
existing‐cosmetic‐ingredients‐in‐china‐launched‐(iecic‐2015,‐final‐ver-
sion).html, accessed on May 9, 2017.
24. Zörnig H. Arzneidrogen. Vol. II. Leipzig: Werner Klinkhardt;
1911;203‐206.
25. Parry EJ. The Chemistry of Essential Oils and Artificial Perfumes. Vol. I.
Fourth Edition. London: Scott, Greenwood and Son; 1921;250‐255.
26. Gildemeister E, Hoffmann F. Die Ätherischen Öle. Dritter Band.
Dritte Auflage. Miltitz bei Leipzig: Verlag der Schimmel & Co.
Aktiengesellschaft; 1931;893‐908.
27. Guenther E. The Essential Oils. Vol. III. Princeton: Van Nostrand Co.
Inc.; 1949;552‐575.
28. Soepadyo R, Tan HT. Patchouli, a profitable catch crop. World Crops.
1968;3:48‐54.
29. Kumar A, Gauniyal AK, Virmani OP. Cultivation of Pogostemon cablin
for its oil. Curr Res Med Aromat Plants. 1986;8:79‐86.
30. Husain A, Virmani OP, Sharma A, Kumar A, Misra LN. Major Essential
Oil‐bearing Plants of India. Lucknow: Central Institute of Medicinal
and Aromatic Plants; 1988, Chapter 35 ‐ Patchouli Oil, 187‐191.

VAN BEEK AND JOULAIN
31. Husain A. Essential Oil Plants and their Cultivation. Lucknow: Central
Institute of Medicinal and Aromatic Plants; 1994;136‐140.
32. Maiti S, Raju S, Greta KA, Mandal K. Good Agricultural Practices for
Patchouli, Geranium and Lemongrass. Boriavi, Anand ‐ 387310, Gujarat,
India. 2006;1‐7.
33. Robbins SRJ. Selected markets for the essential oils of patchouli and
vetiver. Report of the Tropical Products Institute, G167, London, 1982.
34. Manheimer S. Long live Patchouli! Cosmet World News. 1983;33.
35. Akhila A, Tewari R. Chemistry of patchouli oil: a review. Curr Res Med
Aromat Plants. 1984;6:38‐54.
36. Boelens MH. Chemical and sensory evaluation of trace compounds in
naturals, In Proceedings of 13th International Congress of Flavours, Fra-
grances and Essential Oils, Istanbul, Turkey, Boelens Aroma Chemical
Information Service (BACIS), 1995;177‐185.
37. Lawrence BM. Progress in essential oils ‐ patchouli oil. Perfum Flavor.
1976;1:20.
38. Lawrence BM. Recent progress in essential oils. Perfum Flavor.
1978;2:44‐50.
39. Lawrence BM. Progress in essential oils. Perfum Flavor.
1981;6:73‐74,76.
40. Lawrence BM. Progress in essential oils. Perfum Flavor. 1990;15:
(March/April) 75‐77.
41. Lawrence BM. Progress in essential oils. Perfum Flavor.
1995;20(3):67‐68,70,72.
42. Lawrence BM. Progress in essential oils. Perfum Flavor.
2002;27(3):48,50‐54,56‐60,62,64‐66,68‐69.
43. Lawrence BM. Progress in essential oils ‐ Patchouli oil and extracts.
Perfum Flavor. 2014;39:46,48,50‐52.
44. Bhat KKS. Agrotechnology of aromatic plants. In A Manual on the Essential
Oil Industry, Tuley de Silva K (ed.). UNIDO: Vienna, 1995;31‐32.
45. Leung AY, Foster S. Encyclopedia of Common Natural Ingredients
Used in Food, Drugs, and Cosmetics. New York: Wiley & Sons;
1996;411‐413.
46. Das K. Patchouli (Pogostemon Cablin Benth) Oils. In Essential Oils in
Food Preservation, Flavor and Safety, Preedy V (ed.). Academic Press:
Amsterdam, 2016;633‐639.
47. Weiss EA. Essential Oil Crops. Wallingford: CAB International; 1997,
138‐154.
48. Oyen LPA. Pogostemon Desf. In Essential‐Oil Plants, Oyen LPA, Dung
NX (ed.). Backhuys Publishers: Leiden, 1999;151‐157.
49. Panda H. Patchouli. New Delhi: Asia Pacific Business Press Inc.;
2005;85‐93.
50. Wu Y‐g, Guo Q‐s, Zheng H‐q. Textual research on history of introduc-
tion and herbal medicine of Pogostemon cablin. China J Chin Materia
Med. 2007;32:2114‐2117.
51. Murugan R, Livingstone C. Origin of the name ‘patchouli’ and its his-
tory. Curr Sci. 2010;99:1274‐1276.
52. Ohloff G, Pickenhagen W, Kraft P. Scent and Chemistry ‐ The Molecular
World of Odors. Weinheim: Wiley‐VCH; 2012;251‐258.
53. Chakrapani P, Venkatesh K, Chandra SSB, et al. Phytochemical, pharma-
cological importance of Patchouli (Pogostemon cablin (Blanco) Benth)
an aromatic medicinal plant. Int J Pharm Sci Rev Res. 2013;21:7‐15.
54. Swamy MK, Sinniah UR. A comprehensive review on the phytochem-
ical constituents and pharmacological activities of Pogostemon cablin
Benth.: an aromatic medicinal plant of industrial importance.
Molecules. 2015;20:8521‐8547.
55. Ramya HG, Lpalanimuthu V, Singla R. An introduction to patchouli
(Pogostemon cablin Benth.) ‐ a medicinal and aromatic plant: it's impor-
tance to mankind. Agric Eng Int. 2013;15:243‐250.
56. Anonymous. Patchouli (Pogostemon cablin Benth.) Family‐Lamiaceae,
Assam Small Farmers' Agri Business Consortium, http://
assamagribusiness.nic.in/NEDFi/map6.pdf, accessed on September
28, 2016.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
43
57. Swamy MK, Sinniah UR. Patchouli (Pogostemon cablin Benth.):
botany, agrotechnology and biotechnological aspects. Ind Crops Prod.
2016;87:161‐176.
58. Chen Y, Wu Y‐G, Xu Y, et al. Dynamic accumulation of sesquiterpenes
in essential oil of Pogostemon cablin. Rev Bras Farmacogn.
2014;24:626‐634.
59. Tripathi YC, Hazarika P. Impact of harvesting cycle, maturity stage,
drying and storage on essential oil content of patchouli leaves
grown in Northeast region of India. J Essent Oil‐Bear Plants.
2014;17:1389‐1396.
60. Hussin N, Mondello L, Costa R, Dugo P, Yusoff NIN, Yarmo MA,
Wahab AA, Said M. Quantitative and physical evaluation of patchouli
essential oils obtained from different sources of Pogostemon cablin.
Nat Prod Commun. 2012;7:927‐930.
61. (a) Blank AF, Sant'ana TCP, Santos PS, et al. Chemical characterization
of the essential oil from patchouli accessions harvested over four
seasons. Ind Crops Prod. 2011;34:831‐837; (b) de Sant'ana TCP, Blank
AF, Vieira SD, Arrigoni‐Blank MF, de Jesus HCR, Alves PB. Influência
do armazenamento de folhas secas no óleo essencial de patchouli.
Quim Nova. 2010;33:1263‐1265.
62. Zhang L‐G, Zhang C, Ni L‐J, Yang Y‐J, Wang C‐M. Rectification extrac-
tion of Chinese herbs' volatile oils and comparison with conventional
steam distillation. Sep Purif Technol. 2011;77:261‐268.
63. Santos AV, Arrigoni‐Blank MF, Blank AF, et al. Mass multiplication of
Pogostemon cablin (Blanco) Benth genotypes and increase of essential
oil and patchoulol yield. Ind Crops Prod. 2010;32:445‐449.
64. Paul A, Thapa G, Basu A, Mazumdar P, Kalita MC, Sahoo L. Rapid plant
regeneration, analysis of genetic fidelity and essential aromatic oil
content of micropropagated plants of Patchouli, Pogostemon cablin
(Blanco) Benth. ‐ An industrially important aromatic plant. Ind Crops
Prod. 2010;32:366‐374.
65. Kumar A, Srivastava NK, Kalra A, Mishra BN, Luqman S, Khanuja SPS.
Allocation of 14CO2 into primary metabolites and leaf terpenoid in
relation with storage duration in patchouli (Pogostemon cablin). J Plant
Biol (New Delhi, India). 2009;36:33‐40.
66. Kumar A. Effect of soaking time of Patchouli leaves on oil yield and
chemical composition. Indian Perfum. 2009;53:27‐28.
67. Rana VS, Blazquez MA. Volatile leaf essential oil of commercial
Pogostemon cablin Benth. cultivated in Manipur. Indian Perfum.
2009;53:30‐32.
68. Kongkathip N, Sam‐ang P, Kongkathip B, Pankaew Y, Tanasombat M,
Udomkusonsri P. Development of patchouli extraction with quality
control and isolation of active compounds with antibacterial activity.
Kasetsart J (Nat Sci). 2009;43:519‐525.
69. Rekha K, Bhan MK, Dhar AK. Development of erect plant mutant with
improved patchouli alcohol in patchouli [Pogostemon cablin (Blanco)
Benth]. J Essent Oil Res. 2009;21:135‐137.
70. Donelian A, Carlson LHC, Lopes TJ, Machado RAF. Comparison of
extraction of patchouli (Pogostemon cablin) essential oil with supercrit-
ical CO2 and by steam distillation. J Supercrit Fluids. 2009;48:15‐20.
