Determination Of Flavonoid In Three Chinese Medicines By Liquid Chromatography-Tandem Mass Spectrometry

This topic combined highly sensitive mass spectrometry detection technology with high-performance high performance liquid chromatography technology in an manner, and established common detection methods of flavonoids in three Chinese Medicines.This paper mainly discussed mass spectrometry and chromatographic analysis conditions. the effect of molecular ion peak, capillary voltage, fragments ion peak and collision energy, moreover the effect of stationary phase,mobile phase, flow rate and column temperature were also investigated in detail.Under the optimized mass spectrometry and chromatography analysis conditions, HPLC-MS methods for the determination of the root of kudzu vine, Fructus Psoraleae, Scutellaria baicalensis were established successfully Linear range, precision, detection limit and accuracy were researched one by one.A liquid chromatography-tandam mass spectrometric (LC-MS/MS) method was developed for determination of Puerarin and Daidzein in Puerarin root. The separation was performed on a waters Xterra MS C18(3.9×150mm,5μm)column by using 50 mmol·L-1 formic acid solution and acetonitrile as mobile phase. The experiments were performed by electrospray ionization-tandem mass spectrometry in positive mode, and quantified by matrix-matched external standard method. The calibration curves showed good linearity in the concentration range of 50-500μg·L-1 and 50-50mg·L-1 with the better correlation coefficients(≥0.9991). The limits of detection was 5μg·L-1 for puerarin and daidzein. The mean recoveries were 96.6% and 97.4% repectively. The relative standard deviations were 2.3% and 3.1%(c=0.5mg-L-',n=6) for both flavonoid compounds. The method has been proved to be satisfactory in real simple determination.A liquid chromatography-tandam mass spectrometric (LC-MS/MS) method was developed for determination of bavachin and neobavaisoflavone in Malaytea Scurfpea Fruit. The separation was performed on a pheenomenex luna 5u Cg(150×4.60mm,5micron) column by using the elution with 10 mmol·L-1 ammonium acetate solution and acetonitrile. The residues were determined by electrospray ionization-tandem mass spectrometry in positive mode, quantified by matrix-matched external standard method. The calibration curves showed good linearity in 10μg·L-1-100mg·L-1 levels with the correlation coefficients of 0.9991 and 0.9994 respectively. The limits of detection were 7.8μg·L-1 and 5.7μg·L-1,The recoveries of samples at 0.1,1 and 10 mg·L-1 were 99.4% and 100.6%. The relative standard deviation were 1.3% and 2.1%(c=1mg·L-1,n=6). The method has been proved to be satisfactory in real simple determination.A liquid chromatography-tandam mass spectrometric (LC-MS/MS) method was developed for determination of Baicalin and Baicalein in Scutellaria baicalensis. The separation was performed on a Varian pursuit C18(100×2.0mm,3μm) column by using the elution with 0.1% formic acid solution and acetonitrile. The residues were determined by electrospray ionization-tandem mass spectrometry in positive mode, quantified by matrix-matched external standard method. The calibration curves showed good linearity in 50μg·L-1-50mg·L-1 levels with the correlation coefficients of 0.9994 and 0.9992 respectively. The limits of detection were 3.1μg·L-1 and 4.7μg·L-1.The recoveries of samples at 0.1,1 and 10 mg·L-1 were 109.6% and 94.1%. The relative standard deviation were 1.1% and 1.9%(c=1mg·L-1,n=6). The method has been proved to be satisfactory in real simple determination.