Simultaneous determination of lamivudine, lopinavir, ritonavir, and zidovudine concentration in plasma of HIV-infected patients by HPLC-MS/MS

Stefania Notari, Manuel Sergi, Camilla Montesano, Jelena Ivanovic, Pasquale Narciso, Leopoldo P. Pucillo, Paolo Ascenzi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The nucleoside reverse transcriptase inhibitors lamivudine and zidovudine and the protease inhibitors lopinavir and ritonavir are currently used in anti-human immunodeficiency virus (HIV) therapy. Here, a high-performance liquid chromatography- mass spectrometry (HPLC-MS/MS) method, using a hybrid quadrupole time-of-flight mass analyzer, is reported for the simultaneous quantification of lamivudine, lopinavir, ritonavir, and zidovudine in plasma of HIV-infected patients. The volume of plasma sample was 600 μL. Plasma samples were extracted by solid-phase using 1 cc Oasis HLB Cartridge (divinylbenzene and N-vinylpyrrolidone) and evaporated in a water bath under nitrogen stream. The extracted samples were reconstituted with 100-μL methanol. Five microliters of the reconstituted samples were injected into a HPLC-MS/MS apparatus, and the analytes were eluted on a Vydac column (250 x 1.0 mm i.d.) filled with 3-μm C 18 particles. The mobile phase was delivered at 70 μL/ min with a linear gradient elution, both acetonitrile and ultrapure water solvents contained 0.2% formic acid. The calibration curves were linear from 0.47 to 20 ng/mL. The absolute recovery ranged between 91 and 107%. The minimal concentration of lamivudine, lopinavir, ritonavir, and zidovudine detectable by HPLC-MS/MS is 0.47, 0.28, 0.30, and 0.66 ng/mL, respectively. The great advantage of the new HPLC-MS/MS method here reported is the possibility to achieve a very high specificity toward the selected anti-HIV drugs, despite the simple and rapid sample preparation. Moreover, this method is easily extendible to the analysis of co-administrated drugs.

Original languageEnglish
Pages (from-to)443-449
Number of pages7
JournalIUBMB Life
Volume64
Issue number5
DOIs
Publication statusPublished - May 2012

Fingerprint

Lopinavir
Ritonavir
Lamivudine
Zidovudine
Viruses
divinyl benzene
formic acid
High Pressure Liquid Chromatography
HIV
Plasmas
Reverse Transcriptase Inhibitors
Water
High performance liquid chromatography
Protease Inhibitors
Nucleosides
Pharmaceutical Preparations
Mass spectrometry
Methanol
Nitrogen
Calibration

Keywords

  • HIV nucleoside reverse transcriptase inhibitors
  • HIV protease inhibitors
  • HPLC-MS/MS
  • Therapeutic drug monitoring

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Clinical Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Simultaneous determination of lamivudine, lopinavir, ritonavir, and zidovudine concentration in plasma of HIV-infected patients by HPLC-MS/MS. / Notari, Stefania; Sergi, Manuel; Montesano, Camilla; Ivanovic, Jelena; Narciso, Pasquale; Pucillo, Leopoldo P.; Ascenzi, Paolo.

In: IUBMB Life, Vol. 64, No. 5, 05.2012, p. 443-449.

Research output: Contribution to journalArticle

Notari, Stefania ; Sergi, Manuel ; Montesano, Camilla ; Ivanovic, Jelena ; Narciso, Pasquale ; Pucillo, Leopoldo P. ; Ascenzi, Paolo. / Simultaneous determination of lamivudine, lopinavir, ritonavir, and zidovudine concentration in plasma of HIV-infected patients by HPLC-MS/MS. In: IUBMB Life. 2012 ; Vol. 64, No. 5. pp. 443-449.
@article{4250a985a4364bb6bb826ca8b695cd73,
title = "Simultaneous determination of lamivudine, lopinavir, ritonavir, and zidovudine concentration in plasma of HIV-infected patients by HPLC-MS/MS",
abstract = "The nucleoside reverse transcriptase inhibitors lamivudine and zidovudine and the protease inhibitors lopinavir and ritonavir are currently used in anti-human immunodeficiency virus (HIV) therapy. Here, a high-performance liquid chromatography- mass spectrometry (HPLC-MS/MS) method, using a hybrid quadrupole time-of-flight mass analyzer, is reported for the simultaneous quantification of lamivudine, lopinavir, ritonavir, and zidovudine in plasma of HIV-infected patients. The volume of plasma sample was 600 μL. Plasma samples were extracted by solid-phase using 1 cc Oasis HLB Cartridge (divinylbenzene and N-vinylpyrrolidone) and evaporated in a water bath under nitrogen stream. The extracted samples were reconstituted with 100-μL methanol. Five microliters of the reconstituted samples were injected into a HPLC-MS/MS apparatus, and the analytes were eluted on a Vydac column (250 x 1.0 mm i.d.) filled with 3-μm C 18 particles. The mobile phase was delivered at 70 μL/ min with a linear gradient elution, both acetonitrile and ultrapure water solvents contained 0.2{\%} formic acid. The calibration curves were linear from 0.47 to 20 ng/mL. The absolute recovery ranged between 91 and 107{\%}. The minimal concentration of lamivudine, lopinavir, ritonavir, and zidovudine detectable by HPLC-MS/MS is 0.47, 0.28, 0.30, and 0.66 ng/mL, respectively. The great advantage of the new HPLC-MS/MS method here reported is the possibility to achieve a very high specificity toward the selected anti-HIV drugs, despite the simple and rapid sample preparation. Moreover, this method is easily extendible to the analysis of co-administrated drugs.",
keywords = "HIV nucleoside reverse transcriptase inhibitors, HIV protease inhibitors, HPLC-MS/MS, Therapeutic drug monitoring",
author = "Stefania Notari and Manuel Sergi and Camilla Montesano and Jelena Ivanovic and Pasquale Narciso and Pucillo, {Leopoldo P.} and Paolo Ascenzi",
year = "2012",
month = "5",
doi = "10.1002/iub.1025",
language = "English",
volume = "64",
pages = "443--449",
journal = "IUBMB Life",
issn = "1521-6543",
publisher = "Wiley-Blackwell",
number = "5",

