尿液与血液标本中HIV-1抗体的检测结果分析

目的 比较应用ELISA法检测尿液和血液标本中HIV-1抗体结果的一致性.方法 对15244例被检测尿液和血液HIV-1抗体者进行回顾性分析,应用ELISA方法分别采用尿和血初筛试剂检测尿液和血液标本中HIV-1抗体.结果 15244例被检测的样本中,26份样本尿和血标本检测结果同时为阳性,以血液标本为标准,检测尿液HIV-1抗体的灵敏度为100%,特异性为99.33%,尿液标本和血液标本检测的符合率达99.25%.结论 可通过尿液ELISA初筛诊断试剂进行HIV-1感染情况的监测和筛查.     

Detection of morphine in blood and urine samples from horses administered poppy seeds and morphine sulfate orally

The objective of this study was to determine if the administration of poppy seeds to horses would result in detectable concentrations of morphine in urine and blood samples, as has been shown to occur in humans. In this study blood and urine samples were collected following administration of poppy seeds and morphine sulfate orally to four horses. Urine samples were subjected to enzyme-linked immunosorbent assay (ELISA) for the presence of morphine. All urine samples testing positive by ELISA, as well as plasma samples collected after administration of the 10-g doses of poppy seeds, were analyzed by gas chromatography-mass spectrometry for the presence of morphine. Morphine was detectable in the plasma samples for at least 4 h after administration of 10 g of poppy seeds. Morphine was detectable in urine samples for up to 24 h after administration of 10 g, 5 g, and 1 g of poppy seeds and 426.7 microg of morphine as morphine sulfate. The results of this study indicate that horses that consume or are administered poppy seeds may have detectable concentrations of morphine in their urine and plasma for hours after administration.     

A comparison of a commercial microdiffusion method and gas chromatography for ethanol analysis

A simple, rapid ethanol screen based on a microdiffusion technique (Toxi-Lab alcohol test) was evaluated using whole blood and urine samples and compared to a direct injection gas chromatographic technique. A total of 204 samples were analyzed, 124 whole blood specimens and 80 urine samples. The whole blood samples showed agreement in 79 of 80 samples that tested positive by gas chromatography. All 44 whole blood samples that tested negative by gas chromatography were also negative by the microdiffusion method. All urine samples, both positive and negative, as well as all control specimens showed agreement. The one whole blood sample that did not agree had an ethanol level by gas chromatography below the stated detection limit (0.01%) of the microdiffusion method. The microdiffusion method was found to be a viable screening test for the presence of ethanol in whole blood and urine. It is relatively inexpensive and rapid and is therefore an ideal method for screening samples before a more expensive or time-consuming confirmatory method.     

The isomeric metabolites of doxepin in equine serum and urine

Due to its tranquilizing properties, the tricyclic antidepressant doxepin may be misused as a doping agent in competition horses. Therefore, efficient analytical procedures are required to detect this drug in samples submitted for doping control. To screen for parent doxepin in equine blood and urine, a less specific method has been accepted employing gas chromatography (GC) combined with electron impact (EI) mass spectrometry (MS). The aim of this study was identification of doxepin metabolites providing more specific MS data to verify positives resulting from screening. Thus, after a horse was given doxepin-HCl (1 mg/kg, i.v.), blood and urine were analyzed for free or conjugated metabolites using GC combined with EI- and positive chemical ionization (PCI) MS. In both of the sample materials, cis- and trans-isomers of desmethyldoxepin were detected for up to 48 h after treatment using trifluoracetylation and GC/EI-MS. Following enzymic hydrolysis of urine and propionylation of extracts, each four isomers of hydroxy desmethyldoxepin and hydroxydoxepin were recovered for up to 24 and 48 h, respectively. These compounds were characterized by their EI- and PCI-mass spectra. Although distinct positions of the hydroxyl groups could not be determined, the presence of each two cis/trans-isomeric pairs of differently monohydroxylated metabolites may be assumed. Results reported here suggest, that screening horses for parent doxepin should be completed by analysis of its major isomeric metabolites, desmethyldoxepin and hydroxydoxepin, providing MS data specific enough for confirmatory analysis.     

