Microwave Assisted Extraction of Berberine and Preparation of Berberine Hydrochloride from Berberis Aristata Variety of Nepal, and Quantification using RP-HPLC and HPTLC Methods

Abstract

Berberis aristata a Himalayan woody spiny shrub with yellow flowers and red berries commonly called as Daruharidra in Sanskrit and locally in Nepal is called as Chutro or Chitra. The root and stem are the two parts widely used in traditional medicines of India and China. Berberine is the key active ingredient present in stem and root parts. Berberine hydrochloride is the derivative of berberine.  The present study aimed to study the microwave assisted extraction of berberine and its conversion into berberine hydrochloride and quantifying by RP-HPLC and HPTLC methods.In the present paper we have mentioned microwave assisted extraction of berberine and preparation of berberine hydrochloride in detail. Berberine extracted from roots of Berberis aristata using microwave assisted extraction in 80% ethanol to obtain 20% pure berberine crude by HPTLC densitometry at 350 nm absorption. The crude berberine was further purified to berberine hydrochloride by adding 10% Hcl in aqueous solution of berberine and allowed to crystallize at 5 0C over 24 hours. The crystals were further purified and recrystallized in ethanol and subjected to RP-HPLC. Reverse phase HPLC was carried out on Shimadzu UV detector at wave length of 265 nm using Acetonitrile-0.1% phosphoric acid solution (50:50) (add 0.1g sodium dodecyl sulfonate per 100ml) as the mobile phase; Phenomenex RP-column (250 mm × 4.6 mm, 5 μm), With flow rate 1.0 mL/minute, 5 µL injection volume, column temperature 25 0C for run time of 35 minutes, and retention time of berberine hydrochloride was 12.008 minutes with purity of 82%  and recovery of 90% yield obtained.