Journal of Shanghai Jiao Tong University (Medical Science) >

2021 , Vol. 41 >Issue 3: 400 - 405

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.03.021

Techniques and methods

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
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  • Department of Pharmacy, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China

Received date: 2020-04-20

  Online published: 2021-04-06

Supported by

Scientific Research Project of Science and Technology Commission of Shanghai Municipality(1741196400)

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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.

Methods

·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.

Results

·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%.

Conclusion

·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)

Cite this article

Guan-qin JIN , Li SUN , Ling-hong XIA , Hou-wen LIN . Determination of inosine in human plasma by high performance liquid chromatography-tandem mass spectrometry combined with solid-phase extraction[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2021 , 41(3) : 400 -405 . DOI: 10.3969/j.issn.1674-8115.2021.03.021

References

1 Ito T, van Kuilenburg AB, Bootsma AH, et al. Rapid screening of high-risk patients for disorders of purine and pyrimidine metabolism using HPLC-electrospray tandem mass spectrometry of liquid urine or urine-soaked filter paper strips[J]. Clin Chem, 2000, 46(4): 445-452.
2 Farthing D, Sica D, Gehr T, et al. An HPLC method for determination of inosine and hypoxanthine in human plasma from healthy volunteers and patients presenting with potential acute cardiac ischemia[J]. J Chromatogr B, 2007, 854(1/2): 158-164.
3 Farthing DE, Farthing CA, Xi L. Inosine and hypoxanthine as novel biomarkers for cardiac ischemia: from bench to point-of-care[J]. Exp Biol Med (Maywood), 2015, 240(6): 821-831.
4 Soares Dos Santos Cardoso F, Cardoso R, Dos Santos Ramalho B, et al. Inosine accelerates the regeneration and anticipates the functional recovery after sciatic nerve crush injury in mice[J]. Neuroscience, 2019, 423: 206-215.
5 黄国华, 薛乾隆, 岳秉宏, 等. 肌苷注射液联合醒脑静治疗DEACMP的临床研究[J]. 西南国防医药, 2019, 29(4): 438-440.
6 唐祯, 徐兰, 陶晓璇, 等. 注射用辅酶A、三磷酸腺苷二钠及肌苷注射液配伍稳定性考察[J]. 中国药师, 2017, 20(1): 184-186.
7 Teixeira FC, Gutierres JM, Soares MSP, et al. Inosine protects against impairment of memory induced by experimental model of Alzheimer disease: a nucleoside with multitarget brain actions[J]. Psychopharmacology (Berl), 2020, 237(3): 811-823.
8 李琴, 毕明俊, 张红, 等. 肌苷对脑缺血再灌注后COX-2表达的调节作用[J]. 青岛大学医学院学报, 2006, 42(1): 13-17.
9 Yuan S, Jiang XF, Zhou XY, et al. Inosine alleviates depression-like behavior and increases the activity of the ERK-CREB signaling in adolescent male rats[J]. Neuroreport, 2018, 29(14): 1223-1229.
10 马全瑞. 肌苷对化学性缺氧损伤的少突胶质细胞保护作用的研究[D]. 西安: 第四军医大学, 2011.
11 Iskierko Z, Sosnowska M, Sharma PS, et al. Extended-gate field-effect transistor (EG-FET) with molecularly imprinted polymer (MIP) film for selective inosine determination[J]. Biosens Bioelectron, 2015, 74: 526-533.
12 Goyal RN, Gupta VK, Chatterjee S. Simultaneous determination of adenosine and inosine using single-wall carbon nanotubes modified pyrolytic graphite electrode[J]. Talanta, 2008, 76(3): 662-668.
13 Farthing D, Xi L, Gehr L, et al. High-performance liquid chromatography (HPLC) determination of inosine, a potential biomarker for initial cardiac ischaemia, using isolated mouse hearts[J]. Biomarkers, 2006, 11(5): 449-459.
14 Stentoft C, Vestergaard M, L?vendahl P, et al. Simultaneous quantification of purine and pyrimidine bases, nucleosides and their degradation products in bovine blood plasma by high performance liquid chromatography tandem mass spectrometry[J]. J Chromatogr A, 2014, 1356: 197-210.
15 Cohen S, Megherbi M, Jordheim LP, et al. Simultaneous analysis of eight nucleoside triphosphates in cell lines by liquid chromatography coupled with tandem mass spectrometry[J]. J Chromatogr B, 2009, 877(30): 3831-3840.
16 Lu JJ, Jia BJ, Yang L, et al. Ultra-high performance liquid chromatography with ultraviolet and tandem mass spectrometry for simultaneous determination of metabolites in purine pathway of rat plasma[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2016, 1036/1037: 84-92.
17 Li L, Xu Y, Wald DN, et al. Determination of 6-benzylthioinosine in mouse and human plasma by liquid chromatography-tandem mass spectrometry[J]. J Chromatogr B, 2009, 877(30): 3871-3877.
18 Marquet P, Saint-Marcoux F, Prémaud A, et al. Performance of the new mycophenolate assay based on IMPDH enzymatic activity for pharmacokinetic investigations and setup of Bayesian estimators in different populations of allograft recipients[J]. Ther Drug Monit, 2009, 31(4): 443-450.
19 Farouk F, ElKady EF, Azzazy HME. Simultaneous UPLC-MS/MS determination of antiepileptic agents for dose adjustment[J]. Biomed Chromatogr, 2017, 31(7). DOI:10.1002/bmc.3921.
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