}

TY - JOUR

T1 - Simultaneous determination of lamivudine, lopinavir, ritonavir, and zidovudine concentration in plasma of HIV-infected patients by HPLC-MS/MS

AU - Notari, Stefania

AU - Sergi, Manuel

AU - Montesano, Camilla

AU - Ivanovic, Jelena

AU - Narciso, Pasquale

AU - Pucillo, Leopoldo P.

AU - Ascenzi, Paolo

PY - 2012/5

Y1 - 2012/5

N2 - The nucleoside reverse transcriptase inhibitors lamivudine and zidovudine and the protease inhibitors lopinavir and ritonavir are currently used in anti-human immunodeficiency virus (HIV) therapy. Here, a high-performance liquid chromatography- mass spectrometry (HPLC-MS/MS) method, using a hybrid quadrupole time-of-flight mass analyzer, is reported for the simultaneous quantification of lamivudine, lopinavir, ritonavir, and zidovudine in plasma of HIV-infected patients. The volume of plasma sample was 600 μL. Plasma samples were extracted by solid-phase using 1 cc Oasis HLB Cartridge (divinylbenzene and N-vinylpyrrolidone) and evaporated in a water bath under nitrogen stream. The extracted samples were reconstituted with 100-μL methanol. Five microliters of the reconstituted samples were injected into a HPLC-MS/MS apparatus, and the analytes were eluted on a Vydac column (250 x 1.0 mm i.d.) filled with 3-μm C 18 particles. The mobile phase was delivered at 70 μL/ min with a linear gradient elution, both acetonitrile and ultrapure water solvents contained 0.2% formic acid. The calibration curves were linear from 0.47 to 20 ng/mL. The absolute recovery ranged between 91 and 107%. The minimal concentration of lamivudine, lopinavir, ritonavir, and zidovudine detectable by HPLC-MS/MS is 0.47, 0.28, 0.30, and 0.66 ng/mL, respectively. The great advantage of the new HPLC-MS/MS method here reported is the possibility to achieve a very high specificity toward the selected anti-HIV drugs, despite the simple and rapid sample preparation. Moreover, this method is easily extendible to the analysis of co-administrated drugs.

AB - The nucleoside reverse transcriptase inhibitors lamivudine and zidovudine and the protease inhibitors lopinavir and ritonavir are currently used in anti-human immunodeficiency virus (HIV) therapy. Here, a high-performance liquid chromatography- mass spectrometry (HPLC-MS/MS) method, using a hybrid quadrupole time-of-flight mass analyzer, is reported for the simultaneous quantification of lamivudine, lopinavir, ritonavir, and zidovudine in plasma of HIV-infected patients. The volume of plasma sample was 600 μL. Plasma samples were extracted by solid-phase using 1 cc Oasis HLB Cartridge (divinylbenzene and N-vinylpyrrolidone) and evaporated in a water bath under nitrogen stream. The extracted samples were reconstituted with 100-μL methanol. Five microliters of the reconstituted samples were injected into a HPLC-MS/MS apparatus, and the analytes were eluted on a Vydac column (250 x 1.0 mm i.d.) filled with 3-μm C 18 particles. The mobile phase was delivered at 70 μL/ min with a linear gradient elution, both acetonitrile and ultrapure water solvents contained 0.2% formic acid. The calibration curves were linear from 0.47 to 20 ng/mL. The absolute recovery ranged between 91 and 107%. The minimal concentration of lamivudine, lopinavir, ritonavir, and zidovudine detectable by HPLC-MS/MS is 0.47, 0.28, 0.30, and 0.66 ng/mL, respectively. The great advantage of the new HPLC-MS/MS method here reported is the possibility to achieve a very high specificity toward the selected anti-HIV drugs, despite the simple and rapid sample preparation. Moreover, this method is easily extendible to the analysis of co-administrated drugs.

KW - HIV nucleoside reverse transcriptase inhibitors

KW - HIV protease inhibitors

KW - HPLC-MS/MS

KW - Therapeutic drug monitoring

UR - http://www.scopus.com/inward/record.url?scp=84859803379&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84859803379&partnerID=8YFLogxK

U2 - 10.1002/iub.1025

DO - 10.1002/iub.1025

M3 - Article

C2 - 22473795

AN - SCOPUS:84859803379

VL - 64

SP - 443

EP - 449

JO - IUBMB Life

JF - IUBMB Life

SN - 1521-6543

IS - 5

ER -