FLAVOUR AND FRAGRANCE JOURNAL, VOL.

1, 149-153 (1986)

Additional Neutral Volatiles from Litchi (Litchichinensis Sonn.)

Fruit
0.Friihlich and P. Schreier
Lehrstuhl f i r Lebensmittelchemie, Universitat Wiirzburg, A m Hubland, 0-8700Wurzburg, FRG

The volatiles from fresh litchi (Litchi chinemis Sonn.) fruit pulp were separated by standard controlled
high-vacuum distillation with subsequent solvent extraction (pentane-dichloromethane, 2: 1) and silica gel liquid
chromatography pre-separation using a pentane-diethyl ether gradient. Polar fruit constituents were separated by
chloroform extraction and subsequent liquid-liquid distribution. The concentrates were analysed by capillary gas
chromatography (HRGC) and coupled HRGC techniques, i.e. on-line capillary gas chromatography-mass
spectrometry using electron impact and chemical ionization mode (HRGC-EI/CI-MS) as well as capillary gas
chromatography-Fourier transform infrared spectroscopy (HRGC-FTIR). These techniques revealed the
occurrence of 34 aroma components, not previously reported as litchi fruit constituents.

KEY WORDS Litchi chinensis Sonn. y o m a components Capillary gas chromatography Capillary gas
chromatography-mass spectrometry Capillary gas chromatography-Fourier transform infrared
spectroscopy

INTRODUCTION supplied by Hameico GmbH, Wiirzburg, were

Litchi chinensis Sonn., a native of South China
and actually cultivated there as well as in various
other countries, such as Australia, Brazil, Burma,
India, Japan, New Zealand, South Africa and the
U.S.A., is especially known for the attractive
flavour of its fruit, which has been described as
‘rose- and fruity-floral cherry-like’.’
In 1980 Johnston et al.’ detected 42 compon-
ents in a dichloromethane extract of litchi fruit
pulp and attributed the floral odour to 2-
phenylethanol and its derivatives. Four years
later, Toulemonde and Beauverd3 identified 89
volatiles from the headspace of litchi and con-
sidered different methyl ethers of various alcohols
as well as limonene, rose oxide, nonanal, decanal,
citronellyl and geranyl alcohols and acetates to be
responsible for the fruity-floral and citrus notes of
litchi fruit. In this paper, we report several addi-
tional aroma constituents, which were not
described in these previous publications.
EXPERIMENTAL
Fruits
Fresh, ripe litchi fruits (L. chinensis Sonn.),
imported from the Republic of South Africa and
analysed within two days of arrival.
Sample Preparation
A total of 4.8 kg of diluted fruit pulp was
obtained from 3.1 kg of total fruit after removal
of skin and the kernels, addition of 2.5 1 of dis-
tilled water and homogenization by a Braun blen-
der. After adding internal standards (butyl-
benzene, 0.53 mg/kg; ethyl (E)-hept-4-enoate,
0.55 mg/kg; 2-methylhexan-3-01, 0.48 mg/kg),
the diluted pulp (pH 4.7) was subjected to high-
vacuum distillation with subsequent solvent
extraction as well as direct CHC13 extraction/
liquid-liquid distribution.
High-vacuum Distillation-Solvent Extraction
High-vacuum distillation was carried out at
45”C/0.1 mbar over 3 h using two dry-
ice/methanol (-28°C) and two liquid nitrogen
cooled traps.4 The four traps were thawed and the
combined aqueous distillates (1.5 1) were extrac-
ted with pentane-dichloromethane (2: 1) over
24 h.’ The extracts were dried over anhydrous
sodium sulphate and carefully concentrated to
approximately 0.2 ml by using a Vigreux
column (45°C).

0882-5 734/86/040 149-05$05.00 Received 2 December 1986

0 1986 by John Wiley & Sons, Ltd.
150 0.FROHLICH A N D P. SCHREIER

Column Chromatography 30 ml/min H2 and 300 ml/min air, respectively.

