VOLATILE CONSTITUENTS OF GENIPAP 583

Copyright © 2005 John Wiley & Sons, Ltd. Flavour Fragr. J. 2005; 20: 583–586

FLAVOUR AND FRAGRANCE JOURNALFlavour Fragr. J. 2005; 20: 583–586Published online 25 April 2005 in Wiley InterScience (www.interscience.wiley.com). DOI: 10.1002/ffj.1491

Volatile constituents of genipap (Genipa americana L.)fruit from Cuba

Jorge Pino,1 Rolando Marbot2 and Carlos Vazquez3

1 Instituto de Investigaciones para la Industria Alimenticia, Carretera del Guatao km 31/2, La Habana 19200, Cuba2 Centro Nacional de Genética Médica, Instituto Superior de Ciencias Médicas, La Habana, Cuba3 Jardín Botánico Nacional, La Habana, Cuba

Received 18 August 2003; Revised 1 March 2004; Accepted 20 April 2004

ABSTRACT: Volatile constituents were isolated from genipap (Genipa americana L.) fruit by simultaneous steam

distillation–solvent extraction according to Likens–Nickerson. Compounds were identified by capillary GC-MS. Ninety-

eight compounds were identified in the aroma concentrate, of which hexadecanoic acid, octadecanoic acid, tetradecanoic

acid, linalool and limonene were found to be the major constituents. Carboxylic acids, with their acidic and harsh notes,

should play an important role in the flavor of the fruit. Copyright © 2005 John Wiley & Sons, Ltd.

KEY WORDS: Genipa americana; genipap; fruit volatiles; GC-MS; hexadecanoic acid; octadecanoic acid; tetradecanoic acid;

linalool; limonene

* Correspondence to: J. Pino, Instituto Investigaciones para la Industria

Alimenticia, Carretera del Guatao km 31/2, La Habana, Cuba.

E-mail: agu@iiia.edu.cn

Introduction

The genipap, locally called ‘genipa’, is the species

Genipa americana L., probably native to Antilles or

South America. This fruit belongs to the botanical

family Rubiaceae. It is an evergeeen tree growing to a

height of about 10–20 m. The fruits are globose, 5–8 cm

in diameter, with a yellowish-gray outside. The pulp of

the ripe fruit is juicy, acidic and harsh. It is usually eaten

as juices, jams and liquors.1 These aromatic properties led

us to investigate its composition in order to determine the

respective influence of the volatile constituents.

To date the aroma of G. americana fruit has been the

subject of only one previous study from fruits grown

in Brazil.2 In this study, 35 compounds were detected

by GC-MS after 4 h Likens–Nickerson distillation with

dichloromethane as solvent and, using GC-AEDA, the

short chain acids were found to be responsible for the

pungent and acid notes, while the ethyl esters presented

the most intense and characteristic notes of this fruit. In

this study we have, therefore, examined the volatile con-

stituents of the genipap fruit grown in Cuba.

Experimental

Materials

Samples of ripe fruits were collected from the National

Botanic Garden in Havana, in May 2001, from a single

specimen, at the same time. A voucher specimen of

the plant is deposited in the herbarium of the National

Botanic Garden in Havana.

Isolation of the volatile constituents

After addition of an internal standard (methyl unde-

canoate, 2 mg), pulp (200 g) was blended with distilled

water (800 ml) and simultaneously distilled and extracted

for 90 min in a Likens–Nickerson microapparatus

with 25 ml of diethyl ether (previously redistilled and

checked as to purity). The volatile concentrate was

dried over anhydrous sodium sulfate and concentrated

to 0.6 ml in a Kuderna–Danish evaporator with a 12 cm

Vigreux column, and then to 0.2 ml with a gentle nitro-

gen stream.

Analysis of the volatile constituents

The extract was analyzed by GC using a Hewlett-Packard

6890 gas chromatograph equipped with a flame ionization

detector (FID). The separations were performed using a

SPB-5 column (30 m × 0.25 mm i.d., 0.25 µm) with an

oven temperature program of 60 °C (2 min), increasing at

4 °C/min to 250 °C (20 min). The carrier gas was helium

with a flow-rate of 1 ml/min. The temperature of the

injector and detector was 250 °C. The injection was made

in the split mode (1:10 ratio). Linear retention indices

were calculated against those of n-paraffins.3 These condi-

tions were used for quantitative analysis by the internal

standard method. The recovery of the method was deter-

mined by the standard addition technique applied to a

584 J. PINO ET AL.

Copyright © 2005 John Wiley & Sons, Ltd. Flavour Fragr. J. 2005; 20: 583–586

sample. The analytes α-pinene, limonene, 1-hexanol, (Z)-

3-hexenol, benzaldehyde, methyl benzoate and linalool

were added at two different concentrations. The average

recoveries were about 88–102% and their relative stand-

ard deviations were lower than 10%.

