Rapid and sensitive high-performance liquid chromatographic method for the determination of ritonavir in human plasma.

Ritonavir is an HIV-1 protease inhibitor that is often used to improve the systemic availability of concurrently administered protease inhibitors by impairing their metabolism through cytochrome P450 (CYP) 3A4. Pharmacodynamic relationships between plasma ritonavir concentrations and efficacy and toxicity have also been described. To date, published high-performance liquid chromatographic (HPLC) methods for the determination of ritonavir in human plasma are often complex, requiring the use of a buffered mobile phase that contains amine-modifiers (i.e. diethylamine, triethylamine). In the method herein, ritonavir was precipitated with acetonitrile plus barium hydroxide and zinc sulphate. Chromatographic separation was accomplished using a C-18 base-deactivated (250 x 4.6 mm I.D., 5 atm particle size) analytic column with a mobile phase composed of acetonitrile:water (52:48, v/v). Quantification was performed at 239 nm. Calibration curves were linear from 0.5-25 microg/ml (R2 > 0.999); percent errors, as a measure of accuracy, were < 12.7%. Intra- and inter-assay relative standard deviations (RSD) were below 12.8%. This method provides a rapid and simple means for the accurate and precise analysis of ritonavir in human plasma. Furthermore, the assay requires neither the use of a buffered mobile phase adjusted to a specific pH, nor the addition of amine modifiers. This method has been successfully used to determine plasma ritonavir concentrations in drug interaction studies.

Cardiovascular reactivity to a naturally occurring stressor: development and psychometric evaluation of a psychophysiological assessment procedure.

Three studies were conducted to examine the feasibility, reactive effects of assessment, stability, sampling parameters, and sensitivity of an assessment procedure designed to measure cardiovascular responses to a discrete, naturally occurring, and replicatable stressor--university course examinations. Undergraduate students monitored their blood pressure and heart rate several times during one or two classroom examinations and for several class sessions preceding each examination. Classroom examinations were generally associated with significant increases in subjective measures of distress and cardiovascular measures. Reactive effects of assessment and other sources of error were minimized and responses were reasonably stable over time. These results support the potential utility, validity, and cost-efficiency of this methodology for assessing cardiovascular reactivity to naturally occurring stressors.