The secret life of crime labs.

Houston TX experienced a widely known failure of its police forensic laboratory. This gave rise to the Houston Forensic Science Center (HFSC) as a separate entity to provide forensic services to the City of Houston. HFSC is a very large forensic laboratory and has made significant progress at remediating the past failures and improving public trust in forensic testing. HFSC has a large and robust blind testing program, which has provided many insights into the challenges forensic laboratories face. HFSC's journey from a notoriously failed lab to a model also gives perspective to the resource challenges faced by all labs in the country. Challenges for labs include the pervasive reality of poor-quality evidence. Also that forensic laboratories are necessarily part of a much wider system of interdependent functions in criminal justice making blind testing something in which all parts have a role. This interconnectedness also highlights the need for an array of oversight and regulatory frameworks to function properly. The major essential databases in forensics need to be a part of blind testing programs and work is needed to ensure that the results from these databases are indeed producing correct results and those results are being correctly used. Last, laboratory reports of "inconclusive" results are a significant challenge for laboratories and the system to better understand when these results are appropriate, necessary and most importantly correctly used by the rest of the system.

Interference of the novel designer benzodiazepine 4'-chloro deschloroalprazolam with alprazolam analysis in toxicology and seized drug DUID casework.

In recent years, the emergence of the novel designer benzodiazepine 4'-chloro deschloroalprazolam has presented a new challenge for forensic laboratories by interfering with the identification and quantitation of alprazolam. As an isomer of alprazolam, 4'-chloro deschloroalprazolam has similar physicochemical properties and can be misidentified in casework samples as alprazolam without a specific method to differentiate the two analytes. Starting in late 2021, the Houston Forensic Science Center (HFSC) received toxicological and seized drug evidence indicating the presence of 4'-chloro deschloroalprazolam. An interference study was performed to supplement the laboratory's validated benzodiazepines method for toxicological samples to differentiate alprazolam from 4'-chloro deschloroalprazolam. This study showed that while the isomers could not be chromatographically resolved using the current method, they could be differentiated based on their retention times relative to the internal standard, alprazolam-d5. Based on these findings, the HFSC toxicology laboratory reports test results as "unsuitable for analysis due to an interference" if a suspected alprazolam peak elutes before the alprazolam-d5 peak, even if all identification and quantification criteria (e.g., retention time) were acceptable. Additionally, the seized drug and toxicology laboratories re-evaluated previously analyzed alprazolam-positive casework to determine if suspected 4'-chloro deschloroalprazolam had been misidentified as alprazolam. This report presents three cases: one case with toxicological evidence indicating the presence of both 4'-chloro deschloroalprazolam and alprazolam, and two cases with both seized drug material and toxicology evidence indicating the presence of 4'chloro deschloroalprazolam with no detected alprazolam.

Evaluation of suspected drug-facilitated sexual assault cases in the city of Houston from 2014 to 2020.

Drug-facilitated sexual assault is a form of sexual violence against an individual incapacitated by alcohol and/or drugs consumed voluntarily or covertly administered. The purpose of this study was to evaluate toxicological results and the associated demographics of sexual assault-related cases submitted to Houston Forensic Science Center from 2014 to 2020. In total, 1240 samples (1230 cases) were tested during the six-year period that consisted of blood, urine, or both specimens. Blood was analyzed for ethanol by dual-column headspace gas chromatography with flame ionization detection. Drug screen analysis was performed preferably on urine specimens using enzyme-linked immunosorbent assay. Positive screening results were confirmed upon request only due to laboratory policy. A total of 22% (n = 176) of requested samples were confirmed positive. Ethanol was the most prevalent substance detected, present in 17% of the samples (n = 212), followed by 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH) (n = 118). The combination of ethanol and THC-COOH was the most frequent one found (n = 17) in cases positive for two or more drugs (n = 101). Demographic data showed the majority of DFSA victims were white (25%) females (72%) with an average age of 27 years old (n = 348). Almost 90% of cases where the presence of drugs was confirmed resulted in no charges being made, either due to lack of suspect information or unknown reasons by the laboratory.

A Comparison of High Drug Concentrations in Impaired Driving and Postmortem Casework in Harris County, TX.