71. Ramachandra KM, Farooqi AA, Srinivasappa KN. Effect of gamma rays
on growth, yield and oil composition in patchouli (Pogostemon patch-
ouli Pellet.). Indian Perfum. 2008;52:58‐60.
72. Prakash O, Joshi S, Shukla AK, Pant AK. Sesquiterpenoid rich essential
oil from the leaves of Pogostemon patchouli pellet grown organically
under tarai conditions. J Essent Oil‐Bear Plants. 2007;10:157‐161.
73. Bhaskar S, Kumar TV. Agronomic bottlenecks, genetic barriers and
marketing impediments in patchouli Pogostemon patchouli production.
J Med Aromat Plant Sci. 2000;22:396‐403.
74. Sugimura Y, Ichikawa Y, Otsuji K, et al. Cultivarietal comparison of
patchouli plants in relation to essential oil production and quality.
Flavour Frag J. 1990;5:109‐114.
75. Bruns K. Contribution to the study and quality evaluation of patchouli
oil. Parfüm Kosmet. 1978;59:109‐115.

44
76. Singh R, Singh M, Srinivas A, Rao EVSP, Puttanna K. Assessment of
organic and inorganic fertilizers for growth, yield and essential oil
quality of industrially important plant patchouli (Pogostemon cablin)
(Blanco) Benth. J Essent Oil‐Bear Plants. 2015;18:1‐10.
77. von Rechenberg C. Theorie der Gewinnung und Trennung der
Ätherischen Öle durch Destillation. Miltitz bei Leipzig: Selbstverlag
von Schimmel & Co.; 1910;441.
78. Ambrose DCP, Annamalai SJK, Naik R. Effect of drying on the volatile
oil yield of patchouli. Indian J Sci Technol. 2013;6:5559‐5562.
79. Rulianah S, Meilany D, Maryanty Y, Purwanto RE, Nanda D, Silvia W.
Production of pachouli oil by fermentation methode using
Phanerochaete chrysosporium with kieserite as sustitution MgSO4. Int
J Eng Res Developm. 2015;11:14‐19.
80. Reliance Life Sciences PVT LTD, Process for increased patchulol con-
tent in essential oil of Pogostemon cablin, World Patent 2007/080606,
January 16, 2006.
81. Ravindra NS, Ramesh SI, Gupta MK, et al. Evaluation of somaclonal
variation for genetic improvement of patchouli (Pogostemon patchouli),
an exclusively vegetatively propagated aromatic plant. J Crop Sci
Biotech. 2012;15:33‐39.
82. Misra M. Regeneration of patchouli (Pogostemon cablin Benth.) plants
from leaf and node callus, and evaluation after growth in the field.
Plant Cell Reports. 1996;15:991‐994.
83. Sarma A, Sarma TC. Patchouli oil recovery and effect of leaf ageing.
Indian Perfum. 2003;47:151‐154.
84. Misra M. Growth, photosynthetic pigment content and oil yield of
Pogostemon cablin grown under sun and shade conditions. Biologia
Plantarum. 1995;37:219‐223.
85. Misra M. Application of gibberellin to Pogostemon cablin plants:
growth, photosynthetic pigment content and oil yield. Biologia
Plantarum. 1995;37:635‐639.
86. Hussain AI, Anwar F, Nigam PS, et al. Antibacterial activity of some
Lamiaceae essential oils using resazurin as an indicator of cell growth.
LWT ‐ Food Sci Technol. 2011;44:1199‐1206.
87. Manjunatha R, Farooqi AA, Vasundhara M, Srinivasappa KN. Effect of
biofertilizers on growth, yield and essential oil content in patchouli
(Pogostemon cablin Pellet.). Indian Perfum. 2002;46:97‐104.
88. Singh M, Ganesha Rao RS. Influence of sources and doses of N and K
on herbage, oil yield and nutrient uptake of patchouli [Pogostemon
cablin (Blanco) Benth.] in semi‐arid tropics. Ind Crops Prod.
2009;29:229‐234.
89. Yang Y, Kong W, Feng H, et al. Quantitative and fingerprinting analy-
sis of Pogostemon cablin based on GC‐FID combined with
chemometrics. J Pharm Biomed Anal. 2016;121:84‐90.
90. Benveniste B. Indonesian oil of patchouli. Perfum Flavor. 1980;5:43‐45.
91. Muyassaroh, Daryono ED, Hudha MI. Determinating patchouli alcohol
of patchouli oil using distillation technique. Int J ChemTech Res.
2016;9:108‐116.
92. Singh M. Effect of potassium on growth and yield of patchouli
[Pogostemon cablin (Blanco) Benth.]. J Spices Aromatic Crops.
2014;23:76‐79.
93. Saha P, Chaudhuri A, Yadav GS, Babu S. Effect of nitrogen levels and
planting geometry on growth, herbage and oil yields of patchouli
(Pogostemon cablin Benth.) under humid sub‐tropical climate of North
Eastern Himalayas. Ann Agric Res New Series. 2014;35:274‐279.
94. Asangi H, Vasundhara M. Effect of pinching and growth regulators on
growth, herbage yield, essential oil content and oil yield of patchouli
(Pogostemon patchouli Pellet.). J Hortl Sci. 2013;8:214‐216.
95. Bhaskar S. Growth, herbage and oil yield of patchouli (Pogostemon
patchouli) as influenced by cultivars and nitrogen fertilization. Indian
Perfum. 1995;39:35‐38.
96. Saha BN, Dutta SC, Bordoloi DN, Mathur RK. Introduction of patch-
ouli in Itanagar. Indian Perfum. 1989;33:14‐19.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
97. Saha BN, Dutta SC, Bordoloi DN, Mathur RK. Prospect of growing
patchouli (Pogostemon patchouli) in Arunachal Pradesh ‐ Effect of
nitrogen on its yield. Indian Perfum. 1992;36:57‐60.
98. Hamidi N. Extraction of essential oils from patchouli plant using
advanced techniques of microwave‐assisted hydrodistillation. J Engin
Appl Sci. 2016;11:796‐799.
99. (a) Singh M, Rao RSG, Ramesh S. Influence of nitrogen and nitrogen
carrier on herbage, oil yield and nitrogen uptake of patchouli
[Pogostemon cablin (Blanco) Benth.] in semi‐arid tropical condition.
J Spices Aromatic Crops. 2007;16:106‐110. (b) Rao EVSP, Rao RSG,
Narayana MR, Ramesh S. Influence of shade on yield and quality of
patchouli (Pogostemon cablin) oil. Indian Perfum. 1997;41:164‐166.
100. Singh M. Effect of organic and inorganic fertilizers on growth, yield
and nutrient uptake of patchouli [Pogostemon cablin (Blanco) Benth.]
in a semi‐arid tropical climate. J Spices Aromatic Crops. 2011;20:48‐51.
101. Singh M, Guleria N. Effect of cultivars and fertilizer levels on growth,
yield and quality of patchouli [Pogostemon cablin (Blanco) Benth.]
under shaded condition. J Spices Aromatic Crops. 2012;21:174‐177.
102. Singh M. Effects of the amount of water applied, method of irrigation,
and rate of N‐application, on herbage and oil yields, N‐uptake, and
water‐use efficiency in patchouli [Pogostemon cablin (Blanco) Benth.]
grown under semi‐arid tropical conditions. J Hortl Sci Technol.
2012;87:441‐444.
103. Bhaskar S, Vasantha Kumar T, Srivastava HC. Effect of growth regula-
tors on production of herbage and oil in patchouli (Pogostemon
patchouli). Indian Perfum. 1997;41:98‐101.
104. Arpana J, Bagyaraj J, Prakasa Rao EVS, Parameswaran TN, Abdul
Rahiman B. Symbiotic response of patchouli (Pogostemon cablin
(Blanco) Benth.) to different arbuscular mycorrhizal fungi. Adv
Environm Biol. 2008;2:20‐24.
105. Ramachandra KM, Vasundhara M, Farooqi AA, Srinivasappa KN. Eval-
uation of patchouli (Pogostemon patchouli Benth.) varieties in relation
to different plant densities. Indian Perfum. 2002;46:7‐14.
106. Arpana J, Bagyaraj DJ, Prakasa Rao EVS, Parameswaran TN, Abdul
Rahiman B. Response of patchouli to inoculation with an arbuscular
mycorrhizal fungus and plant growth promoting rhizomicroorganisms.
J Soil Biol Ecol. 2014;34:70‐83.
107. Schrader T, Murasira P, Jouitteau F. Rwanda Agribusiness case: Patch-
ouli‐Biolandes. Production d'huile essentielle de patchouli dans le
nord du Rwanda. 2012, 23 pages, http://library.wur.nl/WebQuery/
wurpubs/fulltext/264607.
108. Costa GA, Carvalho Filho JLS, Deschamps C. Rendimento e composição
do óleo essencial de patchouli (Pogostemon cablin) conforme o tempo
de extração. Rev Bras Pl Med, Campinas. 2013;15:319‐324.
109. Chacko M. Characterization, evaluation and micropropagation of
Patchuli (Pogostemon patchouli Benth.): An important aromatic plant
in South India. Ph.D. Thesis, Mahatma Gandhi University, Kottayam,
Kerala, India, 2009.
110. Singh M. Influence of spacing and intercropping on biomass and
essential oil yield of patchouli [Pogostemon cablin (Blanco) Benth.].
J Spices Aromatic Crops. 2008;17:235‐239.
111. Magalhaes MT, Mendes PH, Wilberg VC. O óleo essencial de patchuli
da Amazônia. Acta Amazonica. 1976;6:467‐469.
