Development of Modifed Extract and HPLC Analytical Method for Determination of Rutin in Modifed Extract of Longan seed

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Chutipan Nusuk
Chamnan Patarapanich
Bodin Tuesuwan


Introduction: Several  studies  of  chemicals  in  longan  (Dimocarpuslongan  Lour.)  seed  have  been suggested  to have positive effect on defcit  in  learning and memory.  In our study,  the ethyl acetate extracts of longan seeds found the positive effect on learning and memory impairment after bilateral carotid arteries (2-VO) occlusion in mice. Materials and Methods: The Fractionation of the ethyl acetate part by conventional chromato-graphic technique together with the screening activities was performed. A HPLC-analytical method was developed for determination of rutin, a chemical marker in modifed extract using reverse phase column with isocratic elution. Hesperidin was used as an internal standard. Results: We obtained pure four major compounds with physical and spectroscopic data acquired indicated as quercetin, gallic acid, propyl gallate and rutin. Rutin showed to be a one of the most active principle compounds at dose 100mg/kg; we proposed rutin as chemical marker for the modifed herbal extract. The method was validated according to ICH guideline and showed linearity range of 0.31-1.54µg/ml of rutin (r2=0.9940). The percentage recovery (%R) was in the range of 99.30-101.98 and the relative standard deviation (%RSD) was not more than 2. The limit of detection and limit of quantifcation of rutin were 0.13 and 0.38 µg/ml, respectively. Conclusion: We prepared relatively stable herbal extract from longan seeds by removing the unstable phenolic portion for further chemical and preclinical studied. Also, High-performance liquid chromatogra-phy technique was used for quality control of modifed extract of longan seed. Proper validation of HPLC method according to ICH guidelines.

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