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alpha-benzyl-N-methylphenethylamine (BNMPA), an impurity of illicit methamphetamine synthesis: III. Detection of BNMPA and metabolites in urine of methamphetamine users.

Eighty urine specimens collected from drug rehabilitation programs, which had been screened by immunoassay and confirmed positive by gas chromatography-mass spectrometry (GC-MS) for methamphetamine, were further analyzed for alpha-benzyl-N-methylphenethylamine (BNMPA) and its urinary metabolites, N-demethyl-BNMPA, diphenyl-2-propanone (DP2P), diphenyl-2-propanol, p-OH-N-demethyl-BNMPA, and p-OH-BNMPA. BNMPA is an impurity of illicit methamphetamine synthesis. Analysis of BNMPA and its metabolites was performed by quantitative GC-MS following beta-glucuronidase hydrolysis, liquid-liquid extraction, and derivatization with heptafluorobutyric anhydride. Two urine specimens contained detectable amounts of BNMPA and/or its metabolites. One contained trace amounts (greater than the limit of detection but less than the limit of quantitation) of N-demethyl-BNMPA and DP2P, as well as 0.04 mg/L p-OH-N-demethyl-BNMPA. The other contained trace amounts of BNMPA, p-OH-BNMPA, and p-OH-N-demethyl-BNMPA, as well as 0.03 mg/L N-demethyl-BNMPA. Prior to analyzing these urine specimens, pure reference material of p-OH-BNMPA was made available, and analysis confirmed our previous tentative identification of p-OH-BNMPA as a major metabolite of BNMPA. Detection of BNMPA or its metabolites in biological samples may serve as a marker of illicit methamphetamine administration.

Amphetamine↗

Uranium bioassay and radioactive dust measurements at some uranium processing sites in Egypt--health effects.

The safety of radiation workers in the uranium mining industry requires close and continuous monitoring of their working conditions. In this study, external radiation surveillance, radioactive dust monitoring and the bioassay of uranium were carried out in some processing sites. As dust represents one of the most important sources of radiation exposure in mills and mines, dust monitoring and bioassays were performed for a sample of workers on the production lines. The concentration of uranium in air ranged from 22.6 x 10(-7) to 11.1 x 10(-5) Bq cm-3, and the exposure levels ranged from 1 to 80 microSv h-1. Laser fluorimetric determination of uranium in urine samples showed concentrations in the range 8.4-29.2 micrograms L-1. Renal function parameters, such as serum creatinine and urea, and hematological parameters were determined in an attempt to correlate them with radiation exposure and the health status of the workers. Urine specimens collected from workers at the ore crushing and separation site showed elevated concentrations of uranium (up to 29.2 micrograms L-1) and a strong correlation between these concentrations and the registered serum creatinine. The mean uranium excretion in the investigated group was more than 20 times the occupational exposure decision level for urine uranium of 0.8 microgram L-1.

Adult↗

Experience with a urine opiate screening and confirmation cutoff of 2000 ng/mL.

Until recently, most laboratories used an opiate immunoassay screening and confirmation cutoff value of 300 ng/mL for codeine and morphine detection by gas chromatography-mass spectrometry (GC-MS). The cutoff value for opiates was increased to 2000 ng/mL or higher in various laboratories because of concerns that small doses of codeine and foods containing poppy seeds would give a positive opiate-screening result. Workplace drug-testing programs in the U.S. raised the opiate cutoff value to 2000 ng/mL on 30 November 1998. The objective of this study is to describe the results of opiate testing of 8600 urine specimens collected over 24 months with a 2000-ng/mL screening and confirmation (codeine and morphine) cutoff value. Specimens were screened by the EMITdau opiate assay using an in-house 2000-ng/mL morphine calibrator. Presumptive positive findings (N = 621) were analyzed quantitatively by GC-MS for codeine and morphine. One hundred and eighty six urine specimens were positive for codeine and morphine (> 2000 ng/mL), 298 specimens were positive for codeine only (> 2000 ng/mL) and 26 specimens were positive for morphine only (> 2000 ng/mL). All remaining specimens had codeine and morphine values < 2000 ng/mL. The codeine and morphine confirmation rate in this program reduced from 7.1% in 1994-1996 (300-ng/mL cutoff) to 2.1% in 1997-1998 with a 2000-ng/mL cutoff value. The codeine-only confirmation rate lowered from 6.6% (300-ng/mL cutoff) to 3.4% (2000-ng/mL cutoff). It was concluded that increasing opiate screening and codeine and morphine confirmation cutoff values led to > 300% reduction in the confirmed-positive rate for codeine and morphine and a 47% reduction in codeine-only confirmations in a urine drug-testing program where codeine was the major opiate used.

Codeine↗

Evaluation of urine as a clinical specimen for diagnosis of hepatitis a.

The present study pertains to the evaluation of urine as a specimen for detection of anti-hepatitis A virus (anti-HAV) antibodies. Immunoglobulin M (IgM), IgG, and IgA capture enzyme-linked immunosorbent assays for hepatitis A were performed on paired serum and urine specimens collected from hepatitis A patients (n = 92), healthy individuals (n = 100), non-A hepatitis patients (n = 70), and patients with nonhepatic diseases (n = 64, including 37 renal disease patients). Hepatitis A patients seropositive for anti-HAV IgM showed 95.65% uropositivity. No false-positive reactions were observed in control groups. The uropositivity of anti-HAV IgM persisted during the convalescent phase of the disease. Anti-HAV IgG uropositivity correlated well with corresponding seropositivity in all groups (P > 0.05 for each). No significant difference between the proportions of serum and urine positivity for anti-HAV IgA was noted (P > 0.05 for each). Using seroreactivity as a "gold standard," the sensitivity and specificity for anti-HAV IgM, anti-HAV IgG, and anti-HAV IgA tests with urine as a specimen were found to be 95.65 and 100%, 97.76 and 76.47%, and 92.23 and 88.18%, respectively. Urine appears to be comparable to serum for diagnosis of recent and past infection with hepatitis A.

Adolescent↗

Immunoassay for detection of cocaine/metabolites in oral fluids.

There is growing interest in the use of alternate biological fluids for drug testing. An advantage of oral fluids is that collection can be made from individuals under direct observation without undue embarrassment or invasion of privacy. This study evaluated the STC Cocaine Metabolite MICRO-PLATE EIA for use in detection of cocaine and metabolites in oral fluids. Intra- and interassay precision of the EIA was < 10%. The EIA was cross-reactive to benzoylecgonine (100%), cocaine (> 12.9%), and cocaethylene (13.8%), but did not demonstrate detectable cross-reactivity with other commonly encountered medicants. Evaluation of a series of potential adulterants of oral fluids indicated that common household chemicals and foodstuffs did not alter the outcome of EIA testing for cocaine metabolite. Analysis by EIA and by gas chromatography-mass spectrometry (GC-MS) of oral fluids and urine specimens collected from current drug users in treatment programs and subjects participating in research studies involving controlled dosing of cocaine provided assessment of the clinical sensitivity and specificity of the STC Cocaine Metabolite EIA. Analysis of the data by means of receiver operating characteristic (ROC) plot indicated that the optimal cutoff concentration for the oral fluids EIA was 10 ng/mL. In comparison to GC-MS (10-ng/mL combined cutoff concentration for cocaine and benzoyleogonine), the EIA (10-ng/mL cutoff concentration) demonstrated a sensitivity, specificity, and accuracy of 95%, 82%, and 88%, respectively. The oral fluids EIA was slightly less sensitive than the urine EIA (300-ng/mL cutoff concentration) for the detection of cocaine metabolite with a sensitivity, specificity, and accuracy of 73%, 85%, and 88%, respectively. Overall, testing of oral fluids for cocaine metabolite with the STC Cocaine Metabolite MICRO-PLATE EIA appears to offer a viable alternative to urine for detection of recent cocaine use.

Cocaine↗

Quantitative determination of circulating antigens in human schistosomiasis mansoni using an indirect hemagglutination assay.

In serum and urine specimens collected from a group of Schistosoma mansoni infected individuals from Makundju, Zaire, the schistosome circulating anodic antigen (CAA) and the circulating cathodic antigen (CCA) were quantitatively determined using an indirect hemagglutination reaction with sheep erythrocytes sensitized with mouse IgM monoclonal antibodies directed against these circulating antigens. Levels of CAA in serum (up to 5 ng/ml) and CCA in serum and urine (up to 50 ng/ml) were strongly correlated with egg excretion and with each other. No correlation was found between egg excretion and antibody levels against the circulating antigens. Antigen was detectable only in patients excreting greater than 500 eggs per gram of feces.

Animals↗

Antimicrobial activity in urine: effect on leukocyte count and bacterial culture results.

A total of 500 urine specimens collected from non-hospitalized patients were investigated for leukocyte count by microscopy, bacterial recovery by culture, and detection of antimicrobial activity by disc diffusion. Results indicated that 21% of the specimens contained leukocytes > or = 50/cu mm, whereas 14.4% of specimens revealed significant bacterial growth. The use of Staphylococcus aureus ATCC 29737 as a test bacterium detected the antimicrobial activity in 19.2% of the urine specimens, while the use of two clinical isolates of Escherichia coli detected this activity in fewer specimens. Evidence is provided which indicates that bacterial growth inhibitors in urine are mainly due to the intake of antibiotics by patients. These inhibitors confused the interpretation of urine culture results as they were found to play a major role in the occurrence of sterile pyorrhea and in reducing the number of specimens yielding significant bacterial growth.

Anti-Bacterial Agents↗

Biological variation in analyte concentrations in urine of apparently healthy men and women.

Analytical, intra-individual, and interindividual components of variation for sodium, potassium, urea, creatinine, calcium, and phosphate were estimated from results for 24-h urine specimens collected from 15 apparently healthy individuals every four weeks for 40 weeks. Expressed as output, mean values differed for men and women, except for calcium. Our data on intra-individual variation were similar to those obtained for 10 men by Shephard et al. (Clin Chem 1981;27:569-73). Calculated analytical goals are easily attained by current methods. Reference values for urine creatinine are useful only when expressed as output and stratified according to gender. The ratios of intra- to interindividual variation generally increase on such stratification; separate reference values for men and women are therefore required for analytes expressed as output. Measurements of sodium and potassium in urine should be reported as concentration, but output terms are favored for the other analytes. Differences for two serial results from an individual must be rather large to differ statistically.

Adult↗

Nephrotoxicity of cyclosporine A in the rat. II. Reversible changes in intranephronal and urinary catalytic activities of N-acetyl-beta-D-glucosaminidase.

Cyclosporine A (15, 30 or 50 mg/kg.day) or olive oil (30 mg/kg.day) were administered orally to 32 male Sprague-Dawley rats for 12 or 24 days, or withdrawn for 24 days following 24 days of treatment. The specific activity of a lysosomal marker enzyme, N-acetyl-beta-D-glucosaminidase, was determined fluorometrically in single nephron segments microdissected from lyophilized kidney sections of these animals and of an additional 2 normal rats. The segments were classified according to their normal or reduced succinate dehydrogenase activity as detected in stained adjacent sections. In addition, urine specimens collected after 12, 24, 36, and 48 days of the experiment were tested for N-acetyl-beta-D-glucosaminidase activity. After treatment with cyclosporine A, changes in the activities of N-acetyl-beta-D-glucosaminidase were found only in the proximal tubules. In the convoluted segments with normal succinate dehydrogenase activity, the activity of N-acetyl-beta-D-glucosaminidase was 138-163%, and in those with reduced succinate dehydrogenase activity, it was unchanged or 66-83% of the control values. In the straight segments with reduced succinate dehydrogenase activity, the activity of N-acetyl-beta-D-glucosaminidase increased gradually along the medullary rays (122-214%) to the outer stripe of the outer medulla (178-263%) in comparison to the control values. In the urine specimens, the activity of N-acetyl-beta-D-glucosaminidase was increased to 148-152%. These tubular and urinary changes were similar for each dosage and treatment period. After withdrawal of cyclosporine A they were not present. The variety of alterations occurring within the lysosomes along the proximal tubules of cyclosporine-A-treated rats implies the convoluted part as the site of increased release of N-acetyl-beta-D-glucosaminidase into the urine.

Acetylglucosaminidase↗

[Levels of metabolites of selected chemical compounds in the urine of workers in pitch coke plant].

In urine samples collected from pitch coke plant workers just before and after occupational exposure, differentiated concentrations of phenol (20.8-692.8 mg/dm3), p-cresol (51.8-590 mg/dm3), I-naphtol (4.9-63.7 mg/dm3) and benzo(a)pyrene (0.3-18.9 ug/dm3) have been found. The occurrence of enhanced concentrations of phenol in 10.1%, p-cresol in 31.9%, I-naphthol in 13.5% and benzo(a)pyrene in 72.8% of the test urine specimens collected prior to exposure points to a slow excretion of these metabolites of chemical compounds inherent in the work environment. Furthermore, the urinary level of the metabolites in particular coke plant workers points to different individual exposure.

Air Pollutants, Occupational↗

Rapid assessment of sexually transmitted diseases in a sentinel population in Thailand: prevalence of chlamydial infection, gonorrhoea, and syphilis among pregnant women--1996.

OBJECTIVE: To determine the prevalence of sexually transmitted diseases (STDs) among pregnant women in Thailand, where case reporting suggests a marked decrease in STDs following a campaign promoting condom use during commercial sex. DESIGN: Cross sectional study of women at their first visit to the study hospitals' antenatal clinics in Chiang Rai (n = 500) and Bangkok (n = 521). METHODS: First catch urine specimens were tested for Chlamydia trachomatis and Neisseria gonorrhoeae using the Amplicor CT/NG polymerase chain reaction assay. Syphilis and HIV serological testing were performed in the study hospitals' laboratories. RESULTS: The prevalence of chlamydial infection was 5.7%, gonorrhoea 0.2%, and syphilis 0.5% (all VDRL or RPR titres were < or = 1:4). The prevalence of HIV infection was 7.1% in Chiang Rai and 2.9% in Bangkok. In a multivariate logistic regression analysis, chlamydial infection was associated with younger age and with higher gestational age at first antenatal clinic visit, but was not associated with marital status, gravidity, city of enrollment, or HIV infection status. CONCLUSIONS: There was a low prevalence of gonorrhoea and syphilis among these pregnant women in Thailand. Chlamydial infection was detected at a higher prevalence, especially among younger women and women registering later for antenatal care. Testing of pregnant women using easily collected urine specimens and a sensitive nucleic acid amplification assay is a feasible method of rapidly assessing chlamydial and gonococcal prevalence.

Adolescent↗

Urinary excretion of 5-hydroxyindole-3-acetic acid and 5-hydroxytryptophol after oral loading with serotonin.

The urinary excretion patterns of the serotonin (5-hydroxytryptamine; 5-HT) metabolites 5-hydroxyindole-3-acetic acid (5-HIAA) and 5-hydroxytryptophol (5-HTOL) were examined after ingestion of bananas, a food rich in 5-HT. The bananas contained on an average 25 micrograms 5-HT/g pulp. Both urinary 5-HIAA and 5-HTOL increased markedly (15- to 30-fold) shortly after eating 3-4 bananas, with the highest concentrations found in urine specimens collected after 2-4 h, and did not return to normal until after 8-10 h. The excretion of 5-HIAA increased from a control mean value of 3.9 mg/24 h to 12.7 mg/24 h, when conventional diets were supplemented with 3-4 bananas. The corresponding results for 5-HTOL were 16.8 micrograms/24 h and 60.7 micrograms/24 h, respectively. Of the banana-derived 5-HT ingested, 60-80% was recovered in the urine as 5-HIAA and only 0.3-0.5% as 5-HTOL. However, since both the time-course and relative increase in 5-HTOL was similar to that of 5-HIAA, there was no effect on the urinary 5-HTOL to 5-HIAA ratio. By contrast, acute alcohol consumption produced a considerable elevation of this ratio.

Administration, Oral↗

Urinary screening for alprazolam and its major metabolites by the Abbott ADx and TDx analyzers with confirmation by GC/MS.

