A HPLC–MS method for the simultaneous quantification of fourteen antiretroviral agents in peripheral blood mononuclear cell of HIV infected patients optimized using medium corpuscular volume evaluation

D’Avolio, Antonio; Simiele, Marco; Siccardi, Marco; Baietto, Lorena; Sciandra, Mauro; Oddone, Valentina; Stefani, Francesca Romana; Agati, Silvia; Cusato, Jessica; Bonora, Stefano; Perri, Giovanni Di

doi:10.1016/j.jpba.2010.10.011

Cited by 59 publications

(63 citation statements)

References 64 publications

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“…Although a number of analytical methods have been developed for anti-HIV drugs determination in human plasma (4,10,(16)(17)(18)(19)(20)(21)(22)(23)(24), the HPLC-MS/MS method here reported, using a QqTOF mass analyzer, displays great advantages including the possibility to obtain very high specificity towards the selected analytes (e.g., the anti-HIV drugs considered), despite a simple and rapid sample preparation. This target was achieved by the rational exploitation of the chromatographic and mass-spectrometric tools.…”

Section: Resultsmentioning

confidence: 99%

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

Notari¹,

Sergi²,

Montesano³

et al. 2012

IUBMB Life

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SummaryThe 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 lL. 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-lL 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 3 1.0 mm i.d.) filled with 3-lm C 18 particles. The mobile phase was delivered at 70 lL/ 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. Abbreviations AIDS, acquired immune deficiency syndrome; HAART, highly active anti-retroviral therapy; HIV, human immunodeficiency virus; HPLC-MS/MS, high-performance liquid chromatography-mass spectrometry; LOD, limit of detection; LOQ, limit of quantification; NNRTI, non-nucleoside reverse transcriptase inhibitor; NRTI, nucleoside reverse transcriptase inhibitor; PI, protease inhibitor; QqTOF, quadrupole time-of-flight; SPE, solid-phase extraction; TDM, therapeutic drug monitoring; TOF, time-of-flight.

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Section: Resultsmentioning

confidence: 99%

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

Notari¹,

Sergi²,

Montesano³

et al. 2012

IUBMB Life

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“…The method we've developed and validated, based on a very close PBMC isolation and drug extraction procedure published by our group for antiretroviral drugs [44]. This method can be used everywhere, because require instruments available in all laboratories.…”

Section: Discussionmentioning

confidence: 99%

“…Reliability of our method has been demonstrated for all drug concentrations; relative error at QCs concentrations, intra-day and inter-day precision (Table 3) indicate the good performances of our method. Absence of interference peaks at the analyte retention times, without a "matrix effect", coupled with an experienced collection procedure and treatment of PBMC [44] allowed accurate measurement of drugs intracellular levels.…”

Section: Discussionmentioning

confidence: 99%

“…The six calibration standards and three quality controls (QCs) were prepared adding a determined volume of stock solutions, or diluted stock solution, to each blank PBMCs aliquots before storage at -80°C, during no more than three months, in a manner similar to that described in other publications [43][44].…”

Section: Stock Solutions Standards (Std) and Quality Controls (Qc)mentioning

confidence: 99%

“…PBMCs were isolated from 10 to 14 mL of blood using lymphoprep density gradient centrifugation (700 g, 25 min, 4°C with a Jouan Centrifuge [Model BR4i, Saint-Herblain, France]) at each sampling, as described previously [44]. PBMCs were then fast washed twice in 40 mL cold-ice phosphate-buffered saline and centrifuged (750 g, 6 min, 4°C).…”

Section: Pbmc Isolationmentioning

confidence: 99%

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HPLC–MS method for the simultaneous quantification of the antileukemia drugs imatinib, dasatinib and nilotinib in human peripheral blood mononuclear cell (PBMC)

D’Avolio¹,

Simiele²,

Francia

et al. 2012

Journal of Pharmaceutical and Biomedical Analysis

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A new method using high performance liquid chromatography coupled with electrospray mass spectrometry is described for the quantification of PBMC concentration of tyrosine kinase inhibitors imatinib, dasatinib and nilotinib. A simple PBMC isolation and extraction procedure were applied on 10-14 mL of blood aliquots. Chromatographic separation of drugs and Internal Standard (quinoxaline) was achieved with a gradient (acetonitrile and water + formic acid 0.05%) on a C18 reverse phase analytical column with 25 min of analytical run, at flow rate of 0.25 mL/min. Mean intra-and inter-day precision for all compounds were 8.76 and 12.20%; mean accuracy was -3.86%; extraction recovery ranged within 79 and 91%. Calibration curves ranged from 50.0 to 0.25 ng. The limit of quantification was set at 0.25 ng for all the analyzed drugs.This novel developed methodology allows a specific, sensitive and reliable simultaneous intracellular determination of the three tyrosine kinase inhibitors imatinib, dasatinib and nilotinib in a single chromatographic run, useful for drugs estimation in PBMC of patients affected by chronic myeloid leukemia.

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An Introduction to Tests Performed in Toxicology Laboratories

Dasgupta

2012

Resolving Erroneous Reports in Toxicology and Therapeutic Drug Monitoring

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A HPLC–MS method for the simultaneous quantification of fourteen antiretroviral agents in peripheral blood mononuclear cell of HIV infected patients optimized using medium corpuscular volume evaluation

Cited by 59 publications

References 64 publications

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

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

HPLC–MS method for the simultaneous quantification of the antileukemia drugs imatinib, dasatinib and nilotinib in human peripheral blood mononuclear cell (PBMC)

An Introduction to Tests Performed in Toxicology Laboratories

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