title: 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.
abstract: OBJECTIVE
  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 μm) 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 μL, 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.
  
  
  CONCLUSIONS
  This assay can be readily utilized as a quality control method for horse chestnut and other related medicinal plants.
authors.affiliation: Faculty of Pharmacy, Isra University, 11622 Amman, Jordan.
authors.forename: Zead Helmi Mahmoud
authors.initials: ZH
authors.lastname: Abudayeh
authors.affiliation: Department of Pharmaceutical Chemistry, Pharmacy Program, Batterjee Medical College for Sciences and Technology (BMC), 21442 Jeddah, Kingdom of Saudi Arabia.
authors.forename: Khaldun Mohammad
authors.initials: KM
authors.lastname: Al Azzam
authors.affiliation: Faculty of Pharmacy, Isra University, 11622 Amman, Jordan.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Naddaf
authors.affiliation: Department of Pharmacognosy and Botany, National Medical University is the Name of O.O.Bogomolets, Ukraine.
authors.forename: Uliana Vladimirovna
authors.initials: UV
authors.lastname: Karpiuk
authors.affiliation: National University of Pharmacy, Kharkiv, Ukraine.
authors.forename: Viktoria Sergeevna
authors.initials: VS
authors.lastname: Kislichenko
keywords: Aesculus hippocastanum L.; HPLC; Horse chestnut; Saponins; TLC; Ultrasonic extraction
references.citation: Mutat Res. 2005 Jul 1;574(1-2):124-38
references.pmid: 15914212
references.citation: Phytochem Anal. 2013 Nov-Dec;24(6):513-9
references.pmid: 23508832
references.citation: Molecules. 2014 Sep 15;19(9):14625-36
references.pmid: 25225723
references.citation: Acta Pol Pharm. 2013 May-Jun;70(3):517-22
references.pmid: 23757942
references.citation: J Ethnopharmacol. 2005 Aug 22;100(1-2):131-4
references.pmid: 16006081
references.citation: Ultrason Sonochem. 2011 Jan;18(1):243-9
references.pmid: 20637676
references.citation: Plant Foods Hum Nutr. 2005 Mar;60(1):25-9
references.pmid: 15898356
references.citation: Chem Pharm Bull (Tokyo). 1994 Jun;42(6):1357-9
references.pmid: 8069982
references.citation: Ultrason Sonochem. 2013 Jul;20(4):1026-32
references.pmid: 23414834
references.citation: Acta Pol Pharm. 2011 May-Jun;68(3):403-8
references.pmid: 21648195