Drug population in one thousand geographically distributed urine specimens

Hundreds of drugs and other substances are excreted in urine. In a comprehensive drug screen, it is impossible to identify every detectable substance. In order to delineate the most commonly detected drugs, approximately 1000 urine specimens from geographically distributed clinical laboratories were analyzed for drug substances. Twenty-nine (29) clinical laboratories submitted up to 76 consecutive urine specimens found to be positive for substances other than nicotine and caffeine. Specimens were analyzed by thin layer chromatography and special procedures for salicylates, benzoylecgonine, PCP, benzodiazepines, and cannabinoid metabolites. Every specimen was also tested for opiates, PCP, and cannabinoid metabolites by an enzyme immunoassay procedure. The total number of drugs detected in 1000 specimens was 3014, an average of three drugs per specimen; and 110 different drugs were identified. Of these, 50 drugs accounted for 95% of the total detected; 21% of the specimens contained cannabinoid metabolites, and 4% of the specimens contained cocaine and/or benzoylecgonine. Most of the specimens were routine, and only 4% originated from comatose patients.     

Disposition and metabolic profile of the weak androgen Dehydroepiandrosterone (DHEA) following administration as part of a nutritional supplement to exercised horses

In order to ensure the welfare of performance horses and riders as well as the integrity of the sport, the use of both therapeutic and illegal agents in horse racing is tightly regulated. While Dehydroepiandrosterone (DHEA) is not specifically banned from administration to racehorses in the United States and no screening limit or threshold concentration exists, the metabolic conversion of DHEA to testosterone make its presence in nutritional supplements a regulatory concern. The recommended regulatory threshold for total testosterone in urine is 55 and 20 ng/mL for mares and geldings, respectively. In plasma, screening and confirmation limits for free testosterone (mares and geldings), of no greater than 0.1 and 0.025 ng/mL, respectively are recommended. DHEA was administered orally, as part of a nutritional supplement, to 8 exercised female thoroughbred horses and plasma and urine samples collected at pre‐determined times post administration. Using liquid chromatography‐mass spectrometry (LC‐MS), plasma and urine samples were analyzed for DHEA, DHEA‐sulfate, testosterone, testosterone‐sulfate, pregnenolone, androstenedione, and androstenediol. DHEA was rapidly absorbed with maximal plasma concentrations reaching 52.0 ± 43.8 ng/mL and 32.1 ± 12.9 ng/mL for DHEA and DHEA sulfate, respectively. Free testosterone was not detected in plasma or urine samples at any time. Maximum sulfate conjugated testosterone plasma concentrations were 0.98 ± 1.09 ng/mL. Plasma testosterone‐sulfate concentrations did not fall below 0.1 ng/mL and urine testosterone‐sulfate below 55 ng/mL until 24–36 h post DHEA administration. Urine testosterone sulfate concentrations remained slightly above baseline levels at 48 h for most of the horses studied. Copyright © 2014 John Wiley & Sons, Ltd.     

A comprehensive study on the variations in urinary concentrations of endogenous gamma-hydroxybutyrate (GHB)

This study was designed to supplement previous attempts to establish an accurate range of normal endogenous gamma-hydroxybutyrate (GHB) concentrations in random antemortem urine samples. Furthermore, its purpose was to ascertain the effect of gender, race, age, medications, and select medical conditions on endogenous concentrations of GHB in urine and the proposed endogenous urinary GHB cutoff of 10 microg/mL. Urine samples (n = 207) were provided by subjects who reported that they had never used GHB. As part of the collection process, subjects also completed a short survey to collect information about gender, race, age, orally ingested medications, and select medical conditions. All specimens were analyzed in duplicate for the presence of endogenous GHB using a previously reported headspace gas chromatography-mass spectrometry method. The data were analyzed for tendencies among different population groups. GHB concentrations ranged from 0.00 to 2.70 microg/mL in all specimens, with a median concentration of 0.24 microg/mL. Males (n = 130) had an average endogenous GHB concentration of 0.27 microg/mL (0.00-2.70 microg/mL), whereas females (n = 77) averaged 0.29 microg/mL (0.00-0.98 microg/mL). Select medical conditions and participants' race, age ranges, and medications that were used within 48 h prior to collection were also evaluated. We believe this to be the most comprehensive study on endogenous GHB concentrations in urine to date. The results of this study will aid the interpretation of low GHB concentrations measured in urine samples, particularly in investigations of drug-facilitated crimes.     

唾液皮质醇与血浆皮质醇、尿游离皮质醇测定的相关性分析及其临床价值

目的探讨正常人唾液皮质醇、血浆皮质醇和24 h尿游离皮质醇三者之间的相关性。方法选择30例健康受试者同时检测唾液皮质醇、血浆皮质醇和24 h尿游离皮质醇。男子组和女子组各15例。采用化学发光技术测定,唾液标本采用唾液收集器收集,血标本为血浆,24 h尿液标本用硼酸防腐。结果唾液皮质醇和血浆、24 h尿液皮质醇在正常男女组之间均无显著性差异（P＆gt;0.05）,唾液皮质醇与血浆皮质醇（r=0.91）、唾液皮质醇与24 h尿游离皮质醇（r=0.95）,均存在显著的相关性。结论唾液皮质醇可作为库欣氏综合征的筛查指标。     

Pharmacokinetics of dextromethorphan and its metabolites in horses following a single oral administration

Dextromethorphan is an N‐methyl‐D‐aspartate (NMDA) non‐competitive antagonist commonly used in human medicine as an antitussive. Dextromethorphan is metabolized in humans by cytochrome P450 2D6 into dextrorphan, which is reported to be more potent than the parent compound. The goal of this study is to describe the metabolism of and determine the pharmacokinetics of dextromethorphan and its major metabolites following oral administration to horses. A total of 23 horses received a single oral dose of 2 mg/kg. Blood samples were collected at time 0 and at various times up to 96 h post drug administration. Urine samples were collected from 12 horses up to 120 h post administration. Plasma and urine samples were analyzed using liquid chromatography‐mass spectrometry, and the resulting data analyzed using non‐compartmental analysis. The C_max, T_max, and the t_1/2 of dextromethorphan were 519.4 ng/mL, 0.55 h, and 12.4 h respectively. The area under the curve of dextromethorphan, free dextrorphan, and conjugated dextrorphan were 563.8, 2.19, and 6,691 h*ng/mL respectively. In addition to free and glucuronidated dextrorphan, several additional glucuronide metabolites were identified in plasma, including hydroxyl‐desmethyl dextrorphan, desmethyl dextrorphan, and three forms of hydroxylated dextrorphan. Dextromethorphan was found to be eliminated from the urine predominately as the O‐demethylated metabolite, dextrorphan. Several additional metabolites including several novel hydroxy‐dextrorphan metabolites were also detected in the urine in both free and glucuronidated forms. No significant undesirable behavioural effects were noted throughout the duration of the study. Copyright © 2016 John Wiley & Sons, Ltd.     

Fatality due to acute alpha-methyltryptamine intoxication

In February 2003, the Miami-Dade County Medical Examiner Department reported the first known death in the country related to alpha-methyltryptamine (AMT). AMT is an indole analogue of amphetamine investigated in the 1960s as an antidepressant, stimulant, and monoamine oxidase inhibitor. Today, AMT is recognized as a powerful psychedelic drug among high school and college-aged men and women. Its popularity is partly due to the multitude of anecdotal websites discussing AMT as well as its legality and availability for purchase via the Internet prior to April 2003. Emergency designation of AMT as a Schedule 1 controlled substance by the Drug Enforcement Administration occurred shortly after the death in Miami-Dade County. The case in Miami involved a young college student who, prior to death, advised his roommate that he was "taking hallucinating drugs" and as a result had "discovered the secret of the universe". Approximately 12 h later, the roommate discovered the deceased lying in bed unresponsive. An empty 1-g vial of AMT was recovered from the scene and sent to the toxicology laboratory. Initial screening of urine by enzyme-multiplied immunoassay technique was positive for amphetamines, and the basic drug blood screen detected a small peak later identified by mass spectrometry as AMT. For quantitation, AMT was isolated using solid-phase extraction, derivatized with pentafluoropropionic anhydride, and analyzed using gas chromatography-mass spectrometry. Quantitative analysis was based upon m/z 276, 303, and 466 for AMT and m/z 306, 333, and 496 for the internal standard, 5-methoxy-alpha-methyltryptamine. A linear calibration curve from 50 to 500 ng/mL was used to calculate the concentration of AMT in the samples and controls. Blood, tissue, and gastric specimens were diluted to bring the observed concentration within the limits of the standard curve. Matrix matched controls were extracted and analyzed with each run. Postmortem iliac vein blood revealed 2.0 mg/L, gastric contents (48 g collected at autopsy) contained 9.6 mg total of AMT, liver contained 24.7 mg/kg, and the brain contained 7.8 mg/kg. An additional Medical Examiner case from another jurisdiction revealed 1.5 mg/L in antemortem serum.     

