Simultaneous determination of 16 anti-HIV drugs in human plasma by high-performance liquid chromatography

Notari, Stefania; Bocedi, Alessio; Ippolito, Giuseppe; Narciso, Pasquale; Pucillo, Leopoldo Paolo; Tossini, Gianna; Donnorso, R. Perrone; Gasparrini, Francesco; Ascenzi, Paolo

doi:10.1016/j.jchromb.2005.12.016

Cited by 126 publications

(97 citation statements)

References 25 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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“…Recently, the combination of EM and TE has demonstrated significantly greater HIV RNA suppression compared to the combination of zidovudine and lamivudine [1][2][3] Several analytical methods that have been reported for the individual determination of TE in biological fluids and pharmaceutical formulations which include liquid chromatography coupled with spectrofluorimetric, UV, and mass spectroscopy detection [4][5][6][7][8][9][10] . For EM several analytical methods have been reported for its individual analysis which includes chiral liquid chromatography, liquid chromatography with UV detection [11][12][13] . Few Bioanalytical methods are reported for combination of TE and EM which includes liquid chromatography with PDA and UV detection 14,15 .…”

Section: Tenofovir (Te; 9-[(r)-2-[[bis[[(isopropoxycarbonyl) Oxy] Metmentioning

confidence: 99%

Simultaneous Determination of Emtricitabine and Tenofovir by Area Under Curve and Dual Wavelength Spectrophotometric Method

Ghorpade¹,

Sali²,

Kategaonkar³

et al. 2010

J. Chil. Chem. Soc.

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Two methods for the simultaneous determination of Emtricitabine and Tenofovir by spectroscopy have been developed. These two simple, accurate and precise methods include Area Under the Curve (AUC) method and Dual Wavelength Method. From a solvent effect studies and the spectral behaviours of Emtricitabine and Tenofovir, methanol was selected as solvent. Emtricitabine shows maximum absorbance at 281 nm and Tenofovir shows maximum absorbance at 259 nm. For the AUC method, the wavelength ranges between 242-248 nm and 269-275 nm were selected with reference to the absorbance curves plotted between the wavelengths of 200-400 nm. In the second method i.e. Dual method in which two wavelengths were selected for each drug in a way so that the difference in absorbance is zero for another drug. Emtricitabine shows equal absorbance at 230.696 nm and 250 nm, where the differences in absorbance were measured for the determination of Tenofovir. Similarly, differences in absorbances at 250 nm and 268.670 nm were measured for determination of Emtricitabine. These methods allows rapid analysis of two drug combination. The results of analysis were validated statistically and by recovery studies. This tablet containing both drugs was assayed using the methods developed, showing a good accuracy and precision.

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“…On the contrary, reverse phase highperformance liquid chromatographic with ultraviolet detector (RP-HPLC-UV) method is feasible due to its easy accessibility and low cost. Also, many HPLC-UV methods for determination of plasma efavirenz (23,(26)(27)(28)(29)(30)(31) or simultaneous determination with other drugs such as anti-tuberculosis (anti-TB) agents or other antiretroviral agents (24,(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47)(48)(49)(50) have been reported. In most of these methods, the volumes of plasma used range from 200 to 900 μL (23,(26)(27)(28)33,34,(44)(45)(46), which were inapplicable for children due to the scarcity of sample.…”

Section: Introductionmentioning

confidence: 99%

A simple, rapid, economical, and practical method for the determination of efavirenz in plasma of Chinese AIDS patients by reverse phase high-performance liquid chromatography with ultraviolet detector

Yin

Meng

Dong

et al. 2014

BST

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Simultaneous determination of 16 anti-HIV drugs in human plasma by high-performance liquid chromatography

Cited by 126 publications

References 25 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

Simultaneous Determination of Emtricitabine and Tenofovir by Area Under Curve and Dual Wavelength Spectrophotometric Method

A simple, rapid, economical, and practical method for the determination of efavirenz in plasma of Chinese AIDS patients by reverse phase high-performance liquid chromatography with ultraviolet detector

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