ABSTRACT
Objective: To develop and validate an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC- MS/MS) method for the simultaneous determination of 11 components (calycosin, calycosin-7-glucoside, ginsenoside Rg1, ginsenoside Re, formononetin, astragaloside IV, ginsenoside Rf, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, and ginsenoside Rb1) in Kang'ai Injection (KI). Methods: Multiple reaction monitoring (MRM) with a polarity-switching electrospray ionization (ESI) source between positive and negative mode was used for the quantification. The 11 components were separated within 7.0 min on a Phenomenex Luna C18 column (100 mm × 3.0 mm, 2.6 μm) using a mobile phase consisted of methanol, acetonitrile, 5 mmol/L ammonium acetic-0.05 mmol/L sodium acetate water solution with gradient elution. Results: The linear relationships showed well in the area of determination. The average recovery varied between 95.2% and 104.4% with RSD ≤ 4.64%. The precision, repeatability, and stability of the method were good for the determination of 11 components. The content ranges of calycosin, calycosin-7-glucoside, ginsenoside Rg1, ginsenoside Re, formononetin, astragaloside IV, ginsenoside Rf, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, and ginsenoside Rb1 in four batches of KI were 0.004-0.006 μg/mL, 0.002-0.003 μg/mL, 125.75-148.00 μg/mL, 51.75-77.00 μg/mL, 0.010-0.013 μg/mL, 51.50-87.75 μg/mL, 27.83-30.73 μg/mL, 4.23-5.15 μg/mL, 8.40-13.35 μg/mL, 17.33-27.68 μg/mL, and 9.03-11.00 μg/mL, respectively. Conclusion: A new rapid and reliable UPLC-MS/MS method was developed and validated to evaluate the quality of KI for the first time.