112. Masetto MAM, Deschamps C, Castro LWP, Monteiro R, Guerios IT,
Sheer AdP, Côcco LC, Yamaemoto C. Rendimento e qualidade do óleo
essencial de Pogostemon cablin Benth em diferentes períodos de
secagem, In Proceedings of IV Simpósio Brasileiro de Óleos Essenciais,
Fortaleza, 2007.
113. (a) Nuryani Y. Characteristic of four accessions of patchouli
(Pogostemon cablin). Indon Agric Res Abstr. 2010;27:11‐12. (b) Rahman
M, Shoeb M, Nandi NC, Alamgir M, Husain M. Introduction of patch-
ouli, Pogostemon cablin (Blanco) Benth : an essential oil bearing plant
in Bangladesh. Bangladesh J Sci Ind Res. 2001;36:14‐18.
114. Wu Y, Li M. Induction of tetraploid plants of Pogostemon cablin
(Blanco) and its quality evaluation. Phcog J. 2013;5:281‐285.

VAN BEEK AND JOULAIN
115. Maheshwari ML, Vasantha Kumar T, Sharma N, Chandel KPS. Patch-
ouli ‐ an Indian perspective. Indian Perfum. 1993;37:9‐11.
116. Doraswamy K. Curing and distillation of patchouli. Indian Oil Soap J.
1967;32:211‐216.
117. Anilkumar M. Influence of growth regulators on growth and oil con-
tent of patchouli (Pogostemon cablin Benth.). M.Sc. Thesis, Univ. of
Agric. Sci., Dharwad, India, 2005.
118. Karimi A. Characterization and antimicrobial activity of patchouli
essential oil extracted from Pogostemon cablin [Blanco] Benth.
(lamiaceae). Adv Environm Biol. 2014;8:2301‐2309.
119. Zaman M, Ahmed M, Gogoi P. Effect of Bokashi on plant growth, yield
and essential oil quantity and quality in Patchouli (Pogostemon cablin
Benth.). Biosciences, Biotechnol Res Asia.
2010;7:383‐387.
120. Thai TH, Moi LD, Hoi TM, Muselli A, Casanova J. Production et com-
position de l'huile essentielle de Pogostemon cablin en relation avec la
récolte. Revue Pharmaceutique. 1999;31‐35.
121. Karim MF, Banerjee S, Poddar MK. Does patchouli oil change blood
platelet monoamine oxidase‐A activity of adult mammals? J Physiol
Sci. 2017; ahead of print: doi: 10.1007/s12576‐12017‐10534‐z.
122. Mahanta JJ, Chutia M, Sarma TC. Study on weed flora and their
influence on patchouli (Pogostemon cablin Benth.) oil and patchoulol.
J Plant Sci. 2007;2:96‐101.
123. Verma RS, Padalia RC, Chauhan A. Assessment of similarities and
dissimilarities in the essential oils of patchouli and Indian Valerian.
J Essent Oil Res. 2012;24:487‐491.
124. Sarma TC. Patchouli ‐ an aromatic oil crop for development of
perfumery industry in Northeast India. In Aromatic Plants from Asia ‐ their
Chemistry and Application in Food and Therapy, Jirovetz L, Dung NX,
Varshey VK (ed.). Har Krishan Bhalla & Sons: Dehradun, 2007;187‐201.
125. Puttanna K, Prakasa Rao EVS, Parameswaran TN, Singh R, Kalra A.
Effect of organic and inorganic fertilizers and Trichoderma harzianum
on patchouli (Pogostemon cablin) herb yield. J Med Aromat Plant Sci.
2010;32:50‐52.
126. Singh G, Hippalgaonkar KV. Influence of foliar applied kinetin on
growth and essential oil content of Patchouli (Pogostemon cablin
Benth.). Indian Perfum. 1993;37:167‐170.
127. de Jong AWK. Quelques observations sur les plantes à huiles
essentielles et sur leurs essences. Rec Trav Chim Pays‐Bas.
1912;30:211‐219.
128. Gogröf G. Untersuchungen über das ätherische Öl, die Sesquiterpene
und die Drüsenhaare von Pogostemon Patchouli Pell. Pharmazie.
1957;12:38‐51.
129. Yu J, Qi Y, Luo G, Duan HQ, Zhou J. Extraction and analysis of the
essential oil in Pogostemon cablin by enzymatic hydrolysis and
inhibitory activity against Hela cell proliferation. Zhong Yao Cai.
2012;35:796‐799.
130. Henderson W, Hart JW, How P, Judge J. Chemical and morphological
studies on sites of sesquiterpene accumulation in Pogostemon cablin
(Patchouli). Phytochemistry. 1970;9:1219‐1228.
131. Maeda E, Miyake H. Leaf anatomy of patchouli (Pogostemon patchouli)
with reference to the disposition of mesophyll glands. Jap J Crop Sci.
1997;66:307‐317.
132. Paschkis H. Contributions to a closer knowledge of some little
known leaves. I. Folia patchouli of commerce. Pharm J Trans.
1881;11:813‐815.
133. Solereder H. Bemerkenswerte anatomische Vorkommnisse bei
einigen Drogen. I. Die inneren haarartigen Sekretdrüsen des
Patschuliblattes. Arch Pharm. 1907;245:406‐414.
134. Sandes SS, Blank AF, Botanico MP, Blank MFA, Vasconcelos
JNC, Mendonca SAD. Estruturas secretoras foliares em
patchouli [Pogostemon cablin (Blanco) Benth.]. Scientia Plena.
2012;8‐059902:1‐6.
135. (a) Guo J, Yuan Y, Liu Z, Zhu J. Development and structure of internal
glands and external glandular trichomes in Pogostemon cablin. PLoS
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
45
One. 2013;8:e77862; (b) Rusydi A, Talip N, Latip J, Abdul Rahman R,
Sharif I. Morphology of trichomes in Pogostemon cablin Benth.
(Lamiaceae). Austr J Crop Sci. 2013;7:744‐749.
136. Dummond HH. Patchouli oil. Perfum Ess Oil Rec. 1960;51:484‐492.
137. Calkin RR, Jellinek JS. Perfumery ‐ Practice and Principles. New York:
John Wiley & Sons, Inc.; 1994;128‐129.
138. Holdsworth Haines W. The essential oil industry of Seychelles. Bull
Imp Inst. 1934;32:545‐558.
139. Anonymous. Patchouli Aceh ‐ A world‐class scent: Patchouli oil from
Aceh. Vol. 1, March, 2011;1‐16. Caritas Czech Republic, Indonesia,
https://sumarnidahlan.files.wordpress.com/2011/06/brosur‐caritas‐
st‐english.pdf
140. Harunsyah MY. Process design of patchouli oil distillation by varying
operating conditions to increase yields of patchouli oil, In Proceedings
of The 2nd Annual International Conference Syiah Kuala University 2012
& The 8th IMT‐GT Uninet Biosciences Conference, Banda Aceh,
2012;149‐153.
141. Anonymous. Standar Nasional Indonesia 06‐2385‐2006 ‐ Minyak
nilam, Badan Standardisasi Nasional, http://teknologihutan.fkt.ugm.
ac.id/userfiles/download/SNI_06‐2385‐2006‐Minyak_Nilam.pdf,
accessed on November 30, 2016.
142. Anonymous. Patchouli, http://vanaroma.com/patchouli/, accessed on
November 30, 2016.
143. Firmenich, Process for the stabilization of perfumes, US Patent
4,217,250, May 16, 1978.
144. Hua LJ, Chun LS. The utilization of molecular distillation for the
purification of some essential oils, In Proceedings of International
Congress of Essential Oils, Flavor & Fragrance and Cosmetic, Beijing,
1988;57‐62.
145. Sariadi. Pemurnian minyak nilam dengan proses adsorpsi
menggunakan bentonit. Jurnal Teknologi. 2012;12:100‐104.
146. Parry EJ. Refractive indices of essential oils. Chemist Druggist.
1910;77:52‐53.
147. Parry EJ. The refractive indices of essential oils. Chemist Druggist.
1910;76:178.
148. Treibs W. Der Patchouli‐alkohol, ein tricyclischer Azulen‐bildner.
Justus Liebigs Ann Chem. 1949;564:141‐151.
149. Šorm F, Dolejs L, Knessl O, Pliva J. On terpenes XVI. On a bicyclic
sesquiterpene and a new azulene from the oil of Pogostemon patchouli
P. Coll Czech Chem Commun. 1950;15:82‐95.
150. Büchi G, Erickson ER. Terpenes V. The structure of patchouly alcohol.
J Am Chem Soc. 1956;78:1262‐1263.
151. Dobler M, Dunitz JD, Gubler B, Weber HP, Büchi G, Padilla OJ. The
structure of patchouli alcohol. Proc Chem Soc London. 1963;383.
152. Bates PB, Slagel RC. Terpenoids. VI. β‐Bulnesene, α‐guaiene,
β‐patchoulene, and guaioxide in essential oils. Chem Ind.
1962;1715‐1716.
153. Wenninger JA, Yates RL, Dolinsky M. Sesquiterpene hydrocarbon
analysis as an aid in the characterization of commercial essential oils:
a study of patchouly, ylang ylang and gurjon balsam oils. Proc Sci Sect
Toilet Goods Assoc. 1966;44‐53.
154. Tsubaki N, Nishimura K, Hirose Y. Hydrocarbons in patchouli oil. Bull
Chem Soc Jpn. 1967;40:597‐600.
155. Mookherjee BD, Light KK, Hill ID. A study on the odor‐structure rela-
tionship of patchouli compound. In Essential Oils, Mookherjee BD,
Mussinan CJ (ed.). Allured Publishing Co.: Wheaton, 1981;247‐272.
156. Teisseire P, Maupetit P, Corbier B. Contribution à la connaissance de
l'huile essentielle de patchouli. Recherches. 1974;19:8‐35.