Drug-impaired driving is a growing public safety issue. Addressing impairment due to drugs other than ethanol can be challenging for forensic toxicologists as many factors need to be considered including the type of drug(s), drug-drug interaction, the dose(s) and the individual's physiological condition and drug use history. Interpretation of blood drug test results is additionally difficult as drug concentrations in impaired driving cases may overlap levels typically viewed as toxic. This study compares blood concentrations of drugs in impaired driving cases to those in postmortem cases in Houston, TX, from 2014 to 2020. Blood drug concentrations from driving while intoxicated (DWI) or driving under the influence of drugs (DUID) cases submitted to Houston Forensic Science Center (HFSC) and Harris County Institute of Forensic Sciences (HCIFS) were compared to postmortem blood test results from HCIFS. Eight DWI/DUID cases had drugs that exceeded impaired driving concentrations reported in the literature. These drugs included fentanyl (220 ng/mL), oxycodone (680 ng/mL), hydrocodone (310 and 490 ng/mL), clonazepam (330 ng/mL), methamphetamine (3,500 and 7,100 ng/mL) and tetrahydrocannabinol (THC) (160 ng/mL). For oxycodone and hydrocodone, the presented DWI/DUID cases exceeded 91% and 96% of postmortem concentrations, respectively. The 7,100 ng/mL methamphetamine DWI/DUID result was greater than 98% of postmortem cases. The presented DWI/DUID concentrations were higher than all but one postmortem case for clonazepam and higher than all postmortem cases for THC. This study demonstrates that extremely high drug concentrations in DWI/DUID casework blur the line between therapeutic/recreational and toxic concentrations.

Blind testing in firearms: Preliminary results from a blind quality control program.

Open proficiency tests meet accreditation requirements and measure examiner competence but may not represent actual casework. In December 2015, the Houston Forensic Science Center began a blind quality control program in firearms examination. Mock cases are created to mimic routine casework so that examiners are unaware they are being tested. Once the blind case is assigned to an examiner, the evidence undergoes microscopic examination and comparison to determine whether the fired evidence submitted was fired in the same firearm. Fifty-one firearms blind cases resulting in 570 analysis and comparison determinations were reported between December 2015 and June 2021. No unsatisfactory results were obtained; however, 40.3% of comparisons in which the ground truth was either elimination or identification resulted in inconclusive conclusions. Due to the quality of some of the evidence submitted, inconclusive results were not unexpected. A ground truth of elimination and comparison result of inconclusive was observed at a rate of 74%, while a ground truth of identification and comparison result of inconclusive was observed at a rate of 31%. Bullets (61.8%) were the main contributors to inconclusive conclusions; variables such as the assigned examiners, training program, examiner experience, and the intended complexity of the case did not significantly contribute to the results. The program demonstrates that the quality management system and firearms section procedures can obtain accurate and reliable results and provides examiners added confidence in court. Additionally, the program can be tailored to target specific research questions and provide opportunities for collaboration with other laboratories and researchers.

Everything Is Bigger in Texas: Alcohol Impaired Driving in Houston (2014-2018).

Driving while intoxicated (DWI) or driving under the influence of drugs (DUID) poses a continued public safety risk in Texas, which has one of the highest alcohol impaired traffic fatality rates. This study aimed to identify alcohol and drug use trends seen in DWI/DUID cases in the city of Houston from 2014 to 2018 to better understand the populations at risk. Blood samples submitted to the Houston Forensic Science Center (HFSC) were evaluated for blood alcohol concentration (BAC), drug concentrations and demographics. During the 5-year period, 12,682 Houston driver blood samples had a mean (median) BAC of 0.151 g/dL (0.167 g/dL) and age of 36.3 (34) years. Fifty percent of samples were white males. Seventy-five percent of samples were individuals aged 21 to 44 years. Between 2014 and 2018, the number of cases submitted nearly doubled, from approximately 2,000 cases per year to over 3,700. In 2014, 23% of cases submitted required further analysis per HFSC testing policy (drug screen and confirmation for DWI/DUID cases when BAC < 0.100 g/dL), which rose to 27% by 2018. Of those, 36% were polydrug cases, requiring two or more confirmation tests. Cannabinoids was the most common drug class detected (34% of cases analyzed for drugs), followed by benzodiazepines (25%), phencyclidine (20%), cocaine/metabolites (15%) and opioids (12%). Phencyclidine rose from the fifth-most commonly detected drug in 2014 to the second-most drug in 2018. Only 3% of all cases (n = 408) were negative for both alcohol and drugs. Communication between law enforcement and laboratory management is recommended to address growing caseload more effectively. The study limitations (e.g., limited scope of analysis) suggest the present data underestimated the full extent of impaired driving in Houston, indicating even more urgent needs for increasing resources and efforts to reduce this serious threat to public safety.

