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Background: Liquid chromatography high-resolution mass spectrometry (LC-HRMS) with untargeted data collection is especially attractive for general unknown drug screening owing to its ability to identify unexpected compounds. LC-HRMS offers several advantages over traditional selected reaction monitoring (SRM) techniques and could be an ideal screening platform as long as its analytical performance is comparable to that of SRM-based methods., Methods: We developed a broad-spectrum drug screen on a high-resolution mass spectrometer [tandem quadrupole time-of-flight (QqTOF)] that collected data in an untargeted manner and compared its performance to a nominal mass instrument [triple quadrupole linear ion trap (QqLIT)] that collected data in a targeted manner. Both methods used information-dependent acquisition of product ion spectra. We evaluated the lower limits of detection and matrix effects for each method and compared their ability to identify drugs in 100 routine clinical urine samples. Additional information (patient prescription history, drug screening results, etc.) was used to confirm discordant results., Results: QqLIT was slightly more analytically sensitive than QqTOF; however, this difference did not significantly affect compound identification in patient samples. QqLIT identified 596 drugs in the urine samples, of which 531 (89%) were confirmed. QqTOF identified 515 drugs, of which 500 (97%) were confirmed. There were 562 instances of a confirmed drug (68 unique drugs) in the 100 urine samples; the methods were concordant in 469 of these instances., Conclusions: Overall, QqTOF performed similarly to QqLIT and could serve as an alternative method for general unknown screening., (© 2015 American Association for Clinical Chemistry.)

Background: The challenge in systematic toxicological analysis using gas chromatography and/or liquid chromatography coupled to mass spectrometry is to identify compounds of interest from background noise. The large amount of spectral information collected in one full-scan MS run demands the use of automated evaluation of recorded data files. We evaluated the applicability of the freeware deconvolution software AMDIS (Automated Mass Spectral Deconvolution and Identification System) for GC-MS-based systematic toxicological analysis in urine for increasing the speed of evaluation and automating the daily routine workload., Methods: We prepared a set of 111 urine samples for GC-MS analysis by acidic hydrolysis, liquid-liquid extraction, and acetylation. After analysis, the resulting data files were evaluated manually by an experienced toxicologist and automatically using AMDIS with deconvolution and identification settings previously optimized for this type of analysis. The results by manual and AMDIS evaluation were then compared., Results: The deconvolution settings for the AMDIS evaluation were successfully optimized to obtain the highest possible number of components. Identification settings were evaluated and chosen for a compromise between most identified targets and general number of hits. With the use of these optimized settings, AMDIS-based data analysis was comparable or even superior to manual evaluation and reduced by half the overall analysis time., Conclusions: AMDIS proved to be a reliable and powerful tool for daily routine and emergency toxicology. Nevertheless, AMDIS can identify only targets present in the user-defined target library and may therefore not indicate unknown compounds that might be relevant in clinical and forensic toxicology.

Background: In clinical and forensic toxicology, general unknown screening is used to detect and identify exogenous compounds. In this study, we aimed to develop a comprehensive general unknown screening method based on liquid chromatography coupled with a hybrid triple-quadrupole linear ion trap mass spectrometer., Methods: After solid-phase extraction, separation was performed using gradient reversed-phase chromatography. The mass spectrometer was operated in the information-dependent acquisition mode, switching between a survey scan acquired in the Enhanced Mass Spectrometry mode with dynamic subtraction of background noise and a dependent scan obtained in the enhanced product ion scan mode. The complete cycle time was 1.36 s. A library of 1000 enhanced product ion-tandem mass spectrometry spectra in positive mode and 250 in negative mode, generated using 3 alternated collision tensions during each scan, was created by injecting pure solutions of drugs and toxic compounds., Results: Comparison with HPLC-diode array detection and gas chromatography-mass spectrometry for the analysis of 36 clinical samples showed that linear ion trap tandem mass spectrometry could identify most of the compounds (94% of the total). Some compounds were detected only by 1 of the other 2 techniques. Specific clinical cases highlighted the advantages and limitations of the method., Conclusion: A unique combination of new operating modes provided by hybrid triple-quadrupole linear ion trap mass spectrometers and new software features allowed development of a comprehensive and efficient method for the general unknown screening of drugs and toxic compounds in blood or urine.

