The routine analysis of breast milk for drugs of abuse in a clinical toxicology laboratory.

Drug screening of breast milk in a clinical toxicology laboratory is reported. Findings from three cases include cocaine, ethylbenzoylecgonine (cocaethylene), ethanol, oxycodone, codeine, and nicotine. We believe this to be the first report of ethylbenzoylecgonine in human breast milk. One other specimen submitted for analysis was screened with negative results. Screening and confirmation procedures adapted for use with breast milk are described. Finally, the potential for cocaine intoxication from mother to baby is discussed. Estimates of infant blood cocaine concentration are given which may increase awareness of the need to monitor milk and blood cocaine concentrations in the infant when the situation warrants.

Impact of food on the bioavailability of encainide.

The bioavailability of drugs that undergo extensive presystemic hepatic metabolism may be increased by concomitant ingestion with food. The effect of food on the bioavailability of encainide, a class IC antiarrhythmic agent, was evaluated in 14 healthy subjects in this randomized crossover study. The subjects received encainide 35 mg every 8 hours for 7 days and were randomized to receive their test dose of encainide with food or after an overnight fast. Encainide area-under-the-concentration versus time curve (AUCs) were detectable in 3 of 14 subjects after fasting and in 7 of 14 after feeding. Although food increased the mean encainide AUC by more than threefold, this increase did not reach statistical significance because of the large number of subjects with indeterminate encainide AUCs. Food did significantly increase the AUC of O-demethyl-encainide (ODE), but not the AUC of methoxy-O-demethyl-encainide (MODE). Despite the increase in ODE AUC, no significant effect on the surface electrocardiogram 2 hours after dose administration could be detected. Food may increase the bioavailability of encainide and one of its active metabolites (ODE). The clinical relevance of this pharmacodynamic effect warrants further evaluation.

Urinalysis of alpha-hydroxyalprazolam, alpha-hydroxytriazolam, and other benzodiazepine compounds by GC/EIMS.

This procedure was developed as an overall benzodiazepine confirmation scheme and includes the detection of the most important urinary analytes encountered by clinical toxicology laboratories in North America: alpha-hydroxyalprazolam, alpha-hydroxytriazolam, 2-hydroxyethylflurazepam, oxazepam, temazepam, and lorazepam. Desmethyldiazepam (nordiazepam) was not targeted because it is metabolized to oxazepam. This procedure takes advantage of beta-glucuronidase hydrolysis for analysis of intact benzodiazepine molecules, oxazepam-2H5 as an internal standard, a newly developed extraction solvent, and a silylating moiety that may be more sensitive than trimethylsilyl (-TMS) derivatives, the tertbutyldimethylsilyl (-TBDMS) derivative. For all compounds the extraction efficiency was greater than 90% and the limit of quantitation (LOQ at a S/N of 10) was less than 10 ng/mL. Coefficients of variation for a 200-ng/mL control were less than 5% and less than or equal to 11% for within-run and between-run trials, respectively. Of 13 human specimens screened by EMIT and most with self-reported histories, alpha-hydroxyalprazolam was found in seven (range 49-1264 ng/mL), oxazepam was found in five (72-3897 ng/mL), and lorazepam (476 ng/mL), 2-hydroxyethylflurazepam (2301 ng/mL), and alpha-hydroxytriazolam (106 ng/mL) in one each.

Computer-assisted work station timing analysis of instrument labor efficiency.

Labor use ratings assigned to instruments by the Workload Recording Method (WRM) do not change with batch size or walk-away time use. The authors evaluated the effect of both on the labor use of the analyzers Paramax B6100 (Baxter Paramax, Irvine, CA) and Ektachem 700 (Eastman Kodak, Rochester, NY) by timing all worked and walk-away intervals on both instruments. Extrapolation of the data to a workload of slightly more than 1.1 million tests showed that reapportionment of tests to various batch sizes caused Paramax-Ektachem labor cost differences to fluctuate between $37,254 and $34,995. When the minimum usable walk-away interval length was varied from 1 to 20 minutes, Ektachem savings over Paramax increased from $8,700 to $61,400. The WRM predicted a constant $29,050 labor cost advantage for Ektachem over Paramax. If other instruments show similar labor use characteristics with respect to batch size and walk-away utility, laboratory managers who do not consider these factors may fail to select the most cost-effective instruments for their laboratories.

