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Advanced Pharmaceutical Bulletin
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Article History
Submitted: 12 May 2015
Revised: 24 Aug 2015
Accepted: 25 Aug 2015
First published online: 30 Nov 2015

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Adv Pharm Bull. 2015;5(4):587-591 doi: 10.15171/apb.2015.079
PMID:26819933        PMCID:PMC4729348

Determination of Four Major Saponins in Skin and Endosperm of Seeds of Horse Chestnut (Aesculus Hippocastanum L.) Using High Performance Liquid Chromatography with Positive Confirmation by Thin Layer Chromatography

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Zead Helmi Mahmoud Abudayeh 1 * , Khaldun Mohammad Al Azzam 2 * , Ahmad Naddaf 1, Uliana Vladimirovna Karpiuk 3, Viktoria Sergeevna Kislichenko 4

1 Faculty of Pharmacy, Isra University, 11622 Amman, Jordan.
2 Department of Pharmaceutical Chemistry, Pharmacy Program, Batterjee Medical College for Sciences and Technology (BMC), 21442 Jeddah, Kingdom of Saudi Arabia.
3 Department of Pharmacognosy and Botany, National Medical University is the Name of O.O.Bogomolets, Ukraine.
4 National University of Pharmacy, Kharkiv, Ukraine.



Abstract
urpose: To separate and quantify four major saponins in the extracts of the skin and the endosperm of seeds of horse chestnut (Aesculus hippocastanum L.) using ultrasonic solvent extraction followed by a high performance liquid chromatography-diode array detector (HPLC-DAD) with positive confirmation by thin layer chromatography (TLC). Methods: The saponins: escin Ia, escin Ib, isoescin Ia and isoescin Ib were extracted using ultrasonic extraction method. The optimized extraction conditions were: 70% methanol as extraction solvent, 80 C as extraction temperature, and the extraction time was achieved in 4 hours. The HPLC conditions used: Zorbax SB-ODS-(150 mm × 2.1 mm, 3 m) column, acetonitrile and 0.10% phosphoric acid solution (39:61 v/v) as mobile phase, flow rate was 0.5 mL min-1 at 210 nm and 230 nm detection. The injection volume was 10 L, and the separation was carried out isothermally at 30 C in a heated chamber. Results: The results indicated that the developed HPLC method is simple, sensitive and reliable. Moreover, the content of escins in seeds decreased by more than 30% in endosperm and by more than 40% in skin upon storage for two years. Conclusion: This assay can be readily utilized as a quality control method for horse chestnut and other related medicinal plants.





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