I have adapted, for use with a centrifugal analyzer, reagents from 10 different EMIT (Syva Co.) enzyme immunoassay urine drug-detection kits, reagents from another five serum kits, and reagents for determining ethanol. Assays for individual drugs in 100 microL of urine and in 100 microL of serum are performed simultaneously rather than sequentially in less than 15 min, with one-tenth the usual amount of reactants (sample + reagents). These assays, when combined with other easily performed tests, provide a rapid toxicology screen well-suited for evaluating acute emergency patients, and for performing a small number of daily routine screens. The total screen requires 500 microL of urine and 500 microL of serum. I summarize my experience with this procedure in its first year of use, and discuss differences between testing for the care of patients compared with searching for evidence of illegal drug use.

The equipment, methods, logistics, and results of doping-control analyses for the 1984 Los Angeles Olympic Games are discussed in this article. Within 15 days, 1510 different urine specimens underwent 9440 screening analyses by a combination of gas chromatography, gas chromatography-mass spectrometry, "high-performance" liquid chromatography, and radioimmunoassay. These tests covered more than 200 different drugs and metabolites, including psychomotor stimulants, sympathomimetic amines, central nervous system stimulants, narcotic analgesics, and anabolic steroids. The results are summarized by class of drug. Less than 2% of the samples were found to contain a banned drug.

In a prospective drug-testing program, individuals whose job classifications have been defined as sensitive are placed in a selection pool. Periodically, individuals are chosen from this pool for drug testing. Random selection is a fair and impartial approach for this because each individual has an equal chance of being selected. Random selection also serves to reinforce the deterrent potential of a drug-testing program because individuals in the pool never know when or if they will be selected. This uncertainty discourages drug usage because of the risk involved in being selected and tested. A random selection process generates a Poisson distribution of probabilities that can be used to predict how many times an individual will be selected during a specific time interval. This information can be used to model the selection part of a drug-testing program to determine whether specific conditions of testing are met. For example, the probability of being selected a given number of times during the testing period can be minimized or maximized by varying the frequency of the sampling process. Consequently, the Poisson distribution and the mathematics governing it can be used to structure a drug-testing program to meet the needs of any given situation.

A rapid extraction method preliminary to analysis for drugs in urine is described. Extraction columns containing purified cotton fiber (JETUBE, Manhattan Instrument Co., Santa Monica, Calif.) are shown to give high (approximately 90-97%) extraction efficiencies for some commonly prescribed or abused drugs, notably phenobarbital, amphetamine, morphine, and methadone. Analysis time, extraction efficiency, convenience, and eluate purity with a modified JET procedure are shown to be superior to results obtained with XAD-2 resin columns.

Confirmation of presumptive positive urine drug screens, necessary to minimize the reporting of false-positive results, can be costly and time-consuming. The predictive value model can be used to select the confirming tests and to calculate the confidence of the result. The predictive value of a test result is the probability, based on the sensitivity and specificity of the test, that the result is a true positive or a true negative. The predictive value model applied to toxicology screening tests for drugs of abuse showed that prevalence, in addition to sensitivity and specificity, was the factor controlling the confidence level of a result. For example, the predictive value of a positive result for a screening test that has a sensitivity of 99% and a specificity of 99%, applied to screening in a population with a prevalence of 1% is 0.50; for a prevalence of 10%, it is 0.92. Confirmation with a second, chemically independent, test of equal sensitivity and specificity increases the predictive value to 0.99.

A convenient screening procedure for presence of drugs of abuse in urine consists of two steps: adsorption of the drugs from urine onto a paper loaded with cation-exchange resin and detection of the adsorbed drugs by direct radioimmunoassay. The first step can be performed in the field, the second in a central laboratory. Storage and transport to the laboratory are simplified because specimens adsorbed on dried paper are stable and can be sent in letter-mail. In the laboratory, a small disc of the ion-exchange paper is exposed to antigen and antibody, rinsed, and tested for radioactivity. Discs treated with positive urines are more radioactive than discs from negative urines.