GC-MS analyses were performed with a Hewlett-

Packard series 6890 gas chromatograph equipped with a

HP-5973 mass-selective detector. The chromatographic

conditions were the same as those described for GC-FID.

The detector was operated in electron impact mode

(70 eV) at 230 °C. Detection was performed in the scan

mode between 30 and 400 amu.

Compounds were identified by comparing their spectra

with those of the Wiley library or our IDENT library and

also by comparison of their GC Kovats indices with those

of standard compounds and data from the literature.4,5

Results and discussion

The volatile constituents of fresh genipap fruit were

obtained by simultaneous steam distillation–solvent

extraction and analyzed by GC and GC-MS using fused

silica capillary columns. Table 1 summarizes the qualita-

tive and quantitative analyses of the fruit volatiles accord-

ing to order of elution on the SPB-5 column. The yield

of total volatiles, estimated by the addition of a measured

amount of internal standard to the pulp, was appro-

ximately 18 mg/kg of fruit pulp.

Ninety-eight constituents were identified as volatile

components of genipap, 81 of them reported for the

first time in the fruit. A rough survey of the chemical

classes represented in this fruit flavour was as follows:

carboxylic acids comprise the largest class of volatiles

(69.6%); the composition of the other classes of com-

pounds was: terpenoids, 10.1%; alcohols, 8.7%; esters,

6.6%; aldehydes and ketones, 1.0%; and others, 3.9%.

The major constituents found in genipap flavor were

hexadecanoic acid (9.35 mg/kg), octadecanoic acid

(1.34 mg/kg), tetradecanoic acid (1.16 mg/kg), linalool

(0.91 mg/kg) and limonene (0.62 mg/kg).

In the previous investigation of genipap volatiles,2 low

weight molecular carboxylic acids and their methyl and

ethyl esters predominated in relative area. Although

carboxylic acids were also detected as main compounds,

many terpenoids were identified for the first time. Other

Table 1. Volatile constituents of genipap fruit

Retention Criteria of ConcentrationCompound indexa identificationb (mg/kg)

Acetaldehydec 435 A <0.01

Ethanolc 500 A 0.20

1-Propanolc 568 A 0.03

Diacetylc 612 A <0.01

Ethyl acetatec 615 A 0.22

Isobutanolc 622 A 0.21

Benzenec 646 A 0.02

3-Methylbutanalc 650 A <0.01

1-Butanolc 668 A 0.06

2,3-Pentanedionec 700 A <0.01

Methyl isobutyrate 716 A <0.01

3-Methyl-1-butanolc 736 A 0.48

Pyridinec 752 A 0.01

Ethyl isobutyratec 756 A <0.01

(Z)-2-Penten-1-olc 767 C <0.01

Toluenec 773 A 0.01

3-Methyl-2-buten-1-ol 779 A 0.07

Ethyl lactatec 789 A <0.01

Ethyl butyratec 800 A <0.01

Hexanalc 801 A 0.09

2-Furfural 807 A 0.02

Methyl 2-methylbutyrate 810 A <0.01

3-Methyl-1-pentanolc 812 C <0.01

2-Methylbutyric acid 826 C 0.03

(Z)-3-Hexen-1-olc 857 A 0.07

1-Hexanolc 867 A 0.07

Styrenec 880 C <0.01

p-Xylenec 884 A 0.04

2-Heptanonec 889 A <0.01

Heptanalc 900 A <0.01

(E,E)-2,4-Hexadienalc 909 A 0.01

Methyl 2-methylbutyratec 918 A <0.01

α-Pinenec 939 A 0.01

Propylbrenzenec 957 A <0.01

(E)-2-Heptenalc 960 B <0.01

VOLATILE CONSTITUENTS OF GENIPAP 585

Copyright © 2005 John Wiley & Sons, Ltd. Flavour Fragr. J. 2005; 20: 583–586

Table 1. (Continued )

Retention Criteria of ConcentrationCompound indexa identificationb (mg/kg)

