WHO strategy on traditional medicine is based on security and quality of phytomedicine. Commonly adulterated affecting mostly metabolic and sexual dysfunction drugs. Control quality of those phytomedicines requires development of strategy and techniques applicable to them. Among the techniques, Reverse Phase Liquid Chromatography is the most used and has been developed in these studies to assess a protocol to characterize Gardenia aqualla leaves extract. The method consists in determining chromatographic conditions using organic and pH gradient models based on water and acetonitrile combined with pH modifiers made up of formic acid (AF) and ammonium hydroxide (NH3). Results show that extracts contain mainly acidic compounds quickly eluted by NH3 and more retained by AF. Optimal pH range for separation is 3 - 7 corresponding to 1.59 mM of NH3 and 6.55 mM of AF. In these conditions, elution of many polar compounds could be effective using a C18 based-deactivated column in a short period of time.
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