Lekhooa Makhotso, Walubo Andrew, Du Plessis Jan J B, Matsabisa Motlalepula C, Molefe Duduzile
Department of Pharmacology, University of the Free State, Francie van Ziyl Drive, Tygerberg 7505, Parow Valley, Cape Town, South Africa.
Afr J Tradit Complement Altern Med. 2012 Feb 13;9(3 Suppl):27-39. doi: 10.4314/ajtcam.v9i3s.5. eCollection 2012.
PHELA is a herbal mixture of four African traditional medicinal plants that has been used for decades in wasting conditions and is now being developed by the Medical Research Council (MRC) as an immune booster for patients with compromised immune system. A chromatographic fingerprint of PHELA was needed for quality control purposes. Here, a comprehensive method for fingerprinting of PHELA using different chromatographic techniques is described. It involved extraction of the PHELA by either acidic or a simple 'salting-out' method, followed by Thin Layer Chromatography (TLC) analysis and/or preparative Column Chromatography (CC). The products were thereafter analyzed by High Performance Liquid Chromatography with UV-detector (HPLC-UV), HPLC with fluorescence-detector (HPLC-FL) and Gas-Chromatography with a Mass Selective Detector spectrometer (GC-MSD). The fingerprints were successfully used to differentiate PHELA from another common herbal product made from Hypericum perforatum (St. John's Wort), thereby illustrating its high potential for use in fingerprinting of PHELA and in differentiating it from other herbal medicines. By validating the different chromatographic techniques on the standardized extraction methods, this approach will enable wide application in quality control of PHELA using acceptable procedures, thereby promoting effective monitoring of the finished product in all countries where it will be used.
PHELA是一种由四种非洲传统药用植物组成的草药混合物,数十年来一直用于治疗消瘦病症,目前医学研究理事会(MRC)正在将其开发为免疫系统受损患者的免疫增强剂。为了进行质量控制,需要PHELA的色谱指纹图谱。本文描述了一种使用不同色谱技术对PHELA进行指纹图谱分析的综合方法。该方法包括通过酸性或简单的“盐析”方法提取PHELA,然后进行薄层色谱(TLC)分析和/或制备柱色谱(CC)。之后,使用带紫外检测器的高效液相色谱(HPLC-UV)、带荧光检测器的高效液相色谱(HPLC-FL)和带质量选择检测器光谱仪的气相色谱(GC-MSD)对产物进行分析。这些指纹图谱成功地用于区分PHELA与另一种由贯叶连翘(圣约翰草)制成的常见草药产品,从而表明其在PHELA指纹图谱分析以及将其与其他草药区分开来方面具有很高的潜力。通过在标准化提取方法上验证不同的色谱技术,这种方法将能够使用可接受的程序在PHELA的质量控制中广泛应用,从而促进在所有使用该产品的国家对成品进行有效监测。