JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE) ›› 2021, Vol. 41 ›› Issue (3): 400-405.doi: 10.3969/j.issn.1674-8115.2021.03.021

• Techniques and methods • Previous Articles     Next Articles

Determination of inosine in human plasma by high performance liquid chromatography-tandem mass spectrometry combined with solid-phase extraction

Guan-qin JIN(), Li SUN(), Ling-hong XIA, Hou-wen LIN   

  1. Department of Pharmacy, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
  • Received:2020-04-20 Online:2021-03-28 Published:2021-04-06
  • Contact: Li SUN;
  • Supported by:
    Scientific Research Project of Science and Technology Commission of Shanghai Municipality(1741196400)

Abstract: Objective

·To establish a solid-phase extraction-high performance liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS) method for the determination of inosine in human plasma.


·Inosine and internal standard in human plasma were quantitatively analyzed with electrospray ionization positive ion mode and multiple reaction monitoring scan analysis. The ion pairs used for quantitative analysis were m/z 269.1/137.1 (inosine) and m/z 230.2/112.2 (internal standard). The results of method validation included limit of quantification, accuracy, intra-day and inter-day precision, extraction recovery, and stability tests in different conditions.


·The results of method validation showed that the calibration curve of inosine in human plasma exhibited a good linearity (R2>0.999) over the range from 28.5 to 912.0 ng/mL, and quantitative lower limit was 28.5 ng/mL. Accuracy was 96.9%?103.8%, and the relative standard deviations of intra-day and inter-day precision were both less than 10%. The extraction recovery was between 98.9% and 102.3%. In stability tests (samples stored at room temperature for 4 h, 3 freeze-thaw cycles, -70 ℃ for 7 d and 4 ℃ in autosampler for 24 h), the rates of change of inosine in human plasma were less than 15%.


·The established SPE-HPLC-MS/MS method is simple, rapid, sensitive and accurate, and it is suitable for the clinical monitoring of inosine concentration in human plasma.

Key words: inosine, blood drug level, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), solid-phase extraction (SPE)

CLC Number: