Profiling and Preparation of Metabolites from Pyragrel in Human Urine by Online Solid-Phase Extraction Coupled with High Performance Liquid Chromatography Tandem Mass Spectrometry Followed by a Macroporous Resin-Based Purification Approach.
Pyragrel, a new anticoagulant drug, is derived from the molecular combination of ligustrazine and ferulic acid. Pyragrel showed significant inhibitory activity against platelet aggregation induced by adenosine diphosphate (ADP), and had been approved for a phase I clinical trial by CFDA. To characterize the metabolites of Pyragrel in human urine after intravenous administration, a reliable online solid-phase extraction couple with high performance liquid chromatography tandem mass spectrometry (online SPE-HPLC-MSn) method was conceived and applied. Five metabolites were detected and tentatively identified, which suggested that the major metabolic pathways of Pyragrel in human were double-bond reduction, double-bond oxidation, and then followed by glucuronide conjugation. Two main metabolites were then prepared using β-glucuronide hydrolysis and macroporous resin purification approach followed by preparative high-performance liquid chromatography (PHPLC) method, with their structures confirmed on the basis of nuclear magnetic resonance (NMR) data. This study provided information for the further study of the metabolism and excretion of Pyragrel.
Department of Pharmaceutical Analysis, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China. yoyo0132@163.com.
Xin
X
Zhao
Shanghai Key Laboratory for Pharmaceutical Metabolite Research, School of Pharmacy, Second Military Medical University, Shanghai 200433, China. jiangjingj8@163.com.
Jingjing
J
Jiang
Shanghai Key Laboratory for Pharmaceutical Metabolite Research, School of Pharmacy, Second Military Medical University, Shanghai 200433, China. diabloyg@126.com.
Guang
G
Yang
Center of Clinical Pharmacology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, China. cellahuang1988@163.com.
Jie
J
Huang
Center of Clinical Pharmacology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, China. ygp9880@126.com.
Guoping
G
Yang
Hefei Institute of Pharmaceutical Industry Co., Ltd., Wenshan Road, Hefei 230601, China. hgwhipi@hotmail.com.
Guangwei
G
He
Hefei Institute of Pharmaceutical Industry Co., Ltd., Wenshan Road, Hefei 230601, China. chuzhaoxing@163.com.
Zhaoxing
Z
Chu
Department of Pharmaceutical Analysis, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China. hangtj@cpu.edu.cn.
Taijun
T
Hang
Shanghai Key Laboratory for Pharmaceutical Metabolite Research, School of Pharmacy, Second Military Medical University, Shanghai 200433, China. hangtj@cpu.edu.cn.|Department of Clinical Pharmacy, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, No. 100 Haining Road, Shanghai 200080, China. hangtj@cpu.edu.cn.|Laboratory of Drug Metabolism & Pharmacokinetics, School of Medicine, Tongji University, No. 1239 Siping Road, Shanghai 200092, China. hangtj@cpu.edu.cn.
Guorong
G
Fan
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003373:Coumaric Acids / Q000737:chemistry*; D006801:Humans; D055432:Metabolomics / Q000379:methods*; D011719:Pyrazines / Q000737:chemistry*; D052616:Solid Phase Extraction / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D014556:Urine / Q000737:chemistry*
D003373:Coumaric Acids; D011719:Pyrazines; C004999:ferulic acid; C017953:tetramethylpyrazine
PHPLC; Pyragrel; human urine; macroporous resin; metabolite; online SPE-HPLC-MSn