Alprazolam is a short-acting triazolobenzodiazepine with anxiolytic and antidepressant properties. It has a half-life of 10-15 hours after multiple oral doses. Approximately 20% of an oral dose is excreted unchanged in the urine. The major urinary metabolites are alpha-OH alprazolam glucuronide and 3-HMB benzophenone glucuronide. The objective of this study was to characterize the reactivity of alprazolam and three metabolites in the Abbott ADx and TDx urinary benzodiazepine assays compared with the EMIT d.a.u. benzodiazepine assay. Alprazolam (at 300 ng/mL) gave an equivalent response as the 300 ng/mL low control (nordiazepam). alpha-OH alprazolam gave an equivalent response to this control between 300-500 ng/mL and 4-OH alprazolam between 500-1000 ng/mL. The 3-HMB benzophenone was not positive even at 10,000 ng/mL. The ADx screening assay was positive in 26 of 31 urine specimens collected from alprazolam-treated patients. All 31 of these specimens were confirmed positive for alpha-OH alprazolam by GC/MS after enzymatic hydrolysis and formation of a TMS derivative. For the TDx, 27 of 31 specimens were positive for benzodiazepines and all 31 were confirmed by GC/MS. All 5 of the negative ADx specimens and 4 of 5 TDx specimens contained 150-400 ng/mL of alpha-OH alprazolam. In conclusion, both the ADx and TDx urine benzodiazepine assays are acceptable screening assays for alprazolam use when the alpha-OH alprazolam concentration is greater than 400 ng/mL.

Alprazolam↗

Glycosaminoglycans in urine, articular and periarticular tissues in streptozotocin diabetes in rats.

Experimental data support the hypothesis that insulin deficiency increases the degradation of glycosaminoglycans (GAGs) and decreases their synthesis. The purpose of this study was to investigate the effect of streptozotocin-induced experimental diabetes on mucopolysaccharide content of the articular and periarticular tissues as well as on the urine GAGs excretion. In two groups (A and B) of ten white Wistar rats, experimental diabetes was induced after administration of streptozotocin in a dose of 65 and 45 mg/kg of body weight, respectively. The animals of group A (short term diabetes) and B (long term diabetes) were sacrificed on the 8th and 87th day of the experimental procedure respectively. GAGs were determined in urine specimens collected before and on the 8th day in group A and before and on 3rd, 8th, 16th, 23rd, 36th, 52nd, 71st and the 87th day in group B. At the end of the experiment articular and periarticular tissue specimens were examined for changes in their mucopolysaccharides content and compared to a group of five animals of the same age and dietary conditions (control group, C). We found a marked increase (P less than 0.001) of urine GAGs excretion during streptozotocin-induced diabetes as compared to the respective values before the experiment in both groups, while in group B a positive relationship (P less than 0.001) between 24 hour urine GASGs excretion, and the duration of diabetes was observed. The histochemistry of the articular and periarticular tissues revealed marked decrease in GAGs content, as well as hardening of the tendons.

Animals↗

Stereoselective hydroxylation of tacrine in rats and humans.

An enantiospecific method was developed for assessing the stereochemistry of tacrine (9-amino-1,2,3,4-tetrahydroacridine monohydrochloride monohydrate; THA) metabolism to 1-hydroxytacrine (1-OH-THA) in humans and rats. In addition, limited metabolic studies with human liver microsomal preparations were conducted, and the stereochemistry of rac-1-OH-THA disposition was also examined. The analytical method incorporates an achiral normal phase separation and isolation of 1-OH-THA, followed by a chromatographic step using chiral normal-phase chromatography to resolve the enantiomers of 1-OH-THA. The achiral method was applied to quantitation of total 1-OH-THA in human urine specimens collected for 24 hr following administration of a single 40 mg oral dose of tacrine to 15 healthy elderly volunteers. Total 1-OH-THA accounted for approximately 5% of the administered dose. THA and 2-OH-THA were also quantitated and found to comprise < 1% and approximately 2% of the administered dose, respectively. 4-OH-THA was not detectable. The dextrorotatory (+)-isomer comprised approximately 94% of the 1-OH-THA recovered in urine. In vitro studies utilizing human liver microsomes found enantioselective formation of the (+)-isomer (approximately 90%), whereas incubations with rac-1-OH-THA showed residual substrate to be racemic. The method was also applied to determination of the enantiomeric composition of 1-OH-THA in the urine of rats given a single oral 16 mg/kg dose of THA. The percentage of 1-OH-THA excreted in urine as the (+)-isomer was 94%.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Detection of Salmonella spp. in clinical specimens by capture enzyme-linked immunosorbent assay.

