Simultaneous determination of seven hydrophilic bioactive compounds in water extract of Polygonum multiflorum using pressurized liquid extraction and short-end injection micellar electrokinetic chromatography

Simultaneous determination of seven hydrophilic bioactive compounds in water extract of Polygonum multiflorum using pressurized liquid extraction and short-end injection micellar electrokinetic chromatography

Ka-meng Lao , Dong-qi Han , Xiao-jia Chen , Jing Zhao, Tie-jie Wang and Shao-ping Li

Abstract

Background: Polygoni Multiflori Radix, He-Shou-Wu in Chinese, is a widely used traditional Chinese medicine.Clinically, water decoction is the major application form of He-Shou-Wu. Therefore, simultaneous determination of bioactive compounds in water extract is very important for its quality control.

Results: A pressurized liquid extraction and short-end injection micellar electrokinetic chromatography (MEKC) were first developed for simultaneous determination of seven hydrophilic bioactive compounds in water extract of He-Shou-Wu. The influence of parameters, such as pH, concentration of phosphate, SDS and HP-β-CD, capillary temperature and applied voltage, on the analysis were carefully investigated. Optimum separation was obtained within 14 min by using 50 mM phosphate buffer containing 90 mM SDS and 2% (m/v) HP-β-CD (pH 2.5) at 15 kV and 20°C. All calibration curves showed good linearity (R2 >0.9978) within test ranges. The overall LOD and LOQ were lower than 2.0 μg/mL and 5.5 μg/mL, respectively. The RSDs for intra- and inter-day of seven analytes were less than 3.2% and 4.6%, and the recoveries were 97.0%-104.2%.

Conclusion: The validated method was successfully applied to the analysis of He-Shou-Wu samples, which is helpful for its quality control.

Proanthocyanidin B1 (>95%) and proanthocyanidin B2 (>95%) were purchased from Chengdu Biopurify Phytochemicals Co. Ltd (Chengdu, China).