Houston Cocktail: Driving under influence of hydrocodone, alprazolam, and carisoprodol.

Concurrent use of opioids, benzodiazepines, and skeletal muscle relaxants potentiates the drug effect and respiratory depression via interactions of µ-opioid and GABAA receptors. In the early 2000s when abuse of prescription drugs began to spike, a potent combination including hydrocodone, alprazolam, and carisoprodol, aka the "Houston Cocktail" or "Holy Trinity", emerged that may give users heroin-like euphoria. This research evaluated driving while intoxicated (DWI) cases that tested positive for hydrocodone, alprazolam, and carisoprodol, between 2015 and 2019. The blood samples were collected from drivers and submitted by the Houston Police Department (HPD). They were subsequently analyzed for alcohol and drugs by reference laboratories or Houston Forensic Science Center (HFSC). Toxicological findings, demographic information, and observed impairment were evaluated for the Houston Cocktail-positive DWI cases. A total of 80 DWI/DUID cases positive for hydrocodone, alprazolam, and carisoprodol in blood in which the traffic offense occurred between May 2015 and December 2019 were identified. Among these Houston Cocktail cases, the mean (median, range) concentrations were 75 (61, 6.9-322) ng/mL for hydrocodone, 58 (48, 5.8-180) ng/mL for alprazolam, and 3.9 (3.0, 0.3-14; n = 68) µg/mL for carisoprodol; 80 (100%) and 23 (29%) cases were also positive for meprobamate (mean 13; range 1.2-41 µg/mL) and hydromorphone (1.8; 1.0-3.3 ng/mL), respectively; carisoprodol and meprobamate in 12 of the cases were qualitatively detected. Forty six percent of those cases were females and 54% were males; 44% were Blacks, 46% were Whites, and 10% were other races as identified by the arresting officer. Mean (median) age of the drivers was 36 (34) years, ranged from 22 to 60 years. Twenty eight percent of the cases were positive for the Houston Cocktail only; 21% had one other drug/metabolite, 28% two, 14% three, and 10% had four or more additional drugs/metabolites. Of the 80 cases, cannabinoids were the most frequently detected analytes (35%), followed by codeine (11%). The drivers exhibited driving problems related to lane position, vigilance, judgment, speed, and/or braking. Many of the drivers (70-84%) had red/glassy eyes, slurred speed, poor balance, HGN and impaired divided attention. The present study showed that despite a traffic safety risk, drivers in Houston continue to use this dangerous drug combination. The risk is further exacerbated by the fact that the many drivers had yet other drugs in the system besides the three drugs.

Toxicological and Demographic Profiles of Phencyclidine-Impaired Driving Cases in Houston.

Phencyclidine (PCP) was first synthesized in 1926 and originally developed in 1950s as a general anesthetic agent. Abuse of PCP declined at the national level since its first illicit use in 1960s, but it continues in certain areas including Houston. This research evaluates PCP-positive cases of driving while intoxicated (DWI) in 2013-2018. The blood samples were collected from drivers, submitted by the Houston Police Department and analyzed for alcohol and drugs. Toxicological findings and demographic information were evaluated for the impaired driving cases tested positive for PCP in blood. Additionally, the Drug Recognition Expert (DRE) findings were examined for 12 cases in 2018. A total of 615 DWI cases positive for PCP in blood were identified in which the traffic offense occurred between August 2013 and December 2018. The mean (median, range) PCP concentration was 47 (43, 7-180) ng/mL. A total of 23% of those cases were females, and 77% were males; 85% were blacks, 10% were whites and 5% were other races/ethnicities as identified by the arresting officer. The mean age was 37 years. No significant differences in median and distribution of PCP concentrations (P's > 0.05) were observed across the offense years and among demographic cohorts. A total of 43% of the cases were positive for PCP only. Among the remaining 57%, cannabinoids are the most common concurrently detected analytes (35%), followed by cocaine/metabolite (14%) and ethanol (13%). The proportion of black male PCP-positive drivers decreased in younger age groups. Common indications observed by DRE officers included slurred speech, chemical breath odor, watery and/or bloodshot eyes, vertical/horizontal gaze nystagmus and impaired coordination/balance. This study provides valuable regional information to better understand the demographic patterns of PCP-impaired drivers in Houston, TX over 6 years. The findings may aid in designing and implementing regulations and prevention programs to reduce PCP-impaired driving.

