title: Measurement of hydroxysafflor yellow A in human urine by liquid chromatography-tandem mass spectrometry.
abstract: A rapid and specific high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the quantification of hydroxysafflor yellow A (HSYA) in human urine with isorhamnetin-3-O-neohespeidoside as internal standard (IS). HSYA and IS were extracted from urine samples by simple solid-phase extraction and separated on an Agilent Zorbax SB C18 column (4.6 mm × 150 mm, 5 μm) with the mobile phase of 0.2 mM ammonium acetate: methanol (30/70, v/v) at a flow rate of 0.4 mL/min. Polar endogenous interferences eluted in 0.1-2.5 min were switched into waste channel by the Valve Valco, to reduce the possible matrix effect for MS detection in each run. The MS detection of analytes was performed on a tandem mass spectrometer equipped with an electrospray ionization source in negative mode using multiple-reaction monitoring. The MS/MS ion transitions monitored were m/z 611.3→491.2 for HSYA and m/z 623.2→299.2 for IS. The method was fully validated for selectivity, sensitivity, linearity, precision, accuracy, recovery, matrix effect and stability, and then was applied to the urinary excretion study of injectable powder of pure HSYA in healthy Chinese volunteers for the first time. The results suggested that urine was the main excretion way of HSYA in healthy volunteers, further demonstrating the feasibility and necessity of our current method.
authors.affiliation: Department of Clinical Pharmacology, Affiliated Hospital of Nanjing University of Chinese Medicine, No. 155 Hanzhong Road, Nanjing 210029, China. Electronic address: changyinli2008@126.com.
authors.forename: Chang-Yin
authors.initials: CY
authors.lastname: Li
authors.affiliation: Department of Clinical Pharmacology, Affiliated Hospital of Nanjing University of Chinese Medicine, No. 155 Hanzhong Road, Nanjing 210029, China.
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Chu
authors.affiliation: Department of Clinical Pharmacology, Affiliated Hospital of Nanjing University of Chinese Medicine, No. 155 Hanzhong Road, Nanjing 210029, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Clinical Pharmacology, Affiliated Hospital of Nanjing University of Chinese Medicine, No. 155 Hanzhong Road, Nanjing 210029, China.
authors.forename: Bing-Ting
authors.initials: BT
authors.lastname: Sun
authors.affiliation: Department of Clinical Pharmacology, Affiliated Hospital of Nanjing University of Chinese Medicine, No. 155 Hanzhong Road, Nanjing 210029, China.
authors.forename: Guo-Liang
authors.initials: GL
authors.lastname: Dai
authors.affiliation: Department of Clinical Pharmacology, Affiliated Hospital of Nanjing University of Chinese Medicine, No. 155 Hanzhong Road, Nanjing 210029, China.
authors.forename: Shi-Jia
authors.initials: SJ
authors.lastname: Liu
authors.affiliation: Department of Clinical Pharmacology, Affiliated Hospital of Nanjing University of Chinese Medicine, No. 155 Hanzhong Road, Nanjing 210029, China. Electronic address: wzhju333@163.com.
authors.forename: Wen-Zheng
authors.initials: WZ
authors.lastname: Ju
mesh_terms: D000328:Adult; D002599:Chalcone / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008297:Male; D011809:Quinones / Q000302:isolation & purification / Q000652:urine*; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D011809:Quinones; C085278:hydroxysafflor yellow A; D002599:Chalcone
keywords: Column switching; Excretion; Human urine; Hydroxysafflor yellow A; LC–MS/MS; SPE