title: Simultaneous determination of adenine nucleotides, creatine phosphate and creatine in rat liver by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry.
abstract: A high performance liquid chromatography-electrospray ionization-tandem mass spectrometric method (HPLC-ESI-MS/MS) was developed for simultaneous determination of adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), adenosine 5'-monophosphate (AMP), creatine phosphate (CP), and creatine in rat liver. After extraction with pre-cooled (4 °C) methanol/water (1:1, v/v), the analytes were separated on a porous graphitic carbon (Hypercarb) column (2.1 mm × 150 mm, 5 μm) using a programmed gradient elution with a mobile phase consisting of 2 mmol/L ammonium acetate in water and 2 mmol/L ammonium acetate in acetonitrile (pH=10.0). The analytes were detected in a way of multiple reaction monitoring (MRM) under negative scan mode by a triple quadrupole mass spectrometer with electrospray ionization (ESI). An external calibration method with linear ranges from 10 to 5000 ng/mL for the five target compounds was used for quantification with a correlation coefficients≥0.9973. The limits of detection and limits of quantification for all analytes were in ranges from 0.50 to 1.5 ng/mL and 1.6 to 0.5 ng/mL, respectively. The average recoveries spiked in three levels were from 77.2% to 102% and precisions expressed in RSDs were from 0.2% to 4.8%. The established method was successfully applied to determination of ATP, ADP, AMP, CP and creatine in liver tissue.
authors.affiliation: West China School of Public Health, Sichuan University, No. 17, Section 3, South Renmin Road, Chengdu, Sichuan 610041, People's Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Jiang
authors.forename: Chengjun
authors.initials: C
authors.lastname: Sun
authors.forename: Xueqin
authors.initials: X
authors.lastname: Ding
authors.forename: Ding
authors.initials: D
authors.lastname: Yuan
authors.forename: Kefei
authors.initials: K
authors.lastname: Chen
authors.forename: Bo
authors.initials: B
authors.lastname: Gao
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen
authors.forename: Aimin
authors.initials: A
authors.lastname: Sun
mesh_terms: D000227:Adenine Nucleotides / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003401:Creatine / Q000032:analysis* / Q000378:metabolism; D057230:Limit of Detection; D008099:Liver / Q000378:metabolism; D010725:Phosphocreatine / Q000032:analysis* / Q000378:metabolism; D051381:Rats; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000227:Adenine Nucleotides; D010725:Phosphocreatine; D003401:Creatine