Simple and rapid determination of adenosine in human synovial fluid with high performance liquid chromatography-mass spectrometry.
A simple, fast, sensitive and selective reversed-phase high performance liquid chromatography-mass spectrometry coupling with an electrospray ionization (ESI) interface method is described for the determination of adenosine in human synovial fluid. This method involved the use of the [M + H](+)ions of adenosine and 2-chloroadenosine (internal standard for the assay) at m/z 268 and 302 in positive ion mode with selective ion monitoring (SIM). Separation was carried out on a 2.0 x 150 mm Shimadzu VP-ODS column by using an isocratic elution with a mobile phase consisting of water (94%),methanol (5%) and formic acid (1%). No interference with the components of the biological matrix was observed in the determination conditions. The calibration curve was linear in the range of 0.2-140 microgml(-1). The limits of quantification (LOQ) and detection (LOD) were 0.2 and 0.03 microgml(-1), respectively. The standard recoveries were between 93.3 and 104.0%. The method was successfully applied to determination of adenosine in some synovial fluids of patients affected by rheumatoid arthritis.
Huang
Lan-Fang
LF
College of Chemistry and Chemical Engineering of Central South University, Changsha 410083, P. R. China. fl18huang@yahoo.com.cn
Guo
Fang-Qiu
FQ
Liang
Yi-Zeng
YZ
Li
Bo-Yang
BY
Cheng
Ben-Mei
BM
D000241:Adenosine / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013582:Synovial Fluid / Q000737:chemistry*
D000241:Adenosine