Findings of ten years of clinical drug screening

During the years from 1977 to 1986, 21,410 specimens were collected for clinical drug screening from 13,589 patients. The specimens included 9,530 blood samples, 9,184 urine samples and 2,717 samples of gastric contents. One or more drugs/drug metabolites were detected in 74% of the specimens. Gastric contents were positive most often (85%) followed by urine (79%) and blood (66%). Although the 5 most frequently detected drugs did not change (caffeine, ethanol, nicotine, salicylates and acetaminophen), the presence of cocaine, phenylpropanolamine, diphenhydramine and the group of cyclic antidepressants steadily increased over the time period. Over 200 different drugs/drug metabolites were detected in the drug screens, contributing a significant amount of information for the appropriate diagnosis and treatment of the patients for which the testing was requested.     

Detection of myo‐inositol tris pyrophosphate (ITPP) in equine following an administration of ITPP

Myo‐Inositol tris pyrophosphate (ITPP) is a powerful allosteric modulator of haemoglobin that increases oxygen‐releasing capacity of red blood cells. It is capable of crossing the red blood cell membrane unlike its open polyphosphate analog myo‐inositol hexakisphosphate (IHP). Systemic administration of ITPP enhanced the exercise capacity in mice. There have been rumours of its abuse in the horse racing industry to enhance the performance of racing horses. In this paper, the detection of ITPP in equine plasma and urine after an administration of ITPP is reported. A Standardbred mare was administered 200 mg of ITPP intravenously. Urine and plasma samples were collected up to 120 h post administration and analyzed for ITPP by liquid chromatography‐tandem mass spectrometry. ITPP was detected in post administration plasma samples up to 6 hours. The peak concentration was detected at 5 min post administration. In urine, ITPP was detected up to 24 h post administration. The peak concentration was detected at 1.5 h post administration. Copyright © Her Majesty the Queen in Right of Canada 2013.     

Disposition of isoflupredone acetate in plasma,urine and synovial fluid following intra‐articular administration to exercised Thoroughbred horses

The use of isoflupredone acetate in performance horses and the scarcity of published pharmacokinetic data necessitate further study. The objective of the current study was to describe the plasma pharmacokinetics of isoflupredone acetate as well as time‐related urine and synovial fluid concentrations following intra‐articular administration to horses. Twelve racing‐fit adult Thoroughbred horses received a single intra‐articular administration (8 mg) of isoflupredone acetate into the right antebrachiocarpal joint. Blood, urine and synovial fluid samples were collected prior to and at various times up to 28 days post drug administration. All samples were analyzed using liquid chromatography‐Mass Spectrometry. Plasma data were analyzed using a population pharmacokinetic compartmental model. Maximum measured plasma isoflupredone concentrations were 1.76 ± 0.526 ng/mL at 4.0 ± 1.31 h and 1.63 ± 0.243 ng/mL at 4.75 ± 0.5 h, respectively, for horses that had synovial fluid collected and for those that did not. The plasma beta half‐life was 24.2 h. Isoflupredone concentrations were below the limit of detection in all horses by 48 h and 7 days in plasma and urine, respectively. Isoflupredone was detected in the right antebrachiocarpal and middle carpal joints for 8.38 ± 5.21 and 2.38 ± 0.52 days, respectively. Results of this study provide information that can be used to regulate the use of intra‐articular isoflupredone in the horse. Copyright © 2015 John Wiley & Sons, Ltd.     

Effectiveness of saliva and fingerprints as alternative specimens to urine and blood in forensic drug testing

In forensic drug testing, it is important to immediately take biological specimens from suspects and victims to prove their drug intake. We evaluated the effectiveness of saliva and fingerprints as alternative specimens to urine and blood in terms of ease of sampling, drug detection sensitivity, and drug detection periods for each specimen type. After four commercially available pharmaceutical products were administered to healthy subjects, each in a single dose, their urine, blood, saliva, and fingerprints were taken at predetermined sampling times over approximately four weeks. Fourteen analytes (the administered drugs and their main metabolites) were extracted from each specimen using simple pretreatments, such as dilution and deproteinization, and were analyzed using liquid chromatography/mass spectrometry (LC/MS). Most of the analytes were detected in saliva and fingerprints, as well as in urine and blood. The time‐courses of drug concentrations were similar between urine and fingerprints, and between blood and saliva. Compared to the other compounds, the acidic compounds, for example ibuprofen, acetylsalicylic acid, were more difficult to detect in all specimens. Acetaminophen, dihydrocodeine, and methylephedrine were detected in fingerprints at later sampling times than in urine. However, a relationship between the drug structures and their detection periods in each specimen was not found. Saliva and fingerprints could be easily sampled on site without using special techniques or facilities. In addition, fingerprints could be immediately analyzed after simple and rapid treatment. In cases where it would be difficult to immediately obtain urine and blood, saliva and fingerprints could be effective alternative specimens for drug testing. Copyright © 2015 John Wiley & Sons, Ltd.     

