Rapid Identification and Simultaneous Quantification of Multiple Constituents in Nao-Shuan-Tong Capsule by Ultra-Fast Liquid Chromatography/Diode-Array Detector/Quadrupole Time-of-Flight Tandem Mass Spectrometry.

Abstract

A rapid and high-sensitive ultra-fast liquid chromatography coupled with a diode-array detector and a quadrupole/time-of-flight mass spectrometry (MS) method was established and validated for the chemical profiling of Nao-shuan-tong capsule (NSTC) and simultaneous quantification of five major constituents. A total of 59 components including monoterpene glycosides, flavonoids, sesquiterpenoids, ketosteroids, thiophenes, organic acids and alkaloids were identified or tentatively characterized in NSTC based on the accurate mass and tandem MS behavior. Five major bioactive constituents were chosen as the chemical indexes of holistic quality evaluation and quantified simultaneously. All calibration curves showed good linear regression (r(2) > 0.9991) in the range 25.2-510, 145-2,900, 1.84-36.8, 2.61-52.2 and 3.25-26.2 μg/mL for gastrodin, paeoniflorin, typhaneoside, β-ecdysterone and isorhamnetin-3-O-neohesperidoside, respectively. It also showed good precision, stability and accuracy for quantification of these five compounds. The limit of detections and limit of quantitations for the analytes ranged from 0.14 to 1.09 μg/mL and from 0.47 to 3.63 μg/mL, respectively. The validated quantification method was applied to analyze 10 batches of commercial NSTC. These results will provide a basis for quality control of the production process and the further pharmacological study in vivo of NSTC.