Implementation of a Blind Quality Control Program in a Forensic Laboratory.

A blind quality control (QC) program was successfully developed and implemented in the Toxicology, Seized Drugs, Firearms, Latent Prints (Processing and Comparison), Forensic Biology, and Multimedia (Digital and Audio/Video) sections at the Houston Forensic Science Center (HFSC). The program was put into practice based on recommendations set forth in the 2009 National Academy of Sciences report and is conducted in addition to accreditation required annual proficiency tests. The blind QC program allows HFSC to test its entire quality management system and provides a real-time assessment of the laboratory's proficiency. To ensure the blind QC cases mimicked real casework, the workflow for each forensic discipline and their evidence submission processes were assessed prior to implementation. Samples are created and submitted by the HFSC Quality Division to whom the expected answer is known. Results from 2015 to 2018 show that of the 973 blind samples submitted, 901 were completed, and only 51 were discovered by analysts as being blind QC cases. Implementation data suggests that this type of program can be employed at other forensic laboratories.

Implementation of a Blind Quality Control Program in Blood Alcohol Analysis.

Declared proficiency tests are limited in their use for testing the performance of the entire system, because analysts are aware that they are being tested. A blind quality control (BQC) is intended to appear as a real case to the analyst to remove any intentional or subconscious bias. A BQC program allows a real-time assessment of the laboratory's policies and procedures and monitors reliability of casework. In September 2015, the Houston Forensic Science Center (HFSC) began a BQC program in blood alcohol analysis. Between September 2015 and July 2018, HFSC submitted 317 blind cases: 89 negative samples and 228 positive samples at five target concentrations (0.08, 0.15, 0.16, 0.20 and 0.25 g/100 mL; theoretical targets). These blood samples were analyzed by a headspace gas chromatograph interfaced with dual-flame ionization detectors (HS-GC-FID). All negative samples produced `no ethanol detected' results. The mean (range) of reported blood alcohol concentrations (BACs) for the aforementioned target concentrations was 0.075 (0.073-0.078), 0.144 (0.140-0.148), 0.157 (0.155-0.160), 0.195 (0.192-0.200) and 0.249 (0.242-0.258) g/100 mL, respectively. The average BAC percent differences from the target for the positive blind cases ranged from -0.4 to -6.3%, within our uncertainty of measurement (8.95-9.18%). The rate of alcohol evaporation/degradation was determined negligible. A multiple linear regression analysis was performed to compare the % difference in BAC among five target concentrations, eight analysts, three HS-GC-FID instruments and two pipettes. The variables other than target concentrations showed no significant difference (P > 0.2). While the 0.08 g/100 mL target showed a significantly larger % difference than higher target concentrations (0.15-0.25 g/100 mL), the % differences among the higher targets were not concentration-dependent. Despite difficulties like gaining buy-in from stakeholders and mimicking evidence samples, the implementation of a BQC program has improved processes, shown methods are reliable and added confidence to staff's testimony in court.

External contamination of hair with cocaine: evaluation of external cocaine contamination and development of performance-testing materials.

The National Laboratory Certification Program undertook an evaluation of the dynamics of external contamination of hair with cocaine (COC) while developing performance testing materials for Federal Drug-Free Workplace Programs. This characterization was necessary to develop performance materials that could evaluate the efficacy of hair testing industry's decontamination procedures. Hair locks (blonde to dark brown/black) from five different individuals were contaminated with cocaine HCl. Hair locks were then treated with a synthetic sweat solution and hygienic treatments to model real-life conditions. Hair locks were shampooed daily (Monday through Friday) for 10 weeks, and samples of the hair locks were analyzed for COC, benzoylecgonine (BE), cocaethylene (CE), and norcocaine (NCOC). Three commercial analytical laboratories analyzed samples under three protocols: no decontamination procedure, individual laboratory decontamination, or decontamination by an extended buffer procedure at RTI International. Results indicated substantial and persistent association of all four compounds with all hair types. Hair that was not decontaminated had significantly greater quantities of COC and BE than did hair that was decontaminated. The only hair samples below detection limits for all four compounds were those decontaminated 1 h after contamination. Additionally, BE/COC ratios increased significantly over the 10-week study (regardless of decontamination treatment). From 21 days postcontamination until the end of the study, the mean BE/COC ratio for all hair types exceeded 0.05, the proposed Federal Mandatory Guidelines requirement. The largest variability in results was observed for samples decontaminated by participant laboratories. This suggests that current laboratory decontamination strategies will increase variability of performance testing sample results. None of the decontamination strategies used in the study were effective at removing all contamination, and some of the contaminated hair in this study would have been reported as positive for cocaine use based on the proposed Federal Mandatory Guidelines.

