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

Abstract

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.

Hollow mesoporous structured molecularly imprinted polymer as adsorbent in pipette‐tip solid‐phase extraction for the determination of antiretrovirals from plasma of HIV‐infected patients

“…The literature is lacking in studies involving the use of MIP for extraction of ARVs from human plasma. A typical comparison with other methods reported in the literature using SPE for the determination of 3TC, AZT and EFZ in different matrices is presented in Supporting Information Table 3 , which confirm that LOQ value is appropriate and comparable with other research works. The low LOQs are mainly attributed to the use of highly sensitive, complex and expensive detector, i.e.…”

Creation of a Long-Acting Nanoformulated 2′,3′-Dideoxy-3′-Thiacytidine

“…A hydrophobically modified 3TC derivative (designated M3TC) was synthesized according to the scheme illustrated in Figure 1 and described in the Supplemental Digital Content, 31 http://links.lww.com/QAI/A900 . M3TC nanoparticles were formulated by high-pressure homogenization (Avestin EmulsiFlex-C3; Avestin Inc., Ottawa, ON, Canada), using P407 and folate-decorated P407 (FA-P407) to encase the drug crystals.…”

Bio-analytical Assay Methods used in Therapeutic Drug Monitoring of Antiretroviral Drugs-A Review

“…This RP-HPLC assay allows simultaneous quantification of six PIs [IDV, RTV, SQV, NFV, APV, and LPV] using carbamazepine as an internal standard. Thereafter, several simultaneous assays on HPLC with UV PDA, Diode array and MS detectors have been made available in the literature [197][198][199][200][201][202][203][204][205][206][207][208][209][210][211][212][213][214][215][216]. Although not much in use, Azoulay et al in 2004 reported an immunoassay for LPV [217].…”