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Biomedical subjects

J R Charlson

Publications and source records attributed to J R Charlson.

4 recordsLinked to original sources

A solid phase extraction method for determination of nicotine in serum and urine by isotope dilution gas chromatography/mass spectrometry with selected ion monitoring.

A rapid method for measuring nicotine concentration in serum and urine is described. Deuterated nicotine is used as an internal standard. Nicotine and deuterated nicotine are extracted using a copolymeric-bonded phase silica column. The extract is analysed by gas chromatography coupled with mass spectrometry (GC/MS) operating in selected ion monitoring mode. The method has a lower limit of detection of approximately 2 micrograms/L and is linear to at least 2000 micrograms/L. Within-run percentage coefficients of variation (% CV) are < 4 in both assays over a nicotine concentration range of 10-2000 micrograms/L. Between-run % CV in the serum assay are 5.4, 5.2, 4.8 and 5.9, respectively, at nicotine concentrations of 10, 15, 25, and 50 micrograms/L. Between-run % CV in the urine assay are 5.9, 4.5, 2.7 and 5.2, respectively, at nicotine concentrations of 100, 250, 500, and 2000 micrograms/L. The absolute recovery of nicotine is 61 +/- 6% (mean +/- SD) over the range of 10-250 micrograms/L. The assay has been used to measure serum nicotine concentrations and 24-h urinary excretion of nicotine to monitor the extent of replacement in subjects receiving transdermal nicotine therapy for smoking cessation.

Deuterium↗

Automated HPLC assay of fluoxetine and norfluoxetine in serum.

This automated assay determines the concentration of the antidepressant fluoxetine (Prozac) and its active metabolite norfluoxetine in serum by reversed-phase HPLC with spectrophotometric detection. Extraction, injection, and quantification are performed by the Gilson Aspec automated sample handler. The patient's specimen, with added protriptyline as internal standard, is extracted with solid-phase and liquid-liquid methods. Separation is conducted isocratically on a 5-microns (particle size) Supelcosil LC-8-DB reversed-phase column with a triethylamine acetate:acetonitrile mobile phase. The detection limit is 10 micrograms/L and absorbance varies linearly with concentration between 20 and 1,000 micrograms/L for both compounds. Mean recovery was 62% for fluoxetine and 70% for norfluoxetine over linear limits. Within-run and day-to-day imprecision (CV), evaluated at three concentrations (50, 200, and 500 micrograms/L), ranged from 2% to 7%. An extensive interference study of 108 drugs was conducted. Results (n = 58) by the automated method (y) correlated well with those by a manual HPLC method (x): y = 0.96x + 10.20 (r = 0.951, Sylx = 42.9) for fluoxetine, and y = 0.95x - 1.37 (r = 0.917, Sylx = 47.2) for norfluoxetine.

Autoanalysis↗

Marijuana testing--how good is it?

Testing for the use of marijuana necessitates one or more levels of assessment, depending on the clinical application of the test results. The EMIT-dau screening test with a sensitivity limit of 20 ng/ml is highly satisfactory for screening for the absence of marijuana in the urine. This test has a virtual 100% true-negative rate as long as an unadulterated urine specimen is analyzed. Positive results by the EMIT-dau procedure are presumptive because the test seems to produce false-positive results when applied to a random population of suspected drug users. In a population in a specific clinical environment such as in drug-treatment programs in which 20 of every 100 specimens will yield positive THC-COOH results by EMIT-dau screening, 3 (15%) of the 20 positive results will likely be false-positive. Our experience with the EMIT-dau suggests that of 100 test results, 3 will be false-positive, an overall 3% false-positive rate. If a positive test result will put the patient in considerable jeopardy and the screening result is the only evidence of drug use, confirmatory testing is imperative. Of the confirmatory tests, GC/MS seems to have the specificity necessary to provide a high level of confidence in the results. A combination of the EMIT procedure with a sensitivity level of 20 ng/ml and GC/MS confirmation yields virtually 100% accuracy in detection of marijuana abuse.

Cannabinoids↗

Improved chromatography of cyclosporine.

A fast liquid chromatographic procedure for the measurement of whole-blood cyclosporine concentration is described. A sample preparation using solid-phase extraction requires 5.5 min of technical effort per sample. Chromatographic development is complete in 6 min. The results of this procedure correlate with those of a previously validated procedure. The slope of the correlation equation is 1.04, the gamma-intercept is -17, and the correlation coefficient is 0.99. Reproducibility of the procedure at midtherapeutic concentration is 5.5%, and sensitivity is 25 ng/ml. No chemical interference was observed from 33 drugs commonly co-administered during immunosuppression.

Chromatography, Liquid↗