Urine benzodiazepine screening using Roche Online KIMS immunoassay with beta-glucuronidase hydrolysis and confirmation by gas chromatography- mass spectrometry.

Performance of the Roche Online KIMS (kinetic interaction of microparticles in solution) benzodiazepine (BZD) immunoassay (IA) with and without beta-glucuronidase treatment was evaluated on a Hitachi Modular automated IA analyzer calibrated using nordiazepam at 100 ng/mL. Reproducibility, linearity, accuracy, sensitivity, and interferences were evaluated. Precision of the assay (percent coefficient of variation (%CV)) with and without addition of the enzyme was less than 6% and 9%, respectively, with linearity (r(2) value of 0.9578 and 0.9746), respectively. Between-run precision of a 125 ng/mL nordiazepam control (n = 287) over 67 days, produced a %CV of 13.6% for the hydrolytic assay. Modification of the BZD assay to include automated hydrolysis of urinary BZD glucuronide conjugates was evaluated using three glucuronidated BZD standards prepared at concentrations ranging from 250 to 10,000 ng/mL. With hydrolysis, temazepam, oxazepam, and lorazepam glucuronides, produced cross-reactivities of 25%, 15%, and 20%, respectively. Without hydrolysis, the glucuronidated BZD standards produced less than 1% cross-reactivity in the assay. The ability of the assay to differentiate between positive and negative samples was evaluated by assaying 20 negative urine samples and serial dilutions of certified drug-free urine fortified with 28 different BZDs. All of the negative and positive urine samples produced the appropriate screening result. Cross-reactivities of 27 different BZDs, calculated as the normalized IA response divided by the BZD concentration that produced a response approximately equivalent to the response of a 100 ng/mL nordiazepam standard and multiplied by 100, ranged from 15% to 149%. Human urine samples (n = 28) that were previously found to contain BZDs by gas chromatography-mass spectrometry (GC-MS) also produced a positive BZD IA result. The IA was challenged with 78 potentially interfering compounds, and none produced a positive BZD response. As a part of the validation, a large number of human urine samples (29,500) were assayed using the modified Online BZD IA method to evaluate the performance of the method in production. Of the 29,500 samples tested, 80 produced a positive IA result. Analysis by GC-MS confirmed the presence of at least 1 BZD compound in 61 of the samples corresponding to a confirmation rate of 76%. The Online BZD IA modified by the automatic addition of beta-glucuronidase appears well adapted for the rapid detection of BZDs and their metabolites in human urine.

Comparison and evaluation of DRI methamphetamine, DRI ecstasy, Abuscreen ONLINE amphetamine, and a modified Abuscreen ONLINE amphetamine screening immunoassays for the detection of amphetamine (AMP), methamphetamine (MTH), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethamphetamine (MDMA) in human urine.

The performances of four immunoassays (DRI amphetamines, DRI ecstasy, Abuscreen ONLINE amphetamines, and a modified Abuscreen ONLINE amphetamines) were evaluated for control failure rates, sensitivity, and specificity for amphetamine (AMP), methamphetamine (MTH), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethamphetamine (MDMA). The two DRI reagents and the ONLINE reagents were run according to manufacturer specifications using a Roche Hitachi Modular DDP system. The modified ONLINE reagent was calibrated with MDMA and had 16mM sodium periodate added to the R2 reagent. These assays were run on approximately 27,500 human urine samples and 7000 control urine samples prepared at 350 and 674 ng/mL over the course of 8 days. All assays were calibrated using a single point, qualitative cutoff standard with the manufacturer-recommended compound at the Department of Defense cutoff (500 ng/mL). Gas chromatography-mass spectrometry (GC-MS) confirmation was conducted on screened-positive samples. Control performance for the manufacturer recommended assays was excellent, with a maximum qualitative control failure rate of 2.03%. The modified ONLINE reagent demonstrated poor control performance with a maximum failure rate of 38.3% and showed no improved MDMA sensitivity when compared with the ONLINE reagent; the confirmation rate (20%) was improved when compared with the production ONLINE reagent (8%). The DRI ecstasy reagent provided improved sensitivity for MDMA as compared with the ONLINE reagent, with approximately 23% more samples screening and confirming positive for MDMA and a confirmation rate of approximately 90%. The DRI methamphetamine reagent had a low confirmation rate (6% or less) and produced numerous positives for samples with only ephedrine or pseudoephedrine present.