Influence of mexiletine on the pharmacokinetics of theophylline in healthy volunteers.

Preliminary reports suggest an interaction exists between theophylline and mexiletine. We conducted a two-way crossover study in 15 healthy male subjects to assess the magnitude of the pharmacokinetic interaction between mexiletine and theophylline. Twelve subjects completed 5 days of therapy on sustained-release theophylline 200 mg every 12 hours alone and 5 days of therapy with theophylline and mexiletine 150 mg every 8 hours. The two treatment periods were separated by a minimum of 7 days. On the morning of day 5 of each treatment period, blood samples for theophylline to be assayed by fluorescence immunoassay were collected over 24 hours. Mexiletine significantly increased the mean AUC, Cmax, t1/2 beta, and Cl of theophylline. Mexiletine did not affect tmax or Vd. Side effects occurred in 4 subjects during treatment with theophylline alone all of which were judged to be mild in intensity. During concomitant theophylline-mexiletine therapy, 10 subjects reported side effects of which 4 were judged to be severe, 1 moderate, and 5 mild. The magnitude of the increase in theophylline plasma concentrations induced by mexiletine as measured by AUC (0-24) was 58%, which is similar to other preliminary reports of this interaction. The authors conclude that the magnitude of the pharmacokinetic interaction between theophylline and mexiletine may be clinically significant in patients in light of the increased incidence of side effects in our healthy subjects. Theophylline dosage adjustments will be required in patients who receive concomitant mexiletine therapy.

Calibration as a source of imprecision in cholesterol testing. Ramifications for patient risk classification.

Routine calibration of a cholesterol assay system may compromise rather than improve precision. We compared an enzymatic assay on a centrifugal analyzer using a fixed factor with a factor recalculated from the response of standards assayed with each run. Over 36 batch runs, using three quality control materials, we found no statistically significant difference between the two methods in mean value, but in every case the fixed factor values were significantly more precise. With the risk classification system in effect at the time of the study, 32 (9.4%) of 342 patient serum specimens assayed for cholesterol were classified differently based solely on the method of data reduction. Thus, recalibration of our cholesterol assay system contributed to greater imprecision and to discrepancies in classification of patients' risk levels.

Clinical and analytical evaluation of kits for measurement of creatine kinase isoenzyme MB.

We studied the analytical and clinical performance of six methods for creatine kinase (EC 2.7.3.2) isoenzyme MB (CK-MB): three immunoassays (Behring, Hybritech, and International Immunoassay Labs); one immunoinhibition assay (Roche); one immunoinhibition/column method (Du Pont); and one electrophoretic method (Beckman). Between-day precision for all kits was poor at the upper reference limit. All methods gave results linearly related to CK-MB concentration and all were free from CK-MM, CK-BB, and adenylate kinase interference. Only the Du Pont method was adversely affected by atypical isoenzymes. For diagnosis of acute myocardial infarction in a coronary care population (n = 40; prevalence = 45%), all methods were approximately 95% efficient, when appropriate reference criteria were used. Some manufacturers fail to provide data for an appropriate (acutely ill, non-infarct) reference population; decreased diagnostic specificity may result from use of reference ranges based on results for healthy subjects. Expression of CK-MB as a percent of total CK degrades efficiency unless total CK is markedly increased.

Measurement of amikacin in serum by a latex agglutination inhibition test.