157. Proot M, Sandra P. High speed capillary GC on 10 m × 100 μm i.d.
FSOT columns. J High Res Chrom & Chrom Comm. 1986;9:618‐623.
158. Proot M. Capillaire kolommen met kleine inwendige diameter (≤ 100
μm) in gaschromatografie en superkritische vloeistofchromatografie.
Ph.D. Thesis, Universiteit Gent, Belgium, 1986.

46
159. Li F, Li C‐J, Ma J, et al. Four new sesquiterpenes from the stems of
Pogostemon cablin. Fitoterapia. 2013;86:183‐187.
160. Liu J‐l, Li X‐h, Peng C, et al. 4‐nor‐β‐Patchoulene sesquiterpenoids from
the essential oil of Pogostemon cablin. Phytochem Lett. 2015;12:27‐30.
161. Zhu H, Zhou Q‐M, Peng C, et al. Pocahemiketals A and B, two new
hemiketals with unprecedented sesquiterpenoid skeletons from
Pogostemon cablin. Fitoterapia. 2017;120:67‐71.
162. Zaizen K, Kusano Y, Kabayahi T, et al. Study about a novel odor‐active
compound in Patchouli oil, In Proceedings of 58th Symposium on the
Chemistry of Terpenes, Essential Oils and Aromatics (TEAC), Waka-
yama, 2014;184‐187.
163. Joulain D, Tabacchi R. Sesquiterpene hydrocarbons from patchouly
and patchoulol, In Proceedings of 31st International Symposium on
Essential Oils, Hamburg, Sept. 11, 2000, Oral communication L5.
164. Firmenich. Clearwood, http://www.firmenich.com/uploads/
files/ingredients/marketing‐sheet/perfumery/clearwood‐970953.pdf,
accessed on November 27, 2015.
165. Givaudan, Perfume compositions, World Patent 2017/050971,
September 24, 2015.
166. Anonymous. Essence de patchouly. Syndicat National des Fabricants
& Importateurs d'Huiles Essentielles & Produits Aromatiques
Naturels, 1955;83‐87.
167. Anonymous. Huile essentielle de patchouli [Pogostemon cablin (Blanco)
Benth.]. l'Association Française de Normalisation (AFNOR), Saint‐
Denis La Plaine, Saint‐Denis La Plaine. 2003;6.
168. Anonymous. Essential oils from East Africa. Bull Imp Inst.
1934;32:224‐226.
169. Anonymous. Essential oils from Seychelles. Bull Imp Inst.
1934;32:537‐539.
170. Chinese Pharmacopoeia Commission. Chinese Pharmacopoeia of the
People's Republic of China, Patchouli oil. Vol. I. 10th ed. Beijing: China
Medical Science and Technology Press; 2015;397.
171. Chinese Pharmacopoeia Commission. Chinese Pharmacopoeia of the
People's Republic of China, Pogostemonis Herba. Vol. I. 10th ed.
Beijing: China Medical Science and Technology Press; 2015;45.
172. Technical Committee ISO/TC54 Essential Oils. Aromatic natural raw
materials ‐ Vocabulary. ISO 9235, 1997.
173. Naves Y‐R, Mazuyer G. Natural Perfume Materials. New York: Reinhold
Publ. Corp.; 1947;82,136,282.
174. Naves Y‐R. Technologie et Chimie de Parfums Naturels ‐ Essences
Concrètes, Resinoïdes Huiles et Pommades aux Fleurs. Paris: Masson &
Cie.; 1974;266‐267.
175. Frister T, Hartwig S, Alemdar S, et al. Characterization of a recombi-
nant patchoulol synthase variant for biocatalytic production of
terpenes. Appl Biochem Biotechnol. 2015;176:2185‐2201.
176. Adams RP. Identification of Essential Oil Components by Gas Chroma-
tography / Mass Spectrometry. 4th edition. Carol Stream: Allured
Publishing Corporation; 2007.
177. Joulain D, König WA. The Atlas of Spectral Data of Sesquiterpene
Hydrocarbons. Hamburg: E.B.‐Verlag; 1998.
178. Hochmuth D. MassFinder 4 Software, http://massfinder.com/wiki/
MassFinder_4, accessed on April 29, 2017.
179. Milchard MJ, Clery R, Esdale R, et al. Application of gas‐liquid chroma-
tography to the analysis of essential oils: GLC fingerprint
chromatograms of five essential oils. Perfum Flavor. 2010;35:34,36‐
38,40,42.
180. Hasegawa Y, Tajima K, Toi N, Sugimura Y. An additional constituent
occurring in the oil from a Patchouli cultivar. Flavour Frag J.
1992;7:333‐335.
181. Murugan R, Mallavarapu GR, Padmashree KV, Rao RR, Livingstone C.
Volatile oil composition of Pogostemon heyneanus and comparison of
its composition with patchouli oil. Nat Prod Commun. 2010;5:1961‐
1964.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
182. Aisyah Y, Hastuti P, Sastrohamidjojo H, Hidayat C. Komposisi kimia
dan sifat antibakteri minyak nilam (Pogostemon cablin). Majalah
Farmasi Indon. 2008;19:151‐156.
183. Mookherjee BD, Trenkle RW, Wilson RA. New insights in the three
most important natural fragrance products: wood, amber, and musk, In
Proceedings of 12th International Congress of Flavours, Fragrances and
Essential Oils, Vienna, Fachzeitschriftenverlag GmbH, 1992;234‐247.
184. Mookherjee BD, Wilson RA. Recent progress in the chemistry of
character‐impact components of essential oils. In On Essential Oils,
Verghese J (ed.). Synthite: 1986; 282‐329.
185. Maurer B, Thomas AF. Alkaloids, bases and essential oils. Perfum
Flavor. 1994;19 (March/April):19‐27.
186. Hefendehl FW. The analytical evaluation of patchouli oil. Seifen, Oele,
Fette, Wachse. 1977;103:159‐161.
187. Welch SC, Chou C‐Y, Gruber JM, Assercq J‐M. Total synthesis of
(±)‐seychellene, (±)‐isocycloseychellene, and (±)‐isoseychellene. J Org
Chem. 1985;50:2668‐2676.
188. Gal H. Sur un homologue du camphre de Bornéo. CR Acad Sci.
1869;68:406‐408.
189. de Montgolfier J. Sur le camphre de patchouli. CR Acad Sci.
1877;84:88‐91.
190. von Soden H, Rojahn W. Ueber die Zusammensetzung des
Patchouliöles. Chem Ber. 1904;37:3353‐3355.
191. Bates PB, Slagel RC. Conversion of bulnesol to patchouli alcohol,
guaiol, and "δ‐guaiene". J Am Chem Soc. 1962;84:1307‐1308.
192. Büchi G, Erickson RE, Wakabayashi N. Terpenes. XVI. Constitution of
patchouli alcohol and absolute configuration of cedrene. J Am Chem
Soc. 1961;83:927‐938.
193. Teisseire P. The chemistry of patchouli oil. Rivista Ital Essenze Profumi.
1973;572‐591.
194. Peyron L. Sur quelques essences en provenance du Mato Grosso. Parf
Cosm Sav France. 1973;3:371‐378.
195. Wolff G, Ourisson G. Un nouveau sesquiterpene tricyclique: le
seychellene. Tetrahedron Lett. 1968;9:3849‐3852.
196. Wolff G, Ourisson G. Isolation and structure of seychellene.
Tetrahedron. 1969;25:4903‐4914.
197. Paknikar SK, Veeravalli J. Natural occurrence of δ‐patchoulene in
patchouli oil. Indian J Chem. 1980;19B:432.
198. Faraldos JA, Wu S, Chappell J, Coates RM. Doubly deuterium‐labeled
patchouli alcohol from cyclization of singly labeled [2‐2H1]farnesyl
diphosphate catalyzed by recombinant patchoulol synthase. J Am
Chem Soc. 2010;132:2998‐3008.
199. Welch SC, Gruber JM, Morrison PA. Total syntheses of (±)‐seychellene
and (±)‐cycloseychellene. J Org Chem. 1985;50:2676‐2681.
200. Huang A‐C, Sumby CJ, Tiekink ERT, Taylor DK. Synthesis of guaia‐
4(5)‐en‐11‐ol, guaia‐5(6)‐en‐11‐ol, aciphyllene, 1‐epi‐melicodenones
C and E, and other guaiane‐type sesquiterpenoids via the
diastereoselective epoxidation of guaiol. J Nat Prod. 2014;77:
2522‐2536.
201. Rakotonirainy O, Gaydou EM, Faure R, Bombarda I. Sesquiterpenes
from patchouli (Pogostemon cablin) essential oil. Assignment of the
proton and carbon‐13 NMR spectra. J Essent Oil Res. 1997;9:321‐327.
202. Rakotonirainy O, Gaydou EM, Faure R, Bombarda I. Errata of:
Sesquiterpenes from patchouli (Pogostemon cablin) essential oil.
Assignment of the proton and carbon‐13 NMR spectra. J Essent Oil
Res. 1997;9:608.
203. Mookherjee BD, Trenkle RW, Ledig WO. δ‐Patchoulene, an acid‐
catalyzed transformation product of patchouli alcohol. J Agric Food
Chem. 1974;22:771‐773.
204. Ishihara M, Tsuneya T, Uneyama K. Guaiane sesquiterpenes from
agarwood. Phytochemistry. 1991;30:3343‐3347.
205. Lawrence BM. The isolation of aromatic materials from natural plant
products. In A Manual on the Essential Oil Industry, Tuley de Silva K

VAN BEEK AND JOULAIN
(ed.). United Nations Inustrial Development Organization: Vienna,
1995; 57‐154.