A capture enzyme-linked immunosorbent assay (ELISA; Kirkegaard and Perry Laboratories, Gaithersburg, Md.) was used to detect Salmonella spp. in clinical and artificially inoculated specimens. In patients with bacteremia caused by Salmonella spp., 48% (12 of 25) and 82% (13 of 16) of serum and urine specimens, respectively, were positive for Salmonella spp., as determined by ELISA. All serum and urine specimens collected from healthy individuals (25 specimens) or patients whose blood cultures grew gram-negative bacteria other than Salmonella spp. (18 specimens) were negative for Salmonella spp., as determined by ELISA. For blood culture bottles in which Salmonella spp. (16 specimens) was grown the ELISA was positive (100%), while it was negative for all the 65 blood culture bottles in which gram-negative bacteria other than Salmonella spp. (42 specimens) or gram-positive bacteria (23 specimens) were grown. All samples of urine (16 specimens), stool (8 specimens), serum (16 specimens), culture media (12 specimens), and blood culture bottles (reported sterile after 2 weeks of incubation; 16 specimens) that were artificially inoculated with 10(3) to 10(7) CFU of four species of Salmonella per ml were positive by ELISA. Similar specimens inoculated with or containing various species other than Salmonella were negative by this test. Thus, ELISA offers a promising opportunity for the rapid detection of Salmonella spp. in clinical microbiology laboratories.

Antigens, Bacterial↗

Measured versus self-reported compliance with doxycycline therapy for chlamydia-associated syndromes: high therapeutic success rates despite poor compliance.

OBJECTIVE: To compare self-reported doxycycline compliance in men and women attending an STD clinic with indications for Chlamydia trachomatis treatment to compliance measured using microprocessor-containing medication vials to count each time and date medication vials were opened. A secondary objective was to correlate outcomes of therapy, as measured by symptom resolution and persistence of chlamydial nucleic acids, with measured doxycycline compliance. METHODS: Between September 1995 and July 1997, Medication Event Monitoring System (MEMS) caps were used to measure compliance with recommended doxycycline therapy (14 doses over 7 days) in patients treated for presumed C. trachomatis infections. Polymerase chain reaction (PCR) assays for C. trachomatis were performed on urine specimens collected at the time of follow-up evaluation. RESULTS: Of 221 evaluable participants, although 90% reported taking their medication as directed, only 33 (16%) managed this level of compliance according to data obtained from the MEMS cap. Although 144 (65%) patients took more than 11 of 14 doses over 8 days, 147 (67%) participants had at least one interval of 24 hours or longer between doses in an 8-day period. Of 81 participants with positive C. trachomatis cultures at enrollment, follow-up urine PCR for C. trachomatis was positive in 5 (6%). Medication Event Monitoring System data for four of the five patients with positive PCR tests as follow-up showed each had two or more 24-hour intervals when their medication vials were not opened and three of four had opened their vials less than 11 times. CONCLUSIONS: This study suggests that few patients take medications as prescribed and that self-report substantially underestimates medication noncompliance. Despite poor compliance, there were few treatment failures.

Adolescent↗

Influence of the transport on the outcome of the bacteriological analysis of dog urine comparison of three transport tubes.

A sterile plastic tube, a boric acid-glycerol-sodium formate tube and a dip-slide tube were compared for transport of dog urine for bacteriological examination under practice conditions, at ambient temperature. In a first part, 50 dog urine samples were cultured on agar plates and on a dip-slide kit within 2 h after collection and after storage at 20 degrees C in a sterile tube and in a boric acid tube for 24 h and 48 h. Culture of the samples stored in the boric acid tube and culture on the dip-slide yielded results which correlated very well with those of the culture of fresh urine. However, culture of the samples stored in the sterile tube yielded up to 65% false positive results. In a second part, a comparison was drawn with culture results of 100 dog urine specimens collected by different practitioners and simultaneously mailed to our laboratory in a sterile tube, in a boric acid tube and in a dip-slide kit. Samples sent in the boric acid and in the dip-slide tube showed comparable culture results. Culture of the samples sent in the plastic tube yielded 53% false positive results in comparison with those of the samples preserved in boric acid.

Animals↗