Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Optimized Formation of Benzyl Isothiocyanate by Endogenous Enzyme and its Extraction from Carica Papaya Seed

Wen-Zhao Li, Yi-Deng Du, Yan Zhang, Ying-Zhou Chi, Zong-Yi Shi, Wen Chen, Mei-Juan Ruan, Hua-Ping Zhu

Key Laboratory of Food Nutrition and Safety (Tianjin University of Science and Technology), Ministry of Education, Tianjin 300457, China;

For correspondence:- Hua-Ping Zhu Email: yideng126@126.com Tel:+8615122396920

Received: 29 April 2014 Accepted: 13 July 2014 Published: 18 August 2014

Citation: Li W, Du Y, Zhang Y, Chi Y, Shi Z, Chen W, et al. Optimized Formation of Benzyl Isothiocyanate by Endogenous Enzyme and its Extraction from Carica Papaya Seed. Trop J Pharm Res 2014; 13(8):1303-1311 doi: 10.4314/tjpr.v13i8.15

© 2014 The authors.
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Abstract

Purpose: To use endogenous myrosinase in Carica papaya seed to convert benzyl glucosinolate (BG) to benzyl isothiocyanate (BITC) and then extract it for further studies.

Methods: Process variables including seed powder particle size, sample-to-solvent ratio, pH of buffer solution, enzymolysis temperature, enzymolysis time were investigated using single-factor experiments and response surface methodology coupled with Box-Behnken design for further optimization of conversion conditions. The formed benzyl isothiocyanate was extracted by steam distillation method and purified by thin-layer chromatography (TLC). Relevant process variables were also studied. Finally, the purified benzyl isothiocyanate was analyzed by gas chromatography-mass spectrometer (GC-MS) and compared to BITC standard.

Results: Optimum enzymolysis conditions were seed powder particle size, 90 - 120 μm; sample-to-solvent ratio, 1:20; pH of buffer solution, 4.8; enzymolysis temperature, 40 0C; and enzymolysis time, 27 min. Enzymolysis liquid was steam-distilled for 2 h, extracted twice using dichloromethane (DCM) (using 3 times the volume of distillate each time) as extractant. Finally, 1:20 was adopted as the optimal dichloromethane-petroleum ether (DCM-PE) elution proportion to wash out the unnecessary compounds.

Conclusion: The result showed that benzyl isothiocyanate was formed under the optimized conditions and was no doubt the basic component of the extraction.

Keywords: Carica papaya, Benzyl isothiocyanate, Endogenous myrosinase, Enzymolysis, Extraction