206. Cornwell CP. Notes on the composition of Patchouli oil (Pogostemon
cablin (Blanco) Benth.). J Essent Oil Res. 2010;22:360‐364.
207. Saritas Y. Isolierung, Strukturaufklärung und stereochemische
Untersuchungen von sesquiterpenoiden Inhaltsstoffen aus
ätherischen Ölen von Bryophyta und höheren Pflanzen. Ph.D. Thesis,
University of Hamburg, 2000.
208. Buré CM, Sellier NM. Analysis of the essential oil of Indonesian patch-
ouli (Pogostemon cablin Benth.) using GC/MS (EI/CI). J Essent Oil Res.
2004;16:17‐19.
209. Blay G, Garcia B, Molina E, Pedro JR. Synthesis of (+)‐pechueloic acid
and (+)‐aciphyllene. Revision of the structure of (+)‐aciphyllene.
Tetrahedron. 2007;63:9621‐9626.
210. Srikrishna A, Pardeshi VH. Enantiospecific total synthesis of
aciphyllene. Tetrahedron. 2010;66:8160‐8168.
211. Terhune SJ, Hogg JW, Lawrence BM. Cycloseychellene, a new
tetracyclic sesquiterpene from Pogostemon cablin. Tetrahedron Lett.
1973;4705.
212. Dhekne VV, Paknikar SK. Identity of patchouli oil hydrocarbon E with
cycloseychellene. Indian J Chem. 1974;12:1016.
213. Willcott MR, Morrison PA, Assercq JM, Welch SC, Inners R.
Cycloseychellene: a structural revision. J Org Chem. 1981;
46:4819‐4820.
214. Wenninger JA, Yates RL, Dolinsky M. High resolution infrared spectra
of some naturally occurring sesquiterpene hydrocarbons. J AOAC.
1967;50:1313‐1335.
215. Wenninger JA, Yates RL. High resolution infrared spectra of some
naturally occurring sesquiterpene hydrocarbons. II. Second series. J
AOAC. 1970;53:949‐963.
216. Wallach O, Tuttle FE. II. Zur Charakteristik der Sesquiterpene. Justus
Liebigs Ann Chem. 1894;279:391‐397.
217. Gadamer J, Amenomiya T. Beitrage zur Kenntnis der Sesquiterpene
und Sesquiterpenalkohole. Arch Pharm. 1903;241:22‐42.
218. Semmler FW, Mayer EW. Notizen über einige Sesquiterpen Alkohole.
Chem Ber. 1912;45:1390‐1391.
219. Büchi G, MacLeod Jr. WD. Synthesis of patchouli alcohol. J Am Chem
Soc. 1962;84:3205‐3206.
220. Wienhaus H. Ester der Chromsäure. Ber Dtsch Chem Ges.
1914;47:322‐331.
221. Büchi G, MacLeod Jr. WD, Padilla O. J. Terpenes. XIX. Synthesis of
patchouli alcohol. J Am Chem Soc. 1964;86:4438‐4444.
222. Barton DHR, Belœil J‐C, Billion A, Boivin J, Lallemand J‐Y, Mergui S.
Functionalisation of saturated hydrocarbons. Oxidation of patchouli
alcohol by the 'Gif system': Isolation and organoleptic properties of
three new ketonic derivatives. Helv Chim Acta. 1987;70:273‐280.
223. Nicolaou KC, Snyder SA. Chasing molecules that were never
there: misassigned natural products and the role of chemical
synthesis in modern structure elucidation. Angew Chem Int Ed.
2005;44:1012‐1044.
224. Houghton PJ. The Chemistry of Valeriana. In Valerian ‐ The Genus
Valeriana, Houghton PJ (ed.). Harwood Academic Publishers:
Amsterdam, 1997; 21‐54.
225. Bicchi C, Sandra P, Schelfaut M, Verzele M. Studies on the essential oil
of Valeriana celtica L. J High Resol Chromatogr & Chromatogr Comm.
1983;6:213‐215.
226. Bos R, Woerdenbag HJ, Hendriks H, Friso Smit H, Wikström HV,
Scheffer JJC. Composition of the essential oil from roots and rhizomes
of Valeriana wallichii DC. Flavour Frag J. 1997;12:123‐131.
227. Kovacevic N, Pavlovic M, Menkovic N, Tzakou O, Couladis M.
Composition of the essential oil from roots and rhizomes of Valeriana
pancicii Halácsy & Bald. Flavour Frag J. 2002;17:355‐357.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
47
228. Verma RS, Verma RK, Padalia RC, Chauhan A, Singh A, Singh HP.
Chemical diversity in the essential oil of Indian Valerian (Valeriana
jatamansi Jones). Chem Biodiversity. 2011;8:1921‐1929.
229. IFF. Healingwood, http://lmrnaturals.iff.com/search.aspx, accessed
on November 29, 2016.
230. Roure Bertrand, Tricyclische alcohol, Dutch Patent 7211760, August
29, 1972.
231. Roure Bertrand, Alcool nor‐sesquiterpénique tricyclique sa prépara-
tion et ses applications, French Patent 2.151.522, September 1, 1971.
232. Teisseire P, Maupetit P, Corbier B, Rouillier P. norpatchoulénol, étude
chimique, structure et configuration absolue. Recherches (Paris).
1974;19:36‐61.
233. Oberhänsli WE, Schönholzer P. Structure cristalline et configuration
absolue d'un dérivé du Norpatchoulol 1a (IR, 3R 4aR, 6R, 8aS)
bromo‐3 hydroxy‐4a triméthyl‐5,5,8a méthano‐1,6 octahydro
naphtalénone‐2 (1H). Recherches (Paris). 1974;19:62‐68.
234. Bang L, Ourisson G, Teisseire P. Hydroxylation of patchoulol by
rabbits. Hemi‐synthesis of non‐patchoulenol, the odour carrier of
patchouli oil. Tetrahedron Lett. 1975;2211‐2214.
235. Teisseire P. Hydroxylations du patchoulol par voie biologique et
microbiologique. Bull Soc Chim France. 1980;II‐66 ‐ II‐70.
236. Pfau AS, Plattner P. Zur Kenntnis der flüchtigen Pflanzenstoffe. IV.
Über die Konstitution der Azulene. Helv Chim Acta. 1936;19:858‐879.
237. Hikino H, Ito K, Takemoto T. Sesquiterpenoids. XXVII. Structure of
pogostol. Chem Pharm Bull (Tokyo). 1968;16:1608‐1610.
238. Itokawa H, Morita H, Watanabe K, Iitaka Y. A new skeleton
sesquiterpenoid from Alpinia japonica (Thunb.) Miq. Chem Lett.
1984;451‐452.
239. Teisseire P. Industrial quality control of essential oils by capillary GC.
In Capillary Gas Chromatography in Essential Oil Analysis, Sandra P,
Bicchi C (ed.). Huethig: Heidelberg, 1987; 215‐258.
240. Fleischer TC, Waigh RD, Waterman PG. Pogostol O‐methyl ether and
artabotrol: Two novel sesquiterpenes from the stem bark of
Artabotrys stenopetalus. J Nat Prod. 1997;60:1054‐1056.
241. Weyerstahl P, Marschall H, Splittgerber U, Wolf D. Analysis of the
polar fraction of Haitian vetiver oil. Flavour Frag J. 2000;15:153‐173.
242. Stierle AA, Stierle DB, Goldstein E, et al. A novel 5‐HT receptor ligand
and related cytotoxic compounds from an acid mine waste extremo-
phile. J Nat Prod. 2003;66:1097‐1100.
243. Booker‐Milburn KI, Jenkins H, Charmant JPH, Mohr P. Synthesis of
the reported structure of pogostol and a total synthesis of (±)‐kessane
without the use of protecting groups. Org Lett. 2003;5:3309‐3312.
244. Amand S, Langenfeld A, Blond A, Dupont J, Nay B, Prado S. Guaiane
sesquiterpenes from Biscogniauxia nummularia featuring potent
antigerminative activity. J Nat Prod. 2012;75:798‐801.
245. Barra L, Schulz B, Dickschat JS. Pogostol biosynthesis by the endo-
phytic fungus Geniculosporium. ChemBioChem. 2014;15:2379‐2383.
246. ten Have R. De vaststelling van een relatieve configuratie van
pogostol met verschillende NMR‐technieken. M.Sc. Thesis,
Wageningen Agricultural University, 1992.
247. Givaudan, 1‐hydroxy‐octahydroazulenes as fragrances, European
Patent 2402301, September 29, 2010.
248. International Flavors and Fragrances (IFF), Hexahydro‐1,4,9,9‐
tetramethyl‐4,7‐methano‐azulenones, US Patent 3679750, June 10,
1968.
249. Trivedi B, Motl O, Herout V, Šorm F. On Terpenes. CLXIV. Composi-
tion of the oil from Cyperus rotundus L. Structure of patchoulenone.
Coll Czech Chem Commun. 1964;29:1675‐1688.
250. Thebtaranonth C, Thebtaranonth Y, Wanauppathamkul S, Yuthavong
Y. Antimalarial sesquiterpenes from tubers of Cyperus rotundus: struc-
ture of 10,12‐peroxycalamenene, a sesquiterpene endoperoxide.
Phytochemistry. 1995;40:125‐128.

48
251. Clery RA, Cason JRL, Zelenay V. Constituents of cypriol oil (Cyperus
scariosus R.Br.): N‐containing molecules and key aroma components.
J Agric Food Chem. 2016;64:4566‐4573.
252. Mookherjee BD, Trenkle RW, Wilson RA. New insights in the three
most important natural fragrance products: wood, amber and musk.
