SIMULTANEOUS QUANTIFICATION OF THREE NAPHTHOQUINONES FROM IMPATIENS BALSAMINA L. LEAVES USING VALIDATED RP-HPLC METHOD

Impatiens balsamina belonging to family Balsaminaceae is commonly known as terda. The plant has an extensive range of phyto-constituents like naphthoquinones, coumarins, glycosides, phenolic acids, flavonoids, anthocyanidins and steroids which have effective antimicrobial, anti-anaphylaxis, anti-allergic, antipruritic, and anti-inflammatory activity. Potent naphthoquinones like lawsone, methylene-3, 3’-bilawsone and lawsone methyl ether are present in the leaves of the plant that exhibits was proved to possess an intensive antitumor activity. Taking into account these ethnobotanical, nutritional, medicinal and commercial values of I. balsamina, a simple, rapid and sensitive High-Performance Liquid Chromatography method for quantification of three naphthoquinones in is developed. Jasco CrestPak C8 (4.6×250mm, 5μm) RP-HPLC column was used as the stationary phase. Sodium acetate (pH3.2): ACN (50:50% v/v) was used as mobile phase with flow rate 1ml/min and detection at a wavelength of 280nm using PDA multi-wavelength Detector. The developed method was validated for linearity, accuracy, precision, and system suitability. LOD for naphthoquinones was found to be 200ng/mL, whereas LOQ for the same was found to be 600ng/mL. The method was found to be linear over the range 5 μg/mL to 15 μg/mL with the coefficient of regression 0.997, 0.990 and 0.998 for lawsone, methylene-3, 3’-bilawsone and lawsone methyl ether respectively. The result indicated that the developed HPLC method could be successfully used for quantitative estimation of naphthoquinones from Impatiens balsamina.