A novel ultrasensitive LC-MS/MS assay for quantification of intracellular raltegravir in human cell extracts

Brian L. Robbins; Sarah R. Nelson; Courtney V. Fletcher

doi:10.1016/j.jpba.2012.05.035

A novel ultrasensitive LC-MS/MS assay for quantification of intracellular raltegravir in human cell extracts

Brian L. Robbins, Sarah R. Nelson, Courtney V. Fletcher

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

An assay using ultrahigh pressure liquid chromatography and mass spectrometry detection was developed and validated for measurement of the HIV integrase inhibitor raltegravir (MK-0518) in human cell extracts. The assay is designed to utilize 200μl of 70% MeOH cell extract derived from human peripheral blood mononuclear cells or human tissue samples. The assay is linear over a range from 0.0023 to 9.2ngml^-1. The average %CV (SD/Mean)100 and %deviation ((observed-target)/target)100 were less than 20% at the lower limit of quantification and less than 15% over the range of the curve. This assay is an accurate and highly sensitive method for determining raltegravir concentrations in cellular extracts with a lower limit 40 to over 100-fold lower than other methods in the literature. We also present a new processing method where a rapid spin through oil produced a significant increase in apparent intracellular raltegravir concentration compared with conventional processing.

Original language	English (US)
Pages (from-to)	378-387
Number of pages	10
Journal	Journal of Pharmaceutical and Biomedical Analysis
Volume	70
DOIs	https://doi.org/10.1016/j.jpba.2012.05.035
State	Published - Nov 2012

Keywords

HIV
Integrase inhibitor
Intracellular
LC-MS/MS
Raltegravir

ASJC Scopus subject areas

Analytical Chemistry
Pharmaceutical Science
Drug Discovery
Spectroscopy
Clinical Biochemistry

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10.1016/j.jpba.2012.05.035

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title = "A novel ultrasensitive LC-MS/MS assay for quantification of intracellular raltegravir in human cell extracts",

abstract = "An assay using ultrahigh pressure liquid chromatography and mass spectrometry detection was developed and validated for measurement of the HIV integrase inhibitor raltegravir (MK-0518) in human cell extracts. The assay is designed to utilize 200μl of 70% MeOH cell extract derived from human peripheral blood mononuclear cells or human tissue samples. The assay is linear over a range from 0.0023 to 9.2ngml-1. The average %CV (SD/Mean)100 and %deviation ((observed-target)/target)100 were less than 20% at the lower limit of quantification and less than 15% over the range of the curve. This assay is an accurate and highly sensitive method for determining raltegravir concentrations in cellular extracts with a lower limit 40 to over 100-fold lower than other methods in the literature. We also present a new processing method where a rapid spin through oil produced a significant increase in apparent intracellular raltegravir concentration compared with conventional processing.",

keywords = "HIV, Integrase inhibitor, Intracellular, LC-MS/MS, Raltegravir",

author = "Robbins, {Brian L.} and Nelson, {Sarah R.} and Fletcher, {Courtney V.}",

note = "Funding Information: Grant support: supported in part by: P01 AI074340 and RO1AI093319 (to CVF) from the National Institute of Allergy and Infectious Diseases . ",

year = "2012",

month = nov,

doi = "10.1016/j.jpba.2012.05.035",

language = "English (US)",

volume = "70",

pages = "378--387",

journal = "Journal of Pharmaceutical and Biomedical Analysis",

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AU - Robbins, Brian L.

AU - Nelson, Sarah R.

AU - Fletcher, Courtney V.

N1 - Funding Information: Grant support: supported in part by: P01 AI074340 and RO1AI093319 (to CVF) from the National Institute of Allergy and Infectious Diseases .

PY - 2012/11

Y1 - 2012/11

N2 - An assay using ultrahigh pressure liquid chromatography and mass spectrometry detection was developed and validated for measurement of the HIV integrase inhibitor raltegravir (MK-0518) in human cell extracts. The assay is designed to utilize 200μl of 70% MeOH cell extract derived from human peripheral blood mononuclear cells or human tissue samples. The assay is linear over a range from 0.0023 to 9.2ngml-1. The average %CV (SD/Mean)100 and %deviation ((observed-target)/target)100 were less than 20% at the lower limit of quantification and less than 15% over the range of the curve. This assay is an accurate and highly sensitive method for determining raltegravir concentrations in cellular extracts with a lower limit 40 to over 100-fold lower than other methods in the literature. We also present a new processing method where a rapid spin through oil produced a significant increase in apparent intracellular raltegravir concentration compared with conventional processing.

AB - An assay using ultrahigh pressure liquid chromatography and mass spectrometry detection was developed and validated for measurement of the HIV integrase inhibitor raltegravir (MK-0518) in human cell extracts. The assay is designed to utilize 200μl of 70% MeOH cell extract derived from human peripheral blood mononuclear cells or human tissue samples. The assay is linear over a range from 0.0023 to 9.2ngml-1. The average %CV (SD/Mean)100 and %deviation ((observed-target)/target)100 were less than 20% at the lower limit of quantification and less than 15% over the range of the curve. This assay is an accurate and highly sensitive method for determining raltegravir concentrations in cellular extracts with a lower limit 40 to over 100-fold lower than other methods in the literature. We also present a new processing method where a rapid spin through oil produced a significant increase in apparent intracellular raltegravir concentration compared with conventional processing.

KW - HIV

KW - Integrase inhibitor

KW - Intracellular

KW - LC-MS/MS

KW - Raltegravir

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ER -

A novel ultrasensitive LC-MS/MS assay for quantification of intracellular raltegravir in human cell extracts

Abstract

Keywords

ASJC Scopus subject areas

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