Document Detail

Improved ruggedness of an ion-pairing liquid chromatography/tandem mass spectrometry assay for the quantitative analysis of the triphosphate metabolite of a nucleoside reverse transcriptase inhibitor in peripheral blood mononuclear cells.
MedLine Citation:
PMID:  23280981     Owner:  NLM     Status:  MEDLINE    
RATIONALE: Nucleotide analogs are highly polar and ionic, which impose great challenges on bioanalysis. Ion-pairing liquid chromatography/tandem mass spectrometry (LC/MS/MS) is the predominant reported approach for such compounds. Assay ruggedness of ion-pairing LC/MS/MS methods was often a challenge due to the potential contamination of the ion source of the mass spectrometer and LC column performance deterioration caused by ion-pairing reagents.
METHODS: An ion-pairing reagent was only added to the reconstitution solution to minimize its exposure to the MS ion source. To achieve optimum sensitivity, high pH mobile phases and negative ion ESI were needed for the LC/MS/MS method. However, high pH mobile phases led to the accumulation of ion-pairing reagent on the analytical column, which was washed off with an acidic solution to restore the column performance. In addition, isopropanol was used as a mobile phase modifier to improve peak shape and sensitivity.
RESULTS: The limit of detection was established at 1.0 ng/mL in the cell lysate. The calibration curve showed good linearity over the range of 1.0 to 100 ng/mL. The overall accuracy was no less than 87.7% based on four levels of quality control samples. Inter-run precision and intra-run precision across four analytical runs for low, geometric, medium and high QCs were less than 12.9.
CONCLUSIONS: By identifying and addressing the root cause of the assay ruggedness problem, we have developed a rugged ion-pairing LC/MS/MS method for a triphosphate (TP) metabolite of BMS-986001 in peripheral blood mononuclear cells. The new method overcame challenges such as a rapid deterioration of the peak shape, increased carryover and extremely poor column life. The peak shape was well maintained throughout multiple analytical runs. This method has been successfully applied to a toxicology study in cynomolgus monkey.
Yue Zhao; Guowen Liu; Yifei Liu; Long Yuan; Dara Hawthorne; Jim X Shen; Mausumee Guha; Anne Aubry
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Rapid communications in mass spectrometry : RCM     Volume:  27     ISSN:  1097-0231     ISO Abbreviation:  Rapid Commun. Mass Spectrom.     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-01-02     Completed Date:  2013-06-03     Revised Date:  2013-08-27    
Medline Journal Info:
Nlm Unique ID:  8802365     Medline TA:  Rapid Commun Mass Spectrom     Country:  England    
Other Details:
Languages:  eng     Pagination:  481-8     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 John Wiley & Sons, Ltd.
Bioanalytical Sciences Department, Research and Development, Bristol-Myers Squibb Co.
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MeSH Terms
Chromatography, High Pressure Liquid / methods*,  standards
Drug Stability
Hydrogen-Ion Concentration
Ions / chemistry
Leukocytes, Mononuclear / chemistry*
Macaca fascicularis
Nucleotides / blood*,  chemistry
Organic Chemicals / chemistry
Reproducibility of Results
Reverse Transcriptase Inhibitors / blood*,  chemistry
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization / methods*,  standards
Tandem Mass Spectrometry / methods
Thymidine / analogs & derivatives*,  blood,  chemistry
Reg. No./Substance:
0/BMS-986001; 0/Ions; 0/Nucleotides; 0/Organic Chemicals; 0/Reverse Transcriptase Inhibitors; 50-89-5/Thymidine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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