Determination of stavudine in human plasma and urine by high-performance liquid chromatography using a reduced sample volume.

Sensitive high-performance liquid chromatographic assays have been developed for the quantification of stavudine (2',3'-didehydro-3'-deoxythymidine, d4T) in human plasma and urine. The methods are linear over the concentration ranges 0.025-25 and 2-150 microg/ml in plasma and urine, respectively. An aliquot of 200 microl of plasma was extracted with solid-phase extraction using Oasis cartridges, while urine samples were simply diluted 1/100 with HPLC water. The analytical column, mobile phase, instrumentation and chromatographic conditions are the same for both methods. The methods have been validated separately, and stability tests under various conditions have been performed. The detection limit is 12 ng/ml in plasma for a sample size of 200 microl. The bioanalytical assay has been used in a pharmacokinetic study of pregnant women and their newborns.

Specific radioimmunoassays for the measurement of stavudine in human plasma and urine.

Sensitive and specific radioimmunoassays (RIAs) have been developed and validated for the determination of stavudine, a nucleoside analog possessing anti-human immunodeficiency virus (HIV) activity, in human plasma and urine. The hemisuccinate of stavudine was conjugated with histamine and radioiodinated to yield the radiotracer. Antisera were produced by injecting the immunogen, stavudine-hemisuccinate-bovine thyroglobulin, into rabbits. The antisera exhibited high specificity for stavudine as the structurally related analogs and other anti-HIV agents did not interfere in the assays. The methods could reliably quantitate stavudine in plasma from 2.5-100 ng ml-1 and in urine from 5.0-1000 ng ml-1 (after 2.5-fold dilution) with good accuracy and precision. The lower limits of quantitation were 2.5 ng ml-1 in human plasma and 5.0 ng ml-1 in urine (after 2.5-fold dilution). The RIA methods were applied to the analysis of stavudine in plasma and urine obtained from HIV-infected patients receiving the drug in clinical trials.

Prenatal and postpartum pharmacokinetics of stavudine (2',3'-didehydro-3'-deoxythymidine) and didanosine (dideoxyinosine) in pigtailed macaques (Macaca nemestrina).

Stavudine (5 mg/kg of body weight; n = 7) or didanosine (3.2 mg/kg; n = 4) was administered as an intravenous bolus to pregnant pigtailed macaques (Macaca nemestrina) near term and 4 to 5 weeks postpartum. No significant differences were found between the prenatal and postpartum total plasma drug clearance, steady-state volume of distribution, terminal plasma drug half-life, mean body residence time, or recovery of unchanged drug in urine. These data indicate that pregnancy does not affect the pharmacokinetics of stavudine or didanosine in M. nemestrina.

Pharmacokinetics of co-administered didanosine and stavudine in HIV-seropositive male patients.

The pharmacokinetics of single and co-administered didanosine and stavudine were evaluated in 10 HIV-seropositive subjects in an open, within subject design in which each subject received each of three treatments. Single doses of didanosine 100 mg were alternated randomly with single doses of stavudine 40 mg on days 1 and 2. Beginning on day 3, subjects received the same doses of both drugs simultaneously every 12 h for nine doses. Serial blood and urine samples were obtained on single dose days 1 and 2, first simultaneous dose day 3, and last simultaneous dose day 7. The average maximum plasma concentrations of didanosine and stavudine before and after simultaneous administration were 422 +/- 184 (s.d.) ng ml-1 and 603 +/- 160 (s.d.) ng ml-1, and 419 +/- 153 (s.d.) ng ml-1 and 726 +/- 188 (s.d.) ng ml-1, respectively. Didanosine and stavudine AUC values before and after simultaneous administration were 615 +/- 170 (s.d.) ng ml-1 h and 1246 +/- 230 (s.d.) ng ml-1 h, 637 +/- 155 (s.d.) ng ml-1 h and 1326 +/- 267 (s.d.) ng ml-1 h, respectively. No significant changes in maximum plasma concentration, AUC elimination half-life, or renal clearance of didanosine and stavudine were observed when the drugs were administered simultaneously. Co-administration of didanosine 100 mg and stavudine 40 mg is well tolerated and the drugs do not interact pharmacokinetically.