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Procedia FoodScience Science 00 (2011) ProcediaFood 1 (2011) 404000000 407

11th International Congress of Engineering and Food (ICEF11)

Identification of volatile components isolated from indigenous fruits of Mozambique: Maphilwa (Vangueria infausta)
Rui Tique Raicea*.b, Ingegerd Sjoholma, Jos da Cruz Franciscob, Bjorn Bergenstahla
a

Department of Food Technology, Engineering and Nutrition; Lund University, Sweden b Department of Chemistry, Eduardo Mondlane University; Maputo, Mozambique

Abstract The aim of this work is to find the optimal combination of solvents which can extract the aroma components and identify the aroma profile of maphilwa (Vangueria infausta). Different extraction solvents like pentane, diethyl ether, ethanol and water were studied. Geraniol was used as internal standard added to the ethanol. Gas Chromatography was used to investigate the aroma components profile in maphilwa (V. infausta). The aroma compounds appears at relatively high values of retention time 15.1, 20.2 and 35.3 minutes for peak a, b and c respectively, reflecting the different boiling points.
Published by Elsevier Selection and/or peer-review and/or under responsibility of 11th International Congress on of Engineering 2011 2011 Published by B.V. Elsevier Ltd. Selection peer-review under responsibility ICEF11 and Food (ICEF 11) Executive Committee.

Executive Committee Members

Keywords: GC; aroma components; volatile; indigenous fruits of Mozambique; maphilwa (Vangueria infausta).

1. Introduction Substantial quantities of perishable fruits and vegetables with high nutritional value [1] grow in Mozambique as well as everywhere in souther part of Africa. Most of these fruits and vegetables (commonly known by local names) are available during the seasons of harvest, usually from January to May. Some examples of fruits are rubber vine or mavungwa (Landolpia kirki), maanica (Ziziphus mauritiana), mapsinsha (Salacia kraussi), false medlar or maphilwa (Vangueria infausta), kaffir orange or Kwazulu Natal orange or massala (Strychnos spinosa) [2]. These fruits can be considered as a possible additional source of food especially during periods when there are restrictions of other products. Most fruits have high content of micronutrients and fibres. But overall the fruits have to have a good taste when fresh and after processing. Volatile compounds are also present in these fruits. However, there are very few reports about the identification and quantification of volatile components from the indigenous fruits. The volatile chemicals present in natural leaves, flowers and fruits have been widely used in
* Corresponding author: Rui T. Raice. Department of Food Technology, Engineering and Nutrition, Lund University - Sweden; Tel.: +46 46 222 8310; fax: +46 46 222 9517. E-mail address: raice.rui@food.lth.se).

2211601X 2011 Published by Elsevier B.V. Selection and/or peer-review under responsibility of 11th International Congress on Engineering and Food (ICEF 11) Executive Committee. doi:10.1016/j.profoo.2011.09.062

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aromatherapy since ancient times [3]. In this study we investigate the fruit Vangueria infausta. The fresh V.infausta common called maphilwa in Ronga, (local language), it is sweet and has a taste like wild apple. It is brown-orange when ripe and have an orange flesh. The size is about 3-5 cm in diameter and it contains 2-3 seeds. The aim of this study is to find the optimal combination of solvents which can extract the aroma components of V. infausta in a reproducible manner. We have studied different extraction solvents such as pentane, diethyl ether, ethanol and investigated the effect of water. Geraniol was used as internal standard added to the ethanol. 2. Material and Methods Ripe fruits of maphilwa, (Vangueria infausta) were collected in Marracuene, 30 km north from Maputo (Mozambique). Matured fruits free from defects were selected, washed, sealed in vacuum bag and frozen at minus 18C. At the moment of analyses the fruits were thawed, boiled for 5 minutes, manually peeled and the pulp separated from seeds. The aromas are made more mobile by adding etanol in the ratio 1:1 to the sample. The internal standard, geraniol, was added to the etanol (0.04% V/V). The sample preextraction continued during 24 h and followed by filtration. The aromas were extracted using a hydrophobic solvent. Different extraction solvents have been evaluated: diethyl ether (D), pentane (P) and mixtures thereof. The ratio of the preextract to solvent was kept 1:1. GC analysis of extract was performed on a Agilent Technologies 6890N gas chromatography equipped with DB-225 column (30mx0,25mm) film 0,25 m with stationary phase containing 50% cyanopropylphenyldietylpolysiloxane. The oven temperature was programmed to increase from 50C to 200C during 5 min and than from 200C to 240C during 40 minutes. The injector and detector temperatures were 300C. The carrier gas was helium, pressure 55,5 kPa, and a split ratio of 40:1 flowing at 24,1ml/min. The hydrogen flow was 30 ml/min [4, 5]. The tentative concentration of the aroma compounds was calculated using the following equation: (1) Where C is a concentration of aroma component (given in mg/g), mIS is a mass of internal standard, mS is a mass of the sample, Api is a area of the peak of the sample, and ApIS is a area of the peak of internal standard. 3. Sample preparation 3.1. Isolation of volatile components by solvent extraction Two fruits (15 g) of maphilwa (Vangueria infausta) sample were manually peeled and crashed in small pieces (about 10 m) and added 15 ml of Ethanol containing 0,04% V/V of geraniol as internal standard. The mixture was continuously steered for 12 hours on any magnetic rotator. Then was filtered and added 25 ml of mixture of pentane and diethyl ether (1:1) and kept for extraction during 2 hours under steering. The organic phase (supernatant) was removed and introduced in a 1,5 ml GC vial, and crimped for GC analysis.

