Biomedical subjects

Faye B Vicente

Publications and source records attributed to Faye B Vicente.

3 recordsLinked to original sources

PubMed2006

Measurement of serum testosterone using high-performance liquid chromatography/tandem mass spectrometry.

Low levels of serum testosterone typically found in women and children cannot be reliably measured by immunoassay. We developed a simple and sensitive method using high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS). Sample preparation involved protein precipitation of serum (1.0 mL) with acetonitrile containing the internal standard (testosterone-d3) followed by liquid extraction with methylene chloride. The chromatographic cycle per specimen was 10 min. The performance was evaluated according to the CLSI EP10-A2 protocol. Within- and between-run imprecision was 20.9%, 2.29% and 1.80%, and 1.81%, 3.58% and 2.97% at mean concentrations of 0.17, 14.1 and 28.8 nM, respectively, with no apparent carryover. The method was linear from 0.21 to 53.1 nM and the analytical recovery was 100.5-106.2% across the concentrations tested. There was no interference observed from other steroids that were tested. Correlation using de-identified patient specimens with a commercial HPLC-MS/MS method showed a slope of 0.991, an intercept of -0.017 and a correlation coefficient (R(2)) of 0.998 by linear regression over concentrations ranging from 0.21 to 16.7 nM. In conclusion, we report here an HPLC-MS/MS method suitable for clinical measurement of serum testosterone.

Chromatography, High Pressure Liquid↗

PubMed2006

Evaluation of an immunoassay of whole blood sirolimus in pediatric transplant patients in comparison with high-performance liquid chromatography/tandem mass spectrometry.

BACKGROUND: Sirolimus is widely used as an immunosuppressant, along with calcineurin inhibitors. Because of its variable pharmacokinetics and narrow therapeutic range, therapeutic drug monitoring of sirolimus is critical to optimize its therapeutic effect and to minimize toxicity. Although liquid chromatography/tandem mass spectrometry is considered the method of choice, the technical and financial challenges of this method are obstacles to its use. A microparticle enzyme immunoassay on the Abbott IMx has recently been reintroduced to the clinical diagnostic market. METHODS: We evaluated this immunoassay using high-performance liquid chromatography/tandem mass spectrometry as the reference method. Precision and carryover were evaluated using an expanded CLSI EP10-A2 protocol. Linearity was studied by serial dilution of high-level whole blood samples, and clinical utility was demonstrated by correlation with the reference method using 56 de-identified pediatric patient samples. RESULTS: The total imprecision was less than 12% across the concentrations tested. The method was linear from 2.6 to 31 nM. The immunoassay showed a mean positive bias of 11.5% in patient specimens relative to high-performance liquid chromatography/mass spectrometry (p<0.001), with a correlation coefficient (R) of 0.953. CONCLUSION: We conclude that the reintroduced immunoassay is useful for therapeutic drug monitoring of sirolimus.

Child↗

PubMed2005

A fast and simple high-performance liquid chromatography/mass spectrometry method for simultaneous measurement of whole blood tacrolimus and sirolimus.

CONTEXT: Combined immunosuppressant therapy using tacrolimus and sirolimus has demonstrable benefits. Simultaneous chromatographic monitoring of whole blood tacrolimus and sirolimus is useful for reducing reagent consumption and turnaround time. We report here a simple and rapid method using high-performance liquid chromatography/mass spectrometry for simultaneous measurement of whole blood tacrolimus and sirolimus. OBJECTIVE: To develop and validate a high-performance liquid chromatography/mass spectrometry method that is suitable for clinical laboratories and that is simple, rapid, and cost-effective. DESIGN: Whole blood (80 microL) was mixed with zinc sulfate solution, followed by protein precipitation with acetonitrile containing the internal standards. After brief centrifugation, the supernatant (20 microL) was injected onto a C18 guard column. The drug and the internal standard ammonium adducts were monitored by multiple reaction monitoring. One-point calibration at levels of 200 ng/mL (249 nM) tacrolimus and 100 ng/mL (109 nM) sirolimus was prepared by adding tacrolimus and sirolimus to immunosuppressant-free whole blood. RESULTS: The assay took 2.5 minutes per sample injection. The total imprecision was between 2.46% and 7.04% for tacrolimus and between 5.22% and 8.30% for sirolimus across the concentrations tested. No carryover was observed, and recoveries were 92% to 98% for tacrolimus and 100% for sirolimus at all levels tested. The tacrolimus was linear from 0.52 to 155.5 ng/mL (0.65-193.4 nM), and sirolimus was linear from 0.47 to 94.8 ng/mL (0.51-103.7 nM). Biases of correlations with commercial methods were within 7%. CONCLUSIONS: This improved method is simple, fast, cost-effective, and suitable for clinical laboratories. It has been implemented for routine clinical monitoring of posttransplantation immunosuppressant therapy.

Chromatography, High Pressure Liquid↗