Characterization of bromhexine and ambroxol in equine urine: effect of furosemide on identification and confirmation

The purpose of this study was two-fold: (1) to develop a simple and sensitive screening procedure for identifying and confirming bromhexine and ambroxol and, (2) to determine the effect of furosemide on the detection of bromhexine, ambroxol, or their metabolites in urine. Female horses (450-550 kg) treated with bromhexine or ambroxol (1 g, p.o.) were used. Urine samples were collected up to 48 h post-drug administration and analysed. Blind samples were used in evaluating the sensitivity of these methods and reproducibility of the results. Bromhexine and ambroxol were extensively metabolized in the horse. These agents and their respective metabolites were identified and confirmed using thin-layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS), respectively. Hydroxy-bromhexine and desmethyl-bromhexine were major metabolites found to be unique to bromhexine-treated horses. These metabolites selectively absent from ambroxol-treated horse urine provide a chemical means to distinguish bromhexine from ambroxol administration in horses. These specific metabolites were similarly identified and confirmed in "blind" horse urine samples. The concomitant presence of furosemide (300 mg, i.v.) with bromhexine or ambroxol did not mask the presence of these agents or alter their metabolite profile. By application of the methods described in this study, bromhexine and ambroxol metabolites in horse urine can be easily identified and confirmed.     

An automated alcohol dehydrogenase method for ethanol quantification in urine and whole blood

In a previous work, an automated alcohol dehydrogenase method for the quantification of ethanol in whole blood (blood) specimens was presented. In the present work the application of the method to urine specimens has been investigated. Also, method robustness to routine analysis of urine and blood specimens during a period of eight months is shown. The limits of detection and quantification for urine were 0.0012 g/dL and 0.0042 g/dL, respectively. Relative standard deviations for the repeatability and within-laboratory reproducibility were in the ranges 1.4-4.1% and 1.8-4.6%, respectively. The method was compared with two headspace gas chromatography-flame ionization detection methods using authentic forensic urine specimens (n = 305) and blood specimens (n = 3186). Passing-Bablok regression for the concentration range 0.01-0.48 g/dL (urine) and 0.002-0.40 g/dL (blood) showed a statistically significant difference, for urine y = 0.9313 (0.9250 - 0.9377)x + 0.0038 (0.0029-0.0044) and for blood y = 0.9493 (0.9491 - 0.9495)x + 0.0032 (0.00318-0.00323), at 95% confidence level. The results of the external quality control specimens were in accordance with the reported theoretical concentrations.     

Effects of caffeine on locomotor activity of horses: determination of the no-effect threshold

Caffeine is the legal stimulant consumed most extensively by the human world population and may be found eventually in the urine and/or blood of race horses. The fact that caffeine is in foods led us to determine the highest no-effect dose (HNED) of caffeine on the spontaneous locomotor activity of horses and then to quantify this substance in urine until it disappeared. We built two behavioural stalls equipped with juxtaposed photoelectric sensors that emit infrared beams that divide the stall into nine sectors in a 'tic-tac-toe' fashion. Each time a beam was interrupted by a leg of the horse, a pulse was generated; the pulses were counted at 5-min intervals and stored by a microcomputer. Environmental effects were minimized by installing exhaust fans producing white noise that obscured outside sounds. One-way observation windows prevented the animals from seeing outside. The sensors were turned on 45 min before drug administration (saline control or caffeine). The animals were observed for up to 8 h after i.v. administration of 2.0, 2.5, 3.0 or 5.0 mg caffeine kg(-1). The HNED of caffeine for stimulation of the spontaneous locomotor activity of horses was 2.0 mg kg(-1). The quantification of caffeine in urine and plasma samples was done by gradient HPLC with UV detection. The no-effect threshold should not be greater than 2.0 microg caffeine ml(-1) plasma or 5.0 microg caffeine ml(-1) urine.