The detector temperature was kept at 250°C.
The concentrated extracts obtained from distil-Volumes of 0.3 pl were injected.
lates were fractionated on silica gel 60 (Merck), Results of qualitative analyses were verified by
activity grade 11, by using a pentane-diethyl ether
comparison of HRGC retention, mass spectral,
gradient.6 Cooled (11-13°C) glass columns, and, in part, FTIR vapour-phase data with those
2.0 cm i.d. x 40 cm, were used. The elution rate of authentic reference substances. Quantitative
was 60 ml/h and three fractions were separated. HRGC determinations were carried out by stan-
Fraction I was eluted with 300 ml of pentane, dard controlled calculations using a Hewlett-
fraction 11 was obtained by eluting with 300 ml of Packard 3388 A laboratory data system without
diethyl ether-pentane (1:9, v/v), and fraction 111consideration of calibration factors, i.e. F = 1.00
was collected by eluting with 300 ml of diethyl for all compounds.
ether. All eluates were dried over anhydrous
sodium sulphate and concentrated to 0.2 ml (for
HRGC and HRGC-EI/CI-MS study) and to
0.05 ml (for HRGC-FTIR analysis), respec- Capillary Gas Chromatography-Mass
tively. Spectrometry (HRGC-MS)
A Varian Aerograph 1440 gas chromatograph
equipped with a Carlo Erba water-cooled on-
Separation of Polar Components column injection system was combined by direct
After addition of 1 1 of 0.2 M phosphate buffer coupling to a Finnigan MAT 44 mass spectro-
(pH 7.8) 600 g of litchi fruit pulp (skin and ker- meter. A J & W DB Wax fused silica capillary
nels removed) was homogenized by a Braun column (30 m, 0.32 mm i.d., film thickness,
blender and centrifuged at 18 000 x g . The 0.25 pm) connected to a 3 m uncoated piece of
supernatant was extracted three times each with fused silica capillary column as the 'retention
300 ml of CHC13, the extracts were dried over gap" was used. The conditions were as follows:
anhydrous sodium sulphate and concentrated in a temperature, isothermal for 2.5 min at 40°C and
vacuum rotary evaporator (30°C). Remaining then from 40 to 220" at 5"C/min; carrier gas flow
solvent was removed by a nitrogen flow and the rate, 2.5 ml/min He; temperature of ion source
oily residue was extracted twice with 10 ml of and all connection parts, 200°C; electron energy,
distilled water each time. The combined aqueous 70 eV; cathodic current, 0.8 mA. CI-MS was
phase was extracted with 30ml of pentane, the performed under the following conditions: reag-
solvent phase was discarded, and the aqueous ent gas, methylpropane; electron energy, 140 eV;
phase was extracted three times with 50ml of cathodic current, 0.5 mA; CI-box pressure,
CHC13 each time. The combined CHC13 extracts 450 Itbar.
were dried over anhydrous sodium sulphate and
concentrated by vacuum rotary evaporation
(30°C) to 0.1 ml (= fraction IV). Capillary Gas Chromatography -FTIR
Spectroscopy (HRGC-FTIR)

Capillary Gas Chromatography (HRGC)
A Carlo Erba Fractovap 4160 gas chromato-
graph with FID equipped with a J & W fused
silica DB Wax capillary column (30 m, 0.25 mm
i.d., film thickness = 0.25 pm) with a 3 m
uncoated fused silica capillary precolumn as the
'retention gap" was used. On-column injection
with an air-cooled injection system was em-
ployed. The temperature programme was
isothermal for 3 min at 50°C and then from 50 to
250°C at 4"C/min. The flow rates for the carrier
gas were 2.5 ml/min He, for the make-up gas
30 ml/min N2, and for the detector gases
HRGC-FTIR analysis was carried out with a
Nicolet 20 SXB system interfaced by a Dani 6500
gas chromatograph equipped with FID. A J & W
fused silica DB Wax capillary column (30m,
0.32 mm i.d., film thickness 0.25 pm) was used.
Total sample injection mode using programmed
temperature vaporization (PTV) (4O-24O0C,
0.1 min) was performed. The temperature pro-
gramme was 3 min isothermal at 50°C and then
from 50 to 250°C at 4"C/min. Light pipe and
transfer line were held at 250°C; He (2.5 ml/min)
was employed as carrier gas. Vapour phase FTIR
spectra were recorded from 400 to 4000cm-'
with a resolution of 8 cm-'.
 Table 1. Additional volatiles of litchi fruit and their chromatographic and spectroscopic data
~ ~