Indian Perfum. 1992;36:313‐332.
253. Givaudan, Patchoulione, US Patent 2942029, February 6, 1958.
254. Clery R. High‐impact odorants in essential oils. Flavour Frag J.
2010;25:117‐120.
255. Kaiser R. Meaningful Scents around the World. Olfactory, Chemical, Bio-
logical, and Cultural Considerations. Weinheim: Wiley‐VCH; 2006;63.
256. Huang A‐C, Burrett S, Sefton MA, Taylor DK. Production of the pep-
per aroma compound, (−)‐rotundone, by aerial oxidation of α‐guaiene.
J Agric Food Chem. 2014;62:10809‐10815.
257. Huang A‐C, Sefton MA, Taylor DK. Comparison of the formation of
peppery and woody sesquiterpenes derived from α‐guaiene and α‐
bulnesene under aerial oxidative conditions. J Agric Food Chem.
2015;63:1932‐1938.
258. Su J‐H, Chiang MY, Wen Z‐H, Dai C‐F, Hsu C‐H, Sheu J‐H.
Sesquiterpenoids from the Formosan soft coral Sinularia leptoclados.
Chem Pharm Bull. 2010;58:250‐253.
259. Kiuchi F, Matsuo K, Ito M, Qui TK, Honda G. New sesquiterpene
hydroperoxides with trypanocidal activity from Pogostemon cablin.
Chem Pharm Bull. 2004;52:1495‐1496.
260. Toyota M, Koyama H, Mizutani M, Asakawa Y. (−)‐ent‐Spathulenol
isolated from liverworts is an artefact. Phytochemistry.
1996;41:1347‐1350.
261. Klein E, Rojahn W. Isolierung, Struktur und Synthese von Dhelwangin.
Tetrahedron Lett. 1969;2279‐2280.
262. Luo JP, Liu YP, Feng YF, Guo XL, Cao H. Two chemotypes of
Pogostemon cablin and influence of region of cultivation and harvesting
time on volatile oil composition. Acta Pharmacol Sin. 2003;38:307‐310.
263. Luo J, Guo X, Feng Y. Constituents analysis on volatile oil of
Pogostemon cablin from different collection time cultivated in Hainan.
Zhong Yao Cai. 2002;25:21‐23.
264. He Y, Deng C, Xiong L, Qin S, Peng C. Transcriptome sequencing
provides insights into the metabolic pathways of patchouli alcohol
and pogostone in Pogostemon cablin (Blanco) Benth. Genes Genomics.
2016;1031‐1039.
265. Ouyang P, Liu Y, Wang Y, Mo X, Zeng S. Aging and/or tissue‐specific
regulation of patchoulol and pogostone in two Pogostemon cablin
(Blanco) Benth. cultivars. Physiol Plant. 2016;158:272‐283.
266. Büchi G, Goldman IM, Mayo DW. Structures of two alkaloids from
patchouli oil. J Am Chem Soc. 1966;88:3109‐3113.
267. van der Gen A, van der Linde LM, Witteveen JG. Synthesis of
guaipyridine and some related sesquiterpene alkaloids. Rec Trav Chim
Pays‐Bas. 1972;91:1433‐1440.
268. Maurer B, Hauser A. New pyridine derivatives from essential oils.
Chimia. 1992;46:93‐95.
269. Anonymous. Semi‐annual Report of Schimmel & Co. Miltitz.
1904;68‐71.
270. Chaintreau A, Cicchetti E, David N, et al. Collaborative validation of
the quantification method for suspected allergens and test of an auto-
mated data treatment. J Chromatogr A. 2011;1218:7869‐7877.
271. de Rijke D, Traas PC, ter Heide R, Boelens H, Takken HJ. Acidic com-
ponents in essential oils of costus root, patchouli and olibanum.
Phytochemistry. 1978;17:1664‐1666.
272. Masada Y. Analysis of Essential Oils by Gas Chromatography and Mass
Spectrometry. New York: John Wiley & Sons; 1976;74‐76.
273. Morris ET. Patchouli ‐ Faszination eines Duftes. Dragoco Report.
1983;05:128‐133.
274. Hollo J, Kurucz E, Torky AA, Biacs P. Application de la CCM à l'étude
des huiles volatiles. Plant Food Hum Nutr (Qualitas Plantarum et
Materiae Vegetabiles). 1968;16:51‐62.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
275. Hikino H, Ito K, Aota K, Takemoto T. Synthesis of cyperene,
cyperotundone, and patchoulenone. Chem Pharm Bull. 1968;16:43‐51.
276. International Flavors and Fragrances (IFF), Tetracylic alcohols, US
Patent 3991123, September 17, 1975.
277. Zhou L, Xu M, Yang C‐R, Wang Y‐F, Zhang Y‐J. New patchoulol‐type
sesquiterpenoids from Pogostemon cablin. Helv Chim Acta.
2011;94:218‐223.
278. Nakahara S, Kumatani K, Kameoka H. Acidic compounds in patchouli
oil. Phytochemistry. 1975;14:2712‐2713.
279. International Flavors and Fragrances (IFF), Fragrance materials
containing cis‐2‐N‐pentylcyclopropane‐1‐carboxylic acid, US Patent
3926860, December 17, 1974.
280. Albuquerque ELD, Lima JKA, Souza FHO, et al. Insecticidal and
repellence activity of the essential oil of Pogostemon cablin against
urban ants species. Acta Trop. 2013;127:181‐186.
281. Wu L, Wu Y, Guo Q, Li S, Zhou K, Zhang J. Comparison of genetic
diversity in Pogostemon cablin from China revealed by RAPD, morpho-
logical and chemical analyses. J Med Plants Res. 2011;5:4549‐4559.
282. Sundaresan V, Singh SP, Mishra AN, et al. Composition and comparison
of essential oils of Pogostemon cablin (Blanco) Benth. (Patchouli) and
Pogostemon travancoricus Bedd. var. travancoricus. J Essent Oil Res.
2009;21:220‐222.
283. Tsai Y‐C, Hsu H‐C, Yang W‐C, Tsai W‐J, Chen C‐C, Watanabe T.
α‐Bulnesene, a PAF inhibitor isolated from the essential oil of
Pogostemon cablin. Fitoterapia. 2007;78:7‐11.
284. Bunrathep S, Lockwood GB, Songsak T, Ruangrungsi N. Chemical con-
stituents from leaves and cell cultures of Pogostemon cablin and use of
precursor feeding to improve patchouli alcohol level. ScienceAsia.
2006;32:293‐296.
285. Silva MAS, Ehlert PAD, Ming LC, Marques MOM. Composition and
chemical variation during daytime of constituents of the essential oil
of Pogostemon patchouli pellet leaves. Acta Hortic. 2004;629:145‐147.
286. Nguyen XD, Leclercq PA. Chemical composition of patchouli oil from
Vietnam. J Essent Oil Res. 1989;1:99‐100.
287. Akhila A, Nigam MC. GC‐MS analysis of the essential oil of
Pogostemon cablin (patchouli oil). Fitoterapia. 1984;55:363‐365.
288. Bacci L, Lima JKA, Araujo APA, et al. Toxicity, behavior impairment,
and repellence of essential oils from pepper‐rosmarin and patchouli
to termites. Entomol Exp Appl. 2015;156:66‐76.
289. Singh M, Sharma S, Ramesh S. Herbage, oil yield and oil quality of
patchouli [Pogostemon cablin (Blanco) Benth.] influenced by irrigation,
organic mulch and nitrogen application in semi‐arid tropical climate.
Ind Crops Prod. 2002;16:101‐107.
290. Singh R, Divya S, Awasthi A, Kalra A. Technology for efficient and
successful delivery of vermicompost colonized bioinoculants in
Pogostemon cablin (patchouli) Benth. World J Microbiol Biotechnol.
2012;28:323‐333.
291. Zeng Z, Tan L, Meng S, Zhang H. Study on the chemical constituents
and fingerprint of Pogostemon cablin from three culture varieties. Chin
J Anal Chem. 2006;34:1249‐1254.
292. Feng Y, Guo F, Luo J. GC‐MS analysis of volatile oil of Herba
Pogostemonis collected from Leizhou county. Zhong Yao Cai.
1999;22:241‐243.
293. Luo J, Feng Y, Guo X, Li X. GC‐MS analysis of volatile oil of Herba
Pogostemonis collected from Gaoyao county Zhong Yao Cai.
1999;22:25‐28.
294. Sugimura Y, Padayhag BF, Ceniza MS, et al. Essential oil production
increased by using virus‐free patchouli plants derived from meri-
stem‐tip culture. Plant Pathol. 1995;44:510‐515.
295. Kubeczka K‐H, Formácek V. Essential Oil Analysis by Capillary Gas
Chromatography and Carbon‐13 NMR Spectroscopy. New York: Wiley;
2002;233‐238.
296. Wossa SW, Rali T, Leach DN. Volatile chemical constituents of
Patchouli (Pogostemon cablin (Blanco) Benth.: Labiateae) from three

VAN BEEK AND JOULAIN
localities in Papua New Guinea. Papua New Guinea J Agric Forest Fish.
2006;49:49‐54.
297. Srinivas SR. Atlas of Essential Oils. New York: Published by the author;
1986;628‐630.
298. Williams DG. The Chemistry of Essential Oils. Second Edition. Port
Washington: Micelle Press; 2008;198‐199.
299. Silva‐Filho SE, Wiirzler LAM, Cavalcante HAO, et al. Effect of patch-
ouli (Pogostemon cablin) essential oil on in vitro and in vivo
leukocytes behavior in acute inflammatory response. Biomed
Pharmacother. 2016;84:1697‐1704.
