[ Pharmaceutical Sciences Asia - ONLINE ]
Print ISSN 2586-8195 E-ISSN 2586-8470
[ Journal Abbreviation: Pharm.Sci.Asia ]
Mahidol University Journal of Pharmaceutical Sciences
  FORMER NAME   "Mahidol University Journal of Pharmaceutical Sciences" Published Since 1974

 
Abstracts

DOI: https://doi.org/10.14456/mujps.2015.20Pharm Sci Asia 2015; 42(4), 162-168
 

Determination of inosine 5’-monophosphate dehydrogenase activity by high performance liquid chromatography in comparison with normalization methods

MT. Nguyen1,2,3, NT. Tran1, M. Vincent2,3, A. Capron2,3, M. Mourad4, P. Wallemacq2,3*

1 Biochemistry Department, University of Medicine and Pharmacy, Ho Chi Minh city, Viet Nam
2 Clinical Chemistry Department, Cliniques universitaires St Luc, Brussels, Belgium
3 Louvain center for Toxicology and Applied Pharmacology, Universit? catholique de Louvain, UCL, Brussels, Belgium
4 Surgery and abdominal transplantation department, Cliniques universitaires St Luc, Brussels, Belgium


Mycophenolic acid (MPA) is a potent, selective and uncompetitive inhibitor of the inosine 5’-monophophate dehydrogenase (IMPDH), which is the rate-limiting enzyme in the de novo synthesis of guanosine nucleotides being required during T and B lymphocyte proliferation. In this study, HPLC method was developed and validated to determine the IMPDH activity in human peripheral blood mononuclear cells (PBMCs) for the purpose of investigating the effect of MPAon lymphocytes. PBMCs were isolated from total blood samples. IMPDH activity in PBMCs was expressed according to two normalisation methods, namely as the ratio of produced xanthosine monophosphate (XMP) (micromoles) versus either adenosine monophosphate (AMP) (moles), or protein (mg) concentration per incubation time. IMPDH activities were assessed in 41 healthy volunteers. The total HPLC run time was 14 min. This method was linear in the range of 0.5-80 ?mol/L for both AMP and XMP. Mean intra-assay and inter-assay precision and accuracy at quality control (QC) levels were > 94% for AMP and XMP. The recovery was > 75% for AMP and XMP. The lower limit of quantification of both AMP and XMP was 0.5 ?mol/L. No interference was identified in the assay. Both IMPDH activity normalisation methods in our study displayed good and similar results, however, the protein normalisation method displayed slightly better reproducibility in our population cohort.


Keyword:

inosine 5’-monophosphate dehydrogenase activity, mycophenolic acid, HPLC, peripheral blood mononuclear cells




Download full paper (PDF File size: 701.19 KB.)





Vol.42
No.4
October - December 2015

See other volume

 


Vol.42
No.3
July - September 2015

See other volume

 


Vol.42
No.2
April - June 2015

See other volume

 


Vol.42
No.1
January - March 2015

See other volume

 
 
 

Home
Aims and Scope
Editorial Board
Publication Ethics
Instruction to Authors
Announcement
All Volumes & Issues
Submit Online
Contact us
   
Search
   
Faculty of Pharmacy Mahidol University
Mahidol University
Member's menu

You're now loging on with following account:
Guest (unknown)

Please Sign in or Register



    Copyright © 2017
    Faculty of Pharmacy, Mahidol University, THAILAND