Levels of amikacin in serum were determined in 106 serum specimens by a latex agglutination inhibition card test and by radioimmunoassay (RIA). Linear regression analysis demonstrated a high degree of correlation between the two assays (latex = 0.95 (RIA) + 0.69; r = 0.97). Assay of three control sera containing 7.5, 15, and 30 micrograms of amikacin per ml on 7 separate days showed good reproducibility with a coefficient of variation of 0 to 11.7% for the latex assay compared with 7.01 to 22.2% for RIA. Recovery of amikacin in spiked sera varied between 93 and 108% for the latex assay compared with 90 and 100% for RIA. Because the procedure involves a titer, the latex agglutination inhibition card test produces results which are categorized rather than results which are continuous. However, it is a rapid and specific method for determining amikacin levels in clinical specimens and is particularly useful when processing small numbers of specimens.

Latex agglutination inhibition card test for gentamicin assay: clinical evaluation and comparison with radioimmunoassay and bioassay.

Gentamicin levels were determined in 100 serum specimens by a new latex agglutination inhibition card test, a radioimmunoassay (RIA), and a bioassay. Correlation coefficients determined by linear regression analysis demonstrated that the levels obtained by the latex agglutination inhibition card test had a high degree of correlation with the RIA and could be performed much faster and more economically when processing small numbers of specimens. The bioassay had a slightly lower degree of correlation with both the RIA and the latex test and was adversely influenced by concurrently administered antibiotics which could not be eliminated by beta-lactamase. When measuring gentamicin concentrations above 2 micrograms/ml, the coefficient of variation was less than 14% for the latex agglutination assay compared with 15% for the bioassay and 12% for RIA. The latex agglutination inhibition card test is a rapid, accurate, specific, and reproducible method for monitoring gentamicin levels in patients and is particularly applicable for laboratories processing small numbers of specimens.

Prevalence and nature of hyperamylasemia in acute alcoholism.

Determination of serum amylase activity in 100 consecutive patients admitted to an alcohol detoxification unit revealed hyperamylasemia in 39 cases. Further clinical evaluation of 15 of the 39 alcoholic patients with hyperamylasemia was unremarkable except for bilateral enlargement of the parotid glands in two cases. Nine of the 15 patients demonstrated markedly low amylase to creatinine clearance ratio; however, macroamylase complexes were not detected in the sera of any patients. Serum isoamylase separation revealed that the mean salivary isoamylase for the 15 alcoholic patients was significantly (P less than 0.05) elevated as compared to the control values. Individually, the salivary-type isoamylase was clearly elevated in ten patients while pancreatic type isoamylase was elevated in four. These data indicate that elevated serum amylase activity occurs frequently in alcoholic patients. Hyperamylasemia in a large number of alcoholic patients is nonpancreatic in origin and may be related to the injurious effect of ethanol on salivary glands and other tissues.

Evaluation of automated glucose oxidase methods for serum glucose: comparison to hexokinase of a colorimetric and an electrometric method.

Two automated glucose oxidase methods have been evaluated with respect to accuracy, precision, recovery, linearity and various potential interferences. Trinder's method on an AutoAnalyzer II had a between-day coefficient of variation (C.V.) of 2.6% (mean 228 mg/dl), was linear to 500 mg/dl, and produced a mean recovery of 99.7%. Comparison of Trinder's method with a manual, blanked hexokinase method yielded the regression equation: TR=--1.95 + HEX (1.04); Spearman's rho correlation coefficient was: 0.974. The Beckman System I glucose method had a between-day C.V. of 1.6% (mean 198 mg/dl), was linear to 500 mg/dl, and recovered an average of 98.0% of added glucose. Comparison with the same hexokinase method yielded: SYI = 1.27 + HEX (1.02: Spearman's rho = 0.991. None of the possible interfering compounds tested caused significant deviation of results by either method within the range of concentrations encountered physiologically. Trinder's method on the AutoAnalyzer II and the System I method are accurate, precise methods and are highly recommended for routine use in the clinical laboratory.