300. Hardoyono F, Iswanto BH, Triyana K. Comparative analysis of feature
extraction methods in the clustering of electronic nose response
correlated with GC/MS analysis, In Proceedings of AIP Conf Proc,
American Institute of Physics, 2016, 1755‐170006: 1‐6.
301. de Rapper S, Kamatou G, Viljoen A, van Vuuren S. The in vitro antimi-
crobial activity of Lavandula angustifolia essential oil in combination
with other aroma‐therapeutic oils. J Evid Based Complementary Altern
Med. 2013;852049:1‐10.
302. Orchard A, Sandasi M, Kamatou G, Viljoen A, van Vuuren S. The
in vitro antimicrobial activity and chemometric modelling of 59 com-
mercial essential oils against pathogens of dermatological relevance.
Chem Biodiversity. 2017;14‐e1600218:1‐18.
303. Prawoto AA. Early yield and economical study of Pogostemon cablin as
intercrop in young cocoa (Theobroma cacao L.). Indon Agric Res Abstr.
2010;27:6.
304. Hu H, Peng J, Huang S, Wu L, Zhu B, Xuan Y, Yang D. Study on puri-
fication technology of patchouly oil with molecular distillation.
Zhongguo Zhongyao Zazhi. 2004;29:320‐322, 379.
305. Sugimura Y, Toi N. Yield and quality of essential oils from aroma crops.
Jpn J Crop Sci. 1991;60:324‐331.
306. Adhavan P, Kaur G, Princy A, Murugan R. Essential oil nanoemulsions
of wild patchouli attenuate multi‐drug resistant gram‐positive, gram‐
negative and Candida albicans. Ind Crops Prod. 2017;100:106‐116.
307. Hu LF, Li SP, Cao H, et al. GC‐MS fingerprint of Pogostemon cablin in
China. J Pharm Biomed Anal. 2006;42:200‐206.
308. Liu YP, Luo JP, Feng YF, Guo XL, Cao H. DNA profiling of Pogostemon
cablin chemotypes differing in essential oil composition. Acta Pharm
Sinica. 2002;37:304‐308.
309. de Groot AC, Schmidt E. Essential Oils ‐ Contact Allergy and Chemical
Composition. Boca Raton: CRC Press; 2016;643‐652.
310. Milchard MJ, Clery R, DaCosta N, Esdale R, et al. Application of gas‐
liquid chromatography to the analysis of essential oils: fingerprints
of 12 essential oils. Perfum Flavor. 2004;29:28‐36.
311. Lamparsky D. "Fingerprints" in essential oil analysis: Aim, results, and
correlations with sensory values. In Capillary Gas Chromatography in
Essential Oil Analysis, Sandra P, Bicchi C (ed.). Huethig: Heidelberg,
1987; 155‐213.
312. Tissot E, Rochat S, Debonneville C, Chaintreau A. Rapid GC‐FID quan-
tification technique without authentic samples using predicted
response factors. Flavour Frag J. 2012;27:290‐296.
313. de Saint Laumer J‐Y, Cicchetti E, Merle P, Egger J, Chaintreau A.
Quantification in gas chromatography: Prediction of flame ionization
detector response factors from combustion enthalpies and molecular
structures. Anal Chem. 2010;82:6457‐6462.
314. Cachet T, Brevard H, Chaintreau A, et al. IOFI recommended practice
for the use of predicted relative‐response factors for the rapid quan-
tification of volatile flavouring compounds by GC‐FID. Flavour Frag J.
2016;31:191‐194.
315. Cicchetti E, Merle P, Chaintreau A. Quantitation in gas chromatogra-
phy: usual practices and performances of a response factor
database. Flavour Frag J. 2008;23:450‐459.
316. van den Dool H. Standardisation of gas chromatographic analysis of
essential oils. Ph.D. Thesis, University of Groningen, 1974.
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
49
317. van Beek TA. Natural Products Analysis: Quo Vadis?, In Proceedings of
Advances in Natural Products Analysis, Grasse, France, Pole PASS, Nov.
23, 2010.
318. Filippi J‐J, Belhassen E, Baldovini N, Brevard H, Meierhenrich UJ.
Qualitative and quantitative analysis of vetiver essential oils by
comprehensive two‐dimensional gas chromatography and compre-
hensive two‐dimensional gas chromatography/mass spectrometry. J
Chromatogr A. 2013;1288:127‐148.
319. Anonymous. Guidelines for the quantitative gas chromatography of
volatile flavouring substances, from the Working Group on Methods
of Analysis of the International Organization of the Flavor Industry
(IOFI). Flavour Frag J. 2011;26:297‐299.
320. Bicchi C, Liberto E, Matteodo M, et al. Quantitative analysis of essen-
tial oils: a complex task. Flavour Frag J. 2008;23:382‐391.
321. van Beek TA, van Veldhuizen A, Lelyveld GP, Piron I, Lankhorst PP.
Quantitation of bilobalide and ginkgolides A, B, C and J by means
of nuclear magnetic resonance spectroscopy. Phytochem Anal.
1993;4:261‐268.
322. Analytical Methods Committee. Application of gas ‐ liquid chromatogra-
phy to the analysis of essential oils. Part XII. Determination of patchouli
alcohol in oil of patchouli. Analyst (London). 1987;112:1315‐1318.
323. Yu Z‐X, Wang L‐J, Zhao B, et al. Progressive regulation of sesquiter-
pene biosynthesis in Arabidopsis and patchouli (Pogostemon cablin)
by the miR156‐targeted SPL transcription factors. Mol Plant.
2015;8:98‐110.
324. Kang SS, Kim JS, Chi HJ, Won DH. Isolation and quantitative determi-
nation of patchouli alcohol from Pogostemon cablin Benth. Korean J
Pharmacog. 1998;29:18‐21.
325. Zhao Z, Lu J, Leung K, Chan CL, Jiang Z‐H. Determination of
patchoulic alcohol in Herba Pogostemonis by GC‐MS‐MS. Chem
Pharm Bull. 2005;53:856‐860.
326. Hybertson BM. Solubility of the sesquiterpene alcohol patchoulol in
supercritical carbon dioxide. J Chem Eng Data. 2007;52:235‐238.
327. Li X‐C, Zhang Q‐W, Yin Z‐Q, Zhang X‐Q, Ye W‐C. Preparative separa-
tion of patchouli alcohol from patchouli oil using high performance
centrifugal partition chromatography. J Essent Oil Res. 2011;23:19‐24.
328. Wang X‐g, Zou Y‐f, Wang Y‐s. GC‐MS assay for the determination of
pogostone in essential oil of herba pogostemonis. Chin J Pharm Anal.
2005;25:582‐584.
329. Nikiforov A, Jirovetz L, Buchbauer G, Raverdino V. GC‐FTIR and GC‐
MS in odor analysis of essential oils. Mikrochim Acta. 1988;2:193‐198.
330. Ruan C, Xu G, Lu X, Hua R, Kong H, Xiao K, Yang Q. Quality evaluation
of volatile oils of traditional Chinese medicines by comprehensive
2‐dimensional gas chromatography (GC × GC). Chromatographia.
2003;57:S/265‐S/270.
331. Wu J, Lu X, Tang W, Kong H, Zhou S, Xu G. Application of compre-
hensive two‐dimensional gas chromatography‐time‐of‐flight mass
spectrometry in the analysis of volatile oil of traditional Chinese med-
icines. J Chromatogr A. 2004;1034:199‐205.
332. Edwards M, McGregor L, Barden D. Detailed characterization of
essential oils by flow‐modulated GC×GC with tandem ionization
TOF‐MS. The Column. 2016;12‐16:15‐21.
333. Sonwa MM. Isolation and structure elucidation of essential oil constit-
uents ‐ comparative study of the oils of Cyperus alopecuroides, Cyperus
papyrus, and Cyperus rotundus. Ph.D. Thesis, University of Hamburg,
2000.
334. Betts TJ. Evaluation of a "Chirasil‐Val" capillary for the gas chromatog-
raphy of volatile oil constituents, including sesquiterpenes in patchouli
oil. J Chromatogr A. 1994;664:295‐300.
335. Schmidt E, Wanner J. Adulteration of essential oils. In Handbook of
Essential Oils ‐ Science, Technology, and Applications, 2nd ed.
Başer KHC, Buchbauer G (ed.). CRC Press: Boca Raton, 2016; 707‐745.
336. Hirschsohn E. Testing ethereal oils. J Chem Soc, Abstr. 1896;70:B223.
337. Anonymous. Scientific and other notes on essential oils. Semi‐annual
Report of Schimmel & Co. 1917;April‐October:33.

50
338. Wei A, Shibamoto T. Antioxidant activities and volatile constituents of
various essential oils. J Agric Food Chem. 2007;55:1737‐1742.
339. Teisseire P. Quelques aspects de la chromatographie en phase
gazeuse dans l'industrie des matières odorantes utilisées en
parfumerie. Recherches. 1969;17:37‐43.
340. Kaloustian J. Contrôles chromatographiques. In La Connaissance des
Huiles Essentielles Qualitologie et Aromathérapie, Kaloustian J,
Hadji‐Minaglou F (ed.). Springer: Paris, 2012;70‐72.
341. Dann AE, Sait S, Swan DFK, Stewart I. Rapid quality control analysis of
essential oils using a capillary gas chromatography technique, In
Proceedings of IXth International Congress of Essential Oils, Singapore,
The Essential Oil Association of Singapore, 1983;194.
342. Andrade BFMT, Barbosa LN, da Silva Probst I, Fernandes Jr. A.
Antimicrobial activity of essential oils. J Essent Oil Res. 2014;26:34‐40.
