Simultaneous determination of creatine phosphate, creatine and 12 nucleotides in rat heart by LC-MS/MS.
A simple, rapid and sensitive LC-MS/MS method was developed and validated for simultaneous determination of creatine phosphate (CP), creatine (Cr) and 12 nucleotides in rat heart. The analytes, ATP, ADP, AMP, GTP, GDP, GMP, CTP, CDP, CMP, UTP, UDP, UMP, CP, Cr, were extracted from heart tissue with pre-cooled (0°C) methanol/water (1:1, v/v) and separated on a Hypersil Gold AQ C18 column (150mm×4.6mm, 3μm) using an isocratic elution with a mobile phase consisting of 2mmol/L ammonium acetate in water (pH 10.0, adjusted with ammonia). The detection was performed by negative ion electrospray ionization in selective reaction monitoring mode (SRM). In the assay, all the analytes showed good linearity over the investigated concentration range (r>0.99). The accuracy was between 80.7% and 120.6% and the precision expressed in RSD was less than 15.6%. This method was successfully applied to measure the concentrations of the 12 nucleotides, creatine phosphate and creatine in rat heart for the first time.
Department of Pharmacology and Toxicology, Tasly R&D Institute, Tianjin Tasly Group Co., Ltd, Tianjin 300410, China; Heilongjiang University of Traditional Chinese Medicine, Harbin 150040, China.
Jun-mei
JM
Wang
Department of Pharmacology and Toxicology, Tasly R&D Institute, Tianjin Tasly Group Co., Ltd, Tianjin 300410, China.
Yang
Y
Chu
Department of Pharmacology and Toxicology, Tasly R&D Institute, Tianjin Tasly Group Co., Ltd, Tianjin 300410, China. Electronic address: Liwei_wzhy@163.com.
Wei
W
Li
Department of Pharmacology and Toxicology, Tasly R&D Institute, Tianjin Tasly Group Co., Ltd, Tianjin 300410, China.
Xiang-yang
XY
Wang
Department of Pharmacology and Toxicology, Tasly R&D Institute, Tianjin Tasly Group Co., Ltd, Tianjin 300410, China.
Jia-hua
JH
Guo
Department of Pharmacology and Toxicology, Tasly R&D Institute, Tianjin Tasly Group Co., Ltd, Tianjin 300410, China.
Lu-lu
LL
Yan
Department of Pharmacology and Toxicology, Tasly R&D Institute, Tianjin Tasly Group Co., Ltd, Tianjin 300410, China.
Xiao-hui
XH
Ma
Heilongjiang University of Traditional Chinese Medicine, Harbin 150040, China.
Ying-li
YL
Ma
Department of Pharmacology and Toxicology, Tasly R&D Institute, Tianjin Tasly Group Co., Ltd, Tianjin 300410, China; Heilongjiang University of Traditional Chinese Medicine, Harbin 150040, China.
Qi-hui
QH
Yin
Tianjin State Key Laboratory of Pharmacokinetics and Pharmacodynamics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China.
Chang-xiao
CX
Liu
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003401:Creatine / Q000032:analysis*; D057230:Limit of Detection; D008297:Male; D009206:Myocardium / Q000737:chemistry*; D009711:Nucleotides / Q000032:analysis*; D010725:Phosphocreatine / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D053719:Tandem Mass Spectrometry / Q000379:methods*
D009711:Nucleotides; D010725:Phosphocreatine; D003401:Creatine
Creatine; Creatine phosphate; LC–MS/MS; Nucleotides; Rat heart