ABSTRACT
Objective: To establish a high performance liquid chromatography/mass spectrometry (HPLC-MS/MS) for the simultaneous determination of the concentration in plasma of chlorogenic acid, new chlorogenic acid, cryptochlorogenic acid, gardenia component geniposide, shanzhiside, and genipin gentian diglucoside from Lonicerae Flos in Reduning Injection iv dripped in healthy human body. Methods: Sixteen healthy subjects were iv infused of Reduning Injection with 2 ampoules, a total of 20 mL diluted with 5% glucose injection to 250 mL, iv injection time was set for 90 min, venous blood was measured at the different time points before, during, and after infusion, respectively. The conditions of determination for chlorogenic acid and other ingredients were as follows: Inertsil ODS-2 chromatographic column (150 mm × 2.1 mm, 5 μm), mobile phase (acetonitrile, 0.1% formic acid), rolling gas 137.9 kPa (20 psi), collision gas 56.16 kPa (8 psi), spray voltage -4 500 V. The LC-MS/MS conditions for determination of shanzhiside, geniposide and genipin gentian diglycoside were as follows: Ecosil C18 (150 mm × 4.6 mm, 5 μm), the mobile phase (methanol, 20 mmol/L ammonium formate, 0.1% formic acid, and 10% methanol), rolling gas 96.53 kPa (14 psi), collision gas 56.16 kPa (8 psi), spray voltage 4 500 V. Results: The tmax of neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid were 1.31, 1.38, and 144 h; Cmax were 1 241, 329 4, and 2 121 ng/mL; AUC0~t were 1 972, 5 351, and 3 596 ng·h/mL; MRT0~t were 0.708, 0.790, and 0.899 h; CL were 10.3, 385, and 5.73 L/h; Vd were 16.7, 766, and 10.7 L; t1/2 were 1.13, 1.36, and 1.27 h, respectively. The tmax of geniposide, shanzhiside, and genipin gentian diglucoside were 1.41, 1.47, and 1.47 h; Cmax were 4.49, 0.288, and 0.541 μg/mL; AUC0~t were 7.41, 0.671, and 1.22 μg·h/mL; MRT0~t were 0.856, 1.59, and 152 h; CL were 2.74, 30.0, and 16.6 L/h; Vd were 5.63, 60.9, and 35.2 L; t1/2 were 1.42, 1.42, and 1.47 h. Conclusion: The method is simple, accurate, and can be used for in vivo pharmacokinetic study on Reduning Injection.