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4. Results and Discussion Volatile components of maphilwa (Vangueria infausta) were extracted using pentane and diethyl ether. The aroma extracts were found distributed, on up phase to possess the characteristic maphilwa aroma. The use of small amount of sample and shortening the boiling time during the thawing has great influence to broking the enzymatic reaction. In fact, very few reports are available for identification of aroma in proposed fruit but regarding similar research for essential oil composition in different materials since leaves as well as fruits has been used GC and HLPC and extraction methods [6, 7]. In this present work was used ethanol which, due to its hydrophilic character, cause more penetration into pulp fruit tissue and lead to more decomposition of cell content thus allow a good extractability of aroma components. Other hand the ethanol due to its miscibility, remove water from stigma matrix system and denature enzyme (-glucodidase) which hinders degradation during extraction time that could bring influences to the qualitative and quantitative results. As aroma has a non polar structure the use of non polar solvent alone or in combination such the mixture of pentane and diethyl ether (1:1) would improve the extraction of aromatic compounds. The extraction was held under continuous steering by magnetic apparatus that provide more liberation of volatile compounds into ethanol which increase the extraction yield of volatile components. In order to see the extraction reproducibility of the method, three replicates of different samples were similarly extracted. The solvent peaks appear at very low retention time while the aroma compounds appears at relatively high values of retention time 15,1, 20,2, and 35,3 min for peak a, b and c respectively, due to increasing of boiling point of the aroma compounds. The peak of the internal standard, (geraniol b.p=2290C) appear at 25 min.

Fig. 1. Peaks of aroma compounds isolated from three different replicates of maphilwa (Vangueria infausta) samples

5. Conclusions The aroma compounds appears at relatively high values of retention time 15,1, 20,2 and 35,3 min for peak a, b and c respectively, reflecting the different boiling points. The trial of extraction with combination of Diethyl ether (D), pentane (P) in ratio 1:1 give repeatable results on GC. Combining use of two extraction solvents of different boiling point and polarity enhance the extraction efficiency GC.. GC is a flexible method that can be used to characterize and semi quantify the volatile components isolated from maphilwa (Vangueria infausta.)

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The identification of the aroma constituent in maphilwa (V. infausta) are intended to be performed using GC-MS which remain to be the next step of this work. Acknowledgments This work was done with financial support from the SIDA-SAREC program that is fitfully thanks. The authors wish to thanks also the villagers of Marracuene region by guiding on collecting of the fruits used for this study. References
[1] Amorteifio, J O; Mosase, M O. The chemical composition of selected indigenous fruits of Botswana. J Appl.Sci Environ. Mgt June. 2006. Vol. 10 (2) 43-47. [2] Laverdire M and, Mateke S M Physical situation of indigenous fruit trees in Southern Africa Region. TBF, FAO Harare. [3] Nkhumeleni Jeoffrey Manenzhe, Natasha Potgieter, Teunis van Ree. Composition and antimicrobial activities of volatile components of Lippia javanica. Phytochemistry 65 (2004) 23332336. [4] Kwang-Geun Lee and Takayuki Shibamoto. Analysis of volatile components isolated from Hawaiian green coffee beans (Coffea arabica L.). Flavour Fragr. J. 2002; 17: 349351. [5] Clara E. Quijano, Guilhermo Salamanca and Jorge A. Pino. Aroma volatile constituents of colombian varieties of mango (Mangifera indica L). Flavour Fragr. J. 2007; 22: 401406. [6] Alasdair Sides, Kevin Robards, Stuart Helliwel. Development in extraction techniques and their application to analysis of volatiles in foods. Trend in analytical chemistry, vol. 19 nr.5, 2000. [7] R. Rawat, A. Gulati, G.D.Kiran Babu, R. Acharya, V.K. Kaul, B. Singh. Characterization of components of Kangara orthodox blacktea by gas chromatography-mass spectrometry. 2002

Presented at ICEF11 (May 22-26, 2011 Athens, Greece) as paper FMS058.