Benzaldehyde 961 A <0.01

1,2,4-Trimethylbenzenec 966 A <0.01

1-Heptanolc 969 A <0.01

β-Pinene 980 A 0.05

1-Octen-3-olc 981 A 0.15

Myrcenec 991 A 0.02

3-Octanolc 993 A 0.02

α-Terpinenec 1018 A <0.01

p-Cymenec 1025 A 0.02

Limonene 1031 A 0.62

Benzyl alcohol 1032 A 0.15

Cyclohexyl acetatec 1043 C 0.18

2-Phenylacetaldehydec 1046 A <0.01

(E)-β-Ocimenec 1050 A <0.01

(E)-2-Octenalc 1054 B 0.01

γ -Terpinenec 1062 A 0.09

Acetophenonec 1065 A 0.01

Benzofuranc 1072 C 0.22

Terpinolenec 1088 A 0.01

Methyl benzoate 1091 A 0.01

Linalool 1098 A 0.91

Nonanalc 1102 A 0.02

2-Phenylethanolc 1110 A 0.07

Methyl octanoate 1126 A <0.01

p-Toluyl alcoholc 1135 C <0.01

trans-o-Coumaric acidc 1139 C 0.17

Methyl phenylacetatec 1145 A 0.03

2-Methylbenzofuranc 1168 C <0.01

Dimethyl glutaconatec 1187 C 0.60

Methyl salicylatec 1190 A 0.02

Decanalc 1204 A <0.01

Benzothiazolec 1221 A <0.01

2,3-Dihydrobenzofuranc 1224 C <0.01

Methyl 2-methylbenzoatec 1226 A <0.01

Methyl 3-methylbenzoatec 1235 A <0.01

Isobutyrophenonec 1239 C <0.01

Phenylpropanoic acidc 1258 C <0.01

2-Vinyl-2,3-dihydrobenzofuranc 1279 C 0.02

Safrolec 1285 A 0.02

Indolec 1288 A <0.01

(E,Z)-2,4-Decadienalc 1291 B 0.01

4-Vinylguaiacolc 1323 C 0.34

Methyl 2,5-dihydrobenzoatec 1324 A 0.05

Methyl decanoatec 1327 A <0.01

Eugenolc 1356 A <0.01

γ-Nonalactonec 1359 A 0.01

Geranic acidc 1359 C <0.01

Methyl p-hydroxyhydrocinnamatec 1360 C <0.01

(Z)-β-Damascenonec 1361 A 0.02

Decanoic acidc 1380 A 0.08

δ-Cadinenec 1524 A <0.01

Dodecanoic acidc 1581 A 0.08

Methyl tetradecanoate 1725 A 0.02

Benzyl benzoatec 1762 A <0.01

Tetradecanoic acid 1782 A 1.16

Pentadecanoic acidc 1881 A 0.14

Trimethyl 1,3,5-benzenetricarboxylatec 1892 C 0.03

Methyl hexadecanoate 1926 A 0.03

Hexadecanoic acid 1981 A 9.35

Methyl octadecanoatec 2128 A <0.01

Linoleic acidc 2132 A 0.53

Octadecanoic acidc 2180 A 1.34

a Retention index on SPB-5 column.b The reliability of the identification proposal is indicated by the following: A, mass spectrum and Kovats index agreed

with standards; B, mass spectrum and Kovats index agreed with literature data; C, mass spectrum agreed with mass

spectral database.c Reported for the first time.

586 J. PINO ET AL.

Copyright © 2005 John Wiley & Sons, Ltd. Flavour Fragr. J. 2005; 20: 583–586

differences were due to high molecular weight carboxylic

acids predominating in our investigation.

Some compounds present, e.g. furfural and some

furanes, could be degradation products of ascorbic acid

and sugars.6,7 Nevertheless, the concentrate was found, on

appropriate redilution with water, to possess the charac-

teristic genipap aroma.

As sensory evaluations were not carried out in this

study it is difficult to determine which of the components

may contribute more to the flavor of genipap. It is our

opinion that this fruit does not have easily identifiable

flavor impact compounds, but among them the carboxylic

acids, with their acidic and harsh notes, play an important

role.

References

1. Martin FW, Cambell CW, Ruberté RM. Tropical Edible Fruits of

the Tropics: an Inventory. USDA, Agriculture Research Service:Washington, DC, 1987.

2. Borges ES, Rezende CM. J. Essent. Oil Res., 2000; 12: 71–74.

3. Majlat P, Erdos Z, Takacs J. J. Chromatogr., 1974; 91: 89–110.

4. MacLafferty FW, Stauffer DB. The Wiley/NBS Registry of Mass

Spectral Data. Wiley: New York, 1989.5. Adams RP. Identification of Essential Oil Components by Gas

Chromatography/Mass Spectroscopy. Allured: Carol Stream, IL,1995.

6. Shaw PE, Tatum JH, Berry RE. J. Agric. Food Chem., 1968; 16:979–982.

7. Tatum JH, Shaw PE, Berry RE. J. Agric. Food Chem., 1969; 17:38–40.