Simple and rapid determination of nevirapine in human serum by reversed-phase high-performance liquid chromatography

López, Rosa María; Pou, Leonor; Gómez, M. Ramírez; Ruíz, Isabel; Monterde, J.

doi:10.1016/s0378-4347(00)00476-x

Cited by 33 publications

(11 citation statements)

References 5 publications

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“…Two methods of DNA isolation were used in this study. One was the QIAamp DNA Blood Midi Kit (Qiagen, Inc.), which has been widely used for isolating circulating DNA from serum or plasma (1)(2)(3)(4)(5). The other is the modified Guanidine/Promega Resin (G/R) method that was developed to isolate DNA from urine (6 ).…”

Section: Clinical Chemistry 50 No 1 2004mentioning

confidence: 99%

Effects of Concentration and Temperature on the Stability of Nevirapine in Whole Blood and Serum

Bennetto¹,

King²,

Turner³

et al. 2004

Clinical Chemistry

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In sub-Saharan Africa, India, Southeast Asia, and other resource-poor regions of the world, the antiretroviral nonnucleoside reverse transcriptase inhibitor nevirapine (NVP) is widely prescribed for HIV treatment and prevention because of its relative safety, long half-life, and low cost. Furthermore, pregnant women are often prescribed NVP therapy to decrease the rate of mother-tochild HIV transmission or as part of a combination therapeutic regimen in more developed countries. As one example, measuring NVP concentrations in pregnant women and their infants, in particular, has increased as clinics in developing countries attempt to better assess NVP "coverage" (e.g., universal treatment regardless of HIV status vs targeted treatment) (1 ). Because most developing countries currently do not have state-of-theart sample collection, processing, and/or storage capabilities, more data are needed on how various procedures in these countries may affect NVP stability.In clinical chemistry, drug stability during sample processing and storage is an important consideration for the interpretation of drug concentrations. If drugs do not remain stable in a given matrix under the conditions used for storage and/or processing, inaccurate drug concentrations will be obtained. Blood samples taken in resourcepoor regions are often sent to laboratories in the developed world for drug measurements. However, these samples may not be processed and handled in the same manner as they would be in the US. In resource-poor regions, blood is often collected in red-top (no additive) tubes to separate serum from cells. Samples may not be placed on ice immediately but instead left on the laboratory bench for a period of up to 24 h before packaging and shipping for antiretroviral analysis. Furthermore, because of clinical settings in tropical climates, samples may be exposed to high temperatures for an undisclosed amount of time. Currently, NVP stability data under these conditions are not available. The objective of the present study was to assess the short-term stability of NVP in human whole blood and serum as a function of storage time, concentration, and temperature.Four drug-free samples of EDTA-whole blood, heparinwhole blood, and anticoagulant-free serum (25 mL) were obtained, with each sample coming from a different patient. NVP was then added to all three sample types at low (250 g/L) and high (2500 g/L) concentrations. The NVP-enriched samples were subsequently divided and transferred to three environments: the laboratory bench at room temperature (20 -25°C), the refrigerator (4°C), and an incubator (37°C) for a 24-h period. At each time point, (0, 1, 2, 4, 8, or 24 h), 1 mL of NVP-enriched blood product (EDTA and heparin) was removed from the samples stored at each temperature and centrifuged (15 000g for 5 min) to separate the plasma. Three 100-L aliquots of the separated plasma from whole blood or anticoagulant-free serum were taken, and 50-L of internal standard was added to each aliquot. Internal standards were 20 mg/L and 2 mg/...

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Section: Clinical Chemistry 50 No 1 2004mentioning

confidence: 99%

Effects of Concentration and Temperature on the Stability of Nevirapine in Whole Blood and Serum

Bennetto¹,

King²,

Turner³

et al. 2004

Clinical Chemistry

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“…The ultraviolet (UV) wavelength at 284 nm was chosen because there was less interference by endogenous substances in plasma, in spite of the highest W-1 chromatographic peak being observed at 221 nm. Moreover, several compounds were tested for internal standard (IS), including amiodarone hydrochloride (similar to W-1 which was a containing iodic atomic compound; Karmarkar et al, 2014), nevirapine (another non-nucleoside reverse transcriptase inhibitor, NNRTI; Lopez et al, 2001;Rezk et al, 2002) and megestrol acetate (the UV absoption at 280 nm; Farinha et al, 2000;Mortensen and Pedersen, 2007;Seo et al, 2013). Megestrol acetate was chosen as an IS for its appropriate UV-absorption and retention time and high extraction recovery.…”

Section: Methods Development and Optimizationmentioning

confidence: 99%

Development and validation of a high performance liquid chromatography method for determination of 6‐benzyl‐1‐benzyloxymethyl‐5‐iodouracil (W‐1), a novel non‐nucleoside reverse transcriptase inhibitor and its application to a pharmacokinetic study in rats

Wang

et al. 2015

Biomedical Chromatography

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A sensitive and selective high-performance liquid chromatographic (HPLC) method for determination of 6-benzyl-1-benzyloxymethyl-5-iodouracil (W-1), a novel non-nucleoside reverse transcriptase inhibitor in rat plasma, was developed and validated. Chromatographic separation of W-1 and megestrol acetate (internal standard) was achieved on a reversed-phase C18 column at 25°C. The mobile phase was consisted of acetonitrile-water (60:40, v/v) and pumped at a flow rate of 1.0 mL/min. The ultraviolet (UV) detector was set at the absorption wavelength of 284 nm. The calibration curve for W-1 was linear over the concentration range of 0.01-8 µg/mL and the lower limit of quantification was 10 ng/mL. The intra- and inter-day precision and accuracy were <8.9 and 5.3%, respectively. The extraction recoveries ranged from 97.9 to 101.6%. The validated HPLC method was successfully applied to a pharmacokinetic study of W-1 in rats.

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“…Therapeutic drug monitoring and novel extensive drug combinations in acquired immune deficiency syndrome therapy over the past 10 years has produced numerous serum and plasma nevirapine assays using liquid chromatography-ultraviolet detection [10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25], gas chromatography [26], electrokinetic chromatography [27], thin-layer chromatography [28,29], liquid chromatography with tandem mass spectrometry assays [30,31], and an enzyme immunoassay [32]. This paper describes for the first time a sensitive, specific, and rapid LC/MS/MS method for the simultaneous determination of the five nevirapine metabolites in human plasma.…”

Section: Introductionmentioning

confidence: 99%

Quantitation of five nevirapine oxidative metabolites in human plasma using liquid chromatography–tandem mass spectrometry

Rowland

MacGregor

Campbell

et al. 2007

Journal of Chromatography B

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Simple and rapid determination of nevirapine in human serum by reversed-phase high-performance liquid chromatography

Cited by 33 publications

References 5 publications

Effects of Concentration and Temperature on the Stability of Nevirapine in Whole Blood and Serum

Effects of Concentration and Temperature on the Stability of Nevirapine in Whole Blood and Serum

Development and validation of a high performance liquid chromatography method for determination of 6‐benzyl‐1‐benzyloxymethyl‐5‐iodouracil (W‐1), a novel non‐nucleoside reverse transcriptase inhibitor and its application to a pharmacokinetic study in rats

Quantitation of five nevirapine oxidative metabolites in human plasma using liquid chromatography–tandem mass spectrometry

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