343. Han X, Beaumont C, Stevens N, Parker TL. Chemical composition
analysis and in vitro biological activities of ten essential oils in human
skin cells. Biochemie Open. 2017;5:1‐7.
344. Tang XT, Wang YY, Tang YL, Chen D‐F, Method of utilizing the PTS
gene and anti‐sense ADS to increase patchouli alcohol content in
Artemisia annua L., US Patent 2011/0300547, June 3, 2011.
345. Kraft P. Woody pretzels: spirocycles from Vetiver to Patchouli and
Georgywood. Chem Biodiversity. 2008;5:970‐999.
346. Anonis DP. Woody notes in perfumery: patchouly oil, absolute and
aroma chemicals: part I. Perfum Flavor. 2006;31:36‐39.
347. Arctander S. Perfume and Flavor Materials of Natural Origin. Elizabeth:
Arctander; 1960.
348. Lis‐Balchin M. Aromatherapy with essential oils. In Handbook of Essen-
tial Oils ‐ Science, Technology, and Applications, 2nd ed. Başer KHC,
Buchbauer G (ed.). CRC Press: Boca Raton, 2016;619‐653.
349. Hasegawa T. Separation of odor constituents by microscale fractional
bulb‐to‐bulb distillation. In Distillation: Advances from Modeling to
Applications, Zereshki S (ed.). InTech: Rijeka, 2012; 199‐210.
350. Fráter G, Lamparsky D. Synthetic products. In Perfumes Art Science and
Technology, Müller PM, Lamparsky D (ed.). Elsevier: London, 1991;566.
351. Dragoco Gerberding & Co., C‐8‐substituted 1,5‐dimethyl‐
bicyclo[3.2.1]octane‐8‐ols, US Patent 4,582,945, July 21, 1982.
352. Kastner D. Patchulol oder Norpatschulenol? Riechst Arom Kosmet.
1977;77:79‐80.
353. Teisseire P. Recent developments in the chemistry and biochemistry
of patchoulol, In Proceedings of 7th International Congress of Essential
Oils, Kyoto, October 1977, Japan Flavor Fragrance Manufactures'
Assoc., 1979;315‐319.
354. Petrzilka M, Ehret C. Natural Products. In Perfumes Art Science and
Technology, Müller P M, Lamparsky D (ed.). Elsevier: London,
1991;521.
355. Näf F, Decorzant R, Giersch W, Ohloff G. A stereocontrolled access to (±)‐,
(−)‐, and (+)‐patchouli alcohol. Helv Chim Acta. 1981;64:1387‐1397.
356. Nikiforov A, Jirovetz L, Buchbauer G. (±)‐Patchoulol, der dominante
Geruchsträger des Patchouliöls (singapur.) Monatsh Chem.
1986;117:1095‐1098.
357. Firmenich, Utilisation de certains dérivés sesquiterpéniques en tant
qu'agents parfumants, Swiss Patent 289452, July 23, 1974.
358. Roure Bertrand Dupont, Bicyclic alcohols, a process for their prepara-
tion and odoriferous compositions containing them, European Patent
0074543, September 15, 1981.
359. Firmenich, Cyclic tertiary alcohols and their use as perfuming ingredi-
ents, US Patent 5268356, November 25, 1991.
360. Weyerstahl P, Gansau C, Marschall H. Structure‐odor correlation. Part
XVIII. Partial structures of patchoulol with bicyclo[2.2.2]octane skele-
ton. Flavour Frag J. 1993;8:297‐306.
361. Leffingwell D. Chirality and odor perception ‐ Scents of precious
woods. Chim Oggi / Chem Today. 2006;24, supplement:36‐38.
362. Wood C, Siebert TE, Parker M, Capone DL, Elsey GM, Pollnitz AP,
Eggers M, Meier M, Vössing T, Widder S, Krammer G, Sefton MA,
10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN
Herderich MJ. Spice up your life ‐ The rotundone story, In Proceedings
of the 12th Weurman Symposium, Interlaken, Switzerland, Zürcher
Hochschule für Angewandte Wissenschaften, 2008;483‐485.
363. Wood C, Siebert TE, Parker M, Capone DL, Elsey GM, Pollnitz AP,
Eggers M, Meier M, Vössing T, Widder S, Krammer G, Sefton MA,
Herderich MJ. From wine to pepper: rotundone, an obscure sesquiter-
pene, is a potent spicy aroma compound. J Agric Food Chem.
2008;56:3738‐3744.
364. Leffingwell J. (−)‐Rotundone, an important odor active constituent of
agarwood (Oud), is now FEMA GRAS, http://leffingwell.com/
rotundone.htm, accessed on March 17, 2017.
365. Pompizzi R. Furanfettsäuren als Vorläufer von Aromastoffen. Ph.D.
Thesis, ETH, Zürich, 1999.
366. KAO‐Corporation, Fragrant patchouli oil and a perfume composition
comprising said patchouli oil, European Patent 0409089, July 20,
1989.
367. Deguerry F, Pastore L, Wu S, Clark A, Chappell J, Schalk M. The
diverse sesquiterpene profile of patchouli, Pogostemon cablin, is corre-
lated with a limited number of sesquiterpene synthases. Arch Biochem
Biophys. 2006;454:123‐136.
368. Firmenich, Sesquiterpene synthases from patchouli, World Patent
2005/052163, November 19, 2004.
369. Firmenich, Method for producing patchoulol and 7‐epi‐slinene, World
Patent 2011/141855, May 10, 2010.
370. Daviet L, Schalk M. Biotechnology in plant essential oil production:
progress and perspective in metabolic engineering of the terpene
pathway. Flavour Frag J. 2010;25:123‐127.
371. Bomgardner MM. The sweet smell of microbes. Chem Engin News.
2012;90:25‐29.
372. Rastogi SC, Heydorn S, Johansen JD, Basketter DA. Fragrance
chemicals in domestic and occupational products. Contact Dermatitis.
2001;45:221‐225.
373. de Groot AC, Schmidt E. Essential Oils, Part 1: Introduction. Dermati-
tis. 2016;27:39‐42.
374. Belsito D, Bickers D, Bruze M, et al. A toxicologic and dermatologic
assessment of cyclic and non‐cyclic terpene alcohols when used as
fragrance ingredients Food Chem Toxicol. 2008;46:S1‐S71.
375. Bhatia SP, Letizia CS, Api AM. Fragrance material review on patchouli
alcohol. Food Chem Toxicol. 2008;46:S255‐S256.
376. Oser BL, Carson S, Oser M. Toxicological tests on flavouring matters.
Food Cosmet Toxicol. 1965;3:563‐569.
377. Mitchell JC. Contact hypersensitivity to some perfume materials.
Contact Dermatitis. 1975;1:196‐199.
378. Nakayama H, Matsuo S, Hayakawa K, Takahashi K, Shigematsu T,
Ota S. Pigmented cosmetic dermatitis. Int J Dermatol. 1984;23:
299‐305.
379. Frosch PJ, Johansen JD, Menne T, et al. Further important sensitizers
in patients sensitive to fragrances. II. Reactivity to essential oils. Con-
tact Dermatitis. 2002;47:279‐287.
380. Uter W, Schmidt E, Geier J, Lessmann H, Schnuch A, Frosch P. Contact
allergy to essential oils: current patch test results (2000‐2008) from
the Information Network of Departments of Dermatology (IVDK).
Contact Dermatitis. 2010;63:277‐283.
381. Steinberg DC. The EU fragrance allergens. Cosmet Toiletries.
2012;127:434,436,438,440.
382. Geier J, Uter W. Diagnostic workup of fragrance allergy. Hautarzt.
2015;66:674‐679.
383. European Commission Scientific Committee on Consumer Safety.
Opinion on fragrance allergens in cosmetic products. 2011;91‐92.
384. European Union. Directive 2003/15/EC. Off J Eur Union. 2003;
L 66:26‐35.
385. International Fragrance Association (IFRA). IFRA Standard ‐
Isophorone. 43rd Amendment, October 14, Brussels. 2009;1‐2.
 10991026, 2018, 1, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ffj.3418 by Nat Prov Indonesia, Wiley Online Library on [22/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
VAN BEEK AND JOULAIN 51

386. European Committee for Standardization. Methods for analysis of 390. Hasegawa T, Yoshitome K, Fujihara T, Santoso M, Aziz MA. Character-
allergens ‐ Quantification of suspected fragrance allergens in istic changes in the aroma profile of patchouli depending on
consumer products ‐ Step 1: GC analysis of ready‐to‐inject sample. manufacturing process. J Oleo Sci. 2017;66:863‐869.
BS EN 16274, 2012.
387. European Chemicals Agency. EC number: 939‐227‐3, https://echa. SUPPOR TI NG INF ORMATI ON
europa.eu/fr/registration‐dossier/‐/registered‐dossier/14973?p_auth=
Additional Supporting Information may be found online in the
ZjH9g9vA, accessed on January 1, 2017.
supporting information tab for this article.
388. Yoshitome K, Hasegawa T, Fujihara T, Santoso M, Aziz MA. Change in
odor of patchouli according to dyring process, In Proceedings of 17th
International Symposium on Olfaction and Taste, Yokohama, June 5‐9,
2016, Poster P‐061.
How to cite this article: van Beek TA, Joulain D. The essential
389. Yoshitome K, Hasegawa T, Fujihara T, Santoso M, Aziz MA,
oil of patchouli, Pogostemon cablin: A review. Flavour Fragr J.
Yamada H. Study on aroma profile of patchouli depending on
manufacturing process, In Proceedings of 60th TEAC, Abashiri, 2018;33:6–51. https://doi.org/10.1002/ffj.3418
2016;311‐313.