Adv Pharm Bull. 2015;5(4):587-591.
doi: 10.15171/apb.2015.079
PMID: 26819933
PMCID: PMC4729348
Scopus id: 84949653006
  Abstract View: 472
  PDF Download: 188

Short Communication

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

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.


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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Submitted: 12 May 2015
Revised: 24 Aug 2015
Accepted: 25 Aug 2015
First published online: 30 Nov 2015
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