Compound Identified in’ R,b m/zc Vapour phase FTIR (cm-’)d

Alcohols vO-H vC-0 vC- H

2-(2-butoxyethoxy)ethanol 1v 1754 45-57-41-75-56-59-72-87 -
3-methylthiopropan-1-01 IV 1668 61-58-57-106-47-49-48-59 -
2-methylbut-3-en-2-01 111 1026 71-43-59-41-42-60-53-58 3639 1173 2982
3-methylbut-2-en-1-01 I11 1313 71-41-43-53-86-68-67-42 3657 1003 2930
(E)-hex-3en- 1-01 I11 1353 41-67-82-55-69-42-70-54 -
(Z)-hex-3en-l-ol 111 1371 41-67-55-42-82-69-57-54 3668 1049 2970
(E)-hex-2-en-1-01 111 1391 57-41-44-43-82-67-56-55 3661 1085 2969
heptan-1-01 I11 1458 56-70-43-41-55-42-69-57 -
benzyl alcohol 111 1864 79-77-108-107-51-50-78-106
3.7-dimethylocta-1,5-diene-3,7-diol 1v 1927 82-43-71-67-41-55-83-53 3643 1053 2979
3.7-dimethylocta-2.5-diene-1,%dial IV 2195 43-55-41-137-109-67-83-81 -
p-cymen-8-01 111 1820 43-135-109-124-91-81-65-150 -
p-menth-8-ene-1,2-diol IV 2272 71-43-67-41-69-58-55-82 -
8-bisabolol I11 2128 41-82-93-43-69-55-72-83 -
Carbonyl compounds vC=O vC-H
3-methylbutan-2-one 111 920 43-86-42-44-41 -45 1732 2978
cyclopentanone III 1144 55-41-84-56-42-44-54-53 1766 2975
2-methylhexan-3-one 11 1068 43-71-41-42-114-55-70-44 -
octan-3-one I1 1206 43-57-42-41-71-72-99-56 -
butyrophenone I1 1745 105-77-51-148-104-120-106-76 -
(E)-hex-2-enal I1 1212 41-55-69-42-57-83-43-56
octanol I1 1268 43-44-41-57-56-55-42-84
citronella1 I1 1456 41-69-55-43-95-56-67-53
Erhers vC-O-C vC-H
linalyl ethyl ether I1 1229 71-93-99-41-43-69-80-55 -
geranyl ethyl ether I1 1552 41-69-68-57-67-43-93-71 1112 2931
benzyl ethyl ether I1 1421 91-92-79-107-77-65-51-135 1118 2982/3038
1,2-dimethoxybe.nzene I1 1689 138-77-95-123-41-52-65-51 -
Esters vC=O v--C--O-C vC-H
neryl acetate 11 1717 69-41-43-68-93-42-67-80 1759 1231 2930
ethyl hexanoate I1 1215 88-43-60-70-99-61-41-73 1754 1172 2941
3-methylbutyl benzoate I1 1880 70-105-77-55-51-42-41-43 -
Hydrocarbons
8-phellandrene I 1200 93-77-91-136-79-9441-9s
perillene I 1387 41-69-81-82-53-150-51-135
a-acoradiene I 1641 119-93-105-121-91-55-67-147
a-himachalene I 1625 93-94-105-119-79-41-69-91

Miscellaneous vc=o vc-0 v0- H

2,5-dimethyl-l-4-hydroxy-3(2H)-furanone IV 1995 43-57-85-128-56-45-44-55 1781 1114 3578
“Fraction number; refer to experimental section for details.
bLinear retention index* determined on*DB Wax.
T h e eight most intense peaks are represented.
dThe most typical bands are represented.
152 0. FROHLICH AND P. SCHREIER

RESULTS AND DISCUSSION is-as an already-known constituent of litchi

By means of HRGC, HRGC-MS and
HRGC-FTIR 14 alcohols, 8 carbonyl compunds,
4 ethers, 3 esters, 4 hydrocarbons and one
furanone derivative were identified for the first
time in litchi (L. chinensis Sonn.) fruit. The
chromatographic and spectroscopic data of these
components are represented in Table 1. The
quantitative distribution of main constituents of
litchi fruit is outlined in Table 2. As shown in
Table 2, the methyl-branched C4-alcohols quan-
titatively occupy a special place among the major
litchi volatiles. Some substances have been
already described previously.293
Among the alcohols a series of C6-compounds
was detected, which are well-known as 'secon-
dary' volatiles formed after cell disruption from
unsaturated fatty acids due to lipoxygenase-hyd-
roperoxide lyase a ~ t i v i t yIt
. ~is obvious that these
volatiles could not be found during the headspace
analysis of the whole fruit carried out previously
by Toulemonde and B e a ~ v e r d . ~
In fraction IV, 3-methylthiopropan- 1-01 was
identified together with several monoterpene
diols. This thioether alcohol is a well-known
flavour compund derived from methionine
metabolism.' The acyclic monterpene diols listed
in Table 1 can be regarded as possible precursors
of nerol oxide, which was also identified, but
flavour-not represented here. In fraction IV,
some further components were found showing
MS data of monterpene diols, but structural
elucidation could not be achieved because of lack
of authentic reference material.
/?-Bisabolol, (1). originally isolated from the
essential oil of cotton buds, has been described as
exhibiting an apple blossom-like odour." From its
structure, biogenetic relations to the sesquiterpe-
nes a-acoradiene, (2), and a-himachalene, (3),
1 2 3
both newly identified in litchi fruit in the hyd-
rocarbon fraction I, can be considered. A series of
acoradienes has been recently detected in the
essential oil of Acorus calamus.'2 Perillene, first
isolated from Perilfu frufescen~,'~ could be regar-
ded as an oxidation product of (E)-ocimene, iden-
tified previously3and also in our present study in
litchi fruit.
The occurrence of rnonoterpene ethers has
already been pointed out in the previous investi-
g a t i o n ~ .In
~ addition to the described methyl

