Fast, simple and cost-effective determination of thiopental in human plasma by a new HPLC technique.

BACKGROUND: Thiopental is an anaesthetic drug that is largely used in both short-term and long-term infusion. After long-term infusion of thiopental, non-linear and inter-individual-dependent pharmacokinetics occur because of the saturation and/or induction of the metabolism. Clinical monitoring is important so that therapeutic adjustments can be made in many of the different pharmacological treatments, especially when long-term infusion is required. We describe a new, rapid HPLC method for the determination of plasma thiopental. METHODS: Sample preparation involved precipitation of plasma proteins using a mixture of methanol, zinc sulfate and ethylene glycol, and containing the internal standard 5-ethyl-5-p-tolyl-barbituric acid. After adding trichloroacetic acid, the sample was centrifuged and the supernatant was injected into a C(18) reversed-phase column. The mobile phase used was water-methanol-acetonitrile (50:40:10, v/v). The eluent was monitored at 290 nm. RESULTS: The calibration curve was linear from 0.2 to 100 microg/mL. Precision, calculated as the coefficient of variation (%), was in the range of 3.62-0.70% for the within-day assay and 5.77-1.51% for the between-day assay. The absolute recoveries obtained from supplemented samples were never less than 100%. CONCLUSIONS: This technique shows good reliability and seems to be suitable for a very fast and simple therapeutic monitoring of plasma thiopental.

Analytical performance of a monoclonal digoxin assay by dry chemistry on the Vitros 950.

Digoxin assay in plasma/serum samples is used for therapeutic measurements as a guide to clinical management of cardiac patients. A thin dry film multilayer monoclonal immunoassay for digoxin, Vitros, was evaluated for analytical performance. The effect of digoxin-like immunoreactive substances (DLIS) was studied assaying plasma samples taken from 100 renal disease patients, 62 hepatic disease patients and 40 pregnant women not receiving digoxin. The Vitros digoxin assay was compared with the AxSym digoxin II assay using plasma samples from 180 patients treated with digoxin. The results revealed satisfactory precision and accuracy for therapeutic drug monitoring purposes: the coefficient of variation (CV%) was lower than 5%; results for dilutions were linear in the range 0.4-3.9 microlg/L and mean analytical recovery was 105%. Measurable DLIS concentrations were observed in 29% of hepatic disease patients and in 7% of renal disease patients with apparent digoxin concentration ranging from 0.4 to 0.75 microg/L. The incidence of DLIS was comparable to that observed with AxSym digoxin II. Comparative results from patient samples gave a regression line equation: Yvitros 950=0.96XAxym +0.14, r=0.89. The data revealed a mean difference of 0.09+/-0.26 microg/L significantly greater than zero (p=0.02). We concluded that Vitros digoxin assay for precision, accuracy and extent of DLIS interference may be a good method for therapeutic drug monitoring; care needs to be taken since assay results generated by Vitros and AxSym analysers are not necessarily interchangeable.

Total carbon dioxide measured by the Vitros enzymatic method.

We evaluated the performance of an enzymatic method using dry chemistry for serum total carbon dioxide (tCO2) determination using a Vitros 500 analyser. Imprecision results were acceptable and the linearity was verified for concentrations within a range of 5.5-39.2 mmol/l, i.e. Y(measured) = 0.93 x(calculated) + 1.32, r = 0.99. The Vitros tCO2 method was unaffected by haemoglobin at all concentrations tested. Significant interference was caused by bilirubin at concentrations higher than 30 mumol/l; the addition of bilirubin lowered the apparent values for tCO2 dose-dependently. Serum tCO2 results were practically the same as those for plasma. The reference interval for venous tCO2 concentrations in a healthy population was: 22.4-34.2 mmol/l (mean: 28.3 mmol/l). Comparison of venous serum tCO2 results assayed using the Vitros method with bicarbonate (HCO3-) values calculated by blood gas determination of pCO2 and pH in arterial blood samples gave poor agreement, r = 0.58. The data revealed a mean difference of 5.48 +/- 3.09 mmol/l between the tCO2 measurements and calculated bicarbonate. This was statistically (p = 0.01) and clinically significant. We conclude that the Vitros method provides reliable tCO2 results in venous serum but this method must not be used as an interchangeable alternative to calculated arterial bicarbonate in order to avoid confusion, misinterpretation of results and erroneous therapeutic decisions.