Establishment of a Method for Determination of Serum Adenosine and Inosine by Chromatography Tandem Mass Spectrometry
Objective To establish a high-performance liquid chromatography-tandem massspectrometry(LC-MS)method for the determination of serum adenosine and inosine concentrations,and assess the stability of adenosine and inosine in serum samples.Methods The quantification of serum adenosine and inosine was performed using positive ion mode and multiple reaction monitoring(MRM)scanning mode,with lamivudine serving as an internal standard.The ion pairs used for quantitative analysis were m/z:268.1/136.1,m/z:268.9/137.1 and m/z:230.0/112.0 for adenosine,inosine and internal standard,respectively.The stability of serum adenosine and inosine in 4 ℃ and-80 ℃ were investigated.Results The established method in this study enabled simultaneous quantification of serum adenosine and inosine concentrations.The detection limit for adenosine and inosine were 2.5-1 000 μg/L(R2＞0.99)and 2.5-1 000 μg/L(R2＞0.99),respectively.The accuracy of adenosine and inosine detection ranged from 85.77％to 97.57％,with a relative deviation of precision less than 8％.Upon storage at 4 ℃ for 4 h,adenosine and inosine degraded to 3.33％and 88.30％of the detected concentration at 1 h,respectively.After storing at-80 ℃ for 24 h,adenosine and inosine degraded to 71.70％and 84.00％of the detected concentration at 1 h,respectively.The levels of serum adenosine and inosine in 29 healthy subjects were detected by the constructed method.The results showed that the serum adenosine and inosine concentrations in healthy subjects were(452.471±293.793)μg/L and(30.547±14.160)μg/L respectively,with no significant difference between male and female(P＞0.050).Conclusion The method established in this study allows accurate and precise simultaneous quantification of adenosine and inosine concentrations in serum samples.Adenosine was found to be susceptible to degradation,highlighting the suitability of inosine to serve as a more stable biomarker for future diagnostic applications.
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