Table 2. Quantitative distribution of main constituents of litchi fruit volatiles

30-200 &kg 200-1000 &kg 1000 pglkg

of pullp of pullp of p u l p

hexan-I -01 2-methylpropan-1-01 3-methylbutan-1-01

(E)- hex-2-en-.l-ol. 3-methylbutyl-3-en-1-01
oft-1-en-3-01 3-methylbutyl-2-en-1-01'
8-bisabolol' citronellol
geraniol

3-hydroxybu tan-2-one

limonene
p-cymene
terpinolene
a-acoradiene'

3-methylbutyl acetate

geranylethyl ether.

'Standard controlled HRGC determination without consideration of calibration factors,

i.e. F = 1.00 for all compounds.
'Described for the first time as litchi fruit volatiles (cf. Table 1).
 VOLATILES FROM LITCHI FRUIT
ethers, two monoterpene ethyl ethers were newly
found together with benzyl ethyl ether. Informa-
tion about the natural Occurrence of monoter-
penoid ethers is scarce. Because of the careful
sample preparation without any use of ethanol,
chemical (artefact) formation has to be excluded.
Among the esters and carbonyls several volati-
les were identified, which have been commonly
found in various fruit flavours. Finally, 2,5-
dirnethyl-Chydroxy-3(2H)-furanone has already
been identified in several fruits. It was isolated at
the same time as a constituent of pineappleI4 and
of ~trawberry'~ and, with its powerful caramel-
like odour, occupies a central position among the
industrially used flavour substances.
Among the newly identified volatiles there are
compounds exhibiting fruity-floral and spicy
odours, which can be assumed to contribute to the
litchi fruit flavour. However, analysis was not
carried out to evaluate their individual sensory
contribution.
Acknowledgernenr-C. Kahre's excellent operation of
HRGC-FTIR analyses is gratefully acknowledged.
153
REFERENCES
1. K. Herrmann, Exorische Lebensmitrel, Springer, Berlin,
Heidelberg, New York (1983).
2. J. C . Johnston, R. C . Welch and G. L. K. Hunter,J. Agric.
Food Chem., 28, 859 (1980).
3. B. Toulemonde and D. Beauverd, in Progress in Flavour
Research, ed. J. Adda, p. 533, Elsevier, Amsterdam
(1984).
4. H. Idstein and P. Schreier, J. Agric. Food. Chem., 33,
138 (1985).
5. F. Drawert and A. Rapp, Chromatographia, 1, 466
(1968).
6. H. Idstein, W. Herres and P. Schreier, 1. Agric. Food
Chem., 32, 383 (1984).
7. K. Grob and R. Miiller, 1. Chromarogr., 244, 185 (1982).
8. H. van den Dool und P.D. Kratz,J. Chromarogr., 11,463
(1963).
9. T. Galliard, in Biochemistry of Wounded Plant Tissues,
ed. G. Kahl, p. 155, W. de Gruyter, Berlin, New York,
1978.
10. P. Schreier, F. Drawert, A. Junker, H. Barton and G.
Leupold, Z . Lebensm. Unrers.-Forsch., 162,279 (1976).
11. J. P. Minyard, A. C . Thompson and P.A. Hedin, 1. Org.
Chem., 39, 909 (1968).
12. G. Mazza, 1. Chromatogr., 328, 179 (1985).
13. H.Kondo and H. Suzuki, Chem. Eer., 69, 2459 (1936).
14. J. 0. Rodin, C . M . Himel, R. M. Silverstein, R. W. Leeper
and W. A. Gortner, 1. Food. Sci., 30, 230 (1965).
15. B. Willhalm, M. Stoll and A. F. Thomas, Chem. Ind.
(London), 1629 (1965).