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Previous studies have reported that a number of organic compounds are present in kaffir lime (Citrus hystrix DC.) leaf extracts. Further research is needed to purify these compounds and determine which are biologically active. The objective of this study is to identify the volatile organic compounds of kaffir lime leaf crude extracts and fractions and to study their bioactivity. Fractionation was performed by the double maceration method, using hexane as the second solvent. TLC was performed to analyze the qualitative separation, whereas the individual constituents were detected using GC-MS. Our results showed that chloroform and ethyl acetate crude extracts contained various volatile organic compounds such as fatty acids, fatty alcohols, prenol lipids, sterol lipids, terpenoids and long chain alkanes. Fractionation separated these compounds into non-hexane fractions, which contained less volatile compounds, and hexane fractions. The volatile compounds of non-hexane fractions were identified to be long chain alkanes, meanwhile the hexane fractions contained terpenoids, fatty acids, fatty alcohols, prenol lipids and sterol lipids. Palmitic acid and terpenoids, such as citronellyl propionate, nerolidol, citronella and caryophyllene oxide were found to be the most dominant bioactive compounds in chloroform and ethyl acetate crude extract and their hexane fractions, which were reported to possess cytotoxicity against cancer cells. Meanwhile in non-hexane fractions, long chain alkanes such as triacontane and hentriacontane were found to be the most dominant bioactive compound which also possessed cytotoxic effect. In conclusion, fractionation using the double maceration method yielded different volatile organic compounds composition with different biological activities. The crude extracts and fractions of kaffir lime leaves were potential to be developed as a traditional medicine for cancer treatment.

Volatile Organic Compounds of Kaffir Lime (Citrus hystrix DC.) Leaves Fractions and their Potency as Traditional Medicine