Rapid simultaneous determination of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, and 3,4-methylenedioxyethylamphetamine in urine by solid-phase extraction and GC-MS: a method optimized for high-volume laboratories.

To facilitate analysis of high sample volumes, an extraction, derivatization and gas chromatographic-mass spectrometric analysis method was developed to simultaneously determine amphetamine (AMP), methamphetamine (MAMP), 3,4-methylenedioxyamphetamine (MDA) 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA) in urine. This method utilized a positive-pressure manifold cation-exchange polymer-based solid-phase extraction followed by elution directly into automated liquid sampler (ALS) vials. Rapid derivatization was accomplished using heptafluorobutyric anhydride (HFBA). Recoveries averaged 90% or greater for each of the compounds. Limits of detection were 62.5 ng/mL (AMP and MDEA), 15.6 ng/mL (MAMP), and 31.3 ng/mL (MDA and MDMA) using a 2-mL sample volume. The method was linear to 5000 ng/mL for all compounds using MDMA-d5 and MAMP-d14 as internal standards. Over 200 human urine samples previously determined to contain the target analytes were analyzed using the method. Excellent agreement was seen with previous quantitations. The method was challenged with 75 potentially interfering compounds and no interferences were seen. These interfering compounds included ephedrine, pseudoephedrine, phenylpropanolamine, and phenethylamine. The method resulted in dramatic reductions in processing time and waste production.

Occupational cocaine exposure of crime laboratory personnel preparing training aids for a military working dog program.

The potential for passive cocaine exposure was evaluated in crime laboratory employees preparing training aids for a military working dog program (MWD). The primary goal of the study was to elucidate the routes of exposure and implement procedural changes that would minimize this risk. Several work environments and laboratory procedures were examined by monitoring personal breathing zones (PBZ), ambient airborne cocaine levels in the laboratory spaces, and urinary levels of the primary cocaine metabolite, benzoylecgonine. The study was performed initially using current laboratory procedures to establish a baseline and to identify potential sources of exposure. A subsequent study was performed to determine the effectiveness of the follow-up procedure in reducing exposure. As a result of the changes, the 8-h time weighted averages (TWAs) were 40 to 80% lower in the follow-up study as compared to the baseline assessment. Dermal absorption and PBZ inhalation of cocaine during manufacture were likely the most significant source of cocaine exposure. Ambient airborne cocaine may have also contributed to the total exposure, but for most observations, the concentrations were significantly less than those determined from PBZ monitoring. The maximum ambient cocaine concentration was 0.0144 mg/m(3) compared to a maximum of 0.4004 mg/m(3) observed during PBZ monitoring. Occupational exposure decreased in the follow-up study because of the proper use of personal protective equipment and improvements in engineering controls.

LC-MS analysis of 2-oxo-3-hydroxy LSD from urine using a Speedisk positive-pressure processor with Cerex PolyChrom CLIN II columns.

A rapid and sensitive solid-phase extraction method for the lysergic acid diethylamide (LSD) metabolite 2-oxo-3-hydroxy-LSD (O-H-LSD) was developed for use in a forensic laboratory. The method uses a positive-pressure manifold anion-exchange polymer-based solid-phase extraction with subsequent analysis by liquid chromatography-mass spectrometry (LC-MS). The average extraction efficiency was 92%. The limits of detection (LOD) and quantitation (LOQ) were calculated at 250 pg/mL. The assay was linear, precise, and accurate from 250 to 30,000 pg/mL with an r(2) of 0.999. Samples including 93 compounds with properties similar to O-H-LSD, common over-the-counter products, prescription drugs and some of their metabolites, and other drugs of abuse were analyzed and produced no significant interference. This method has increased our efficiency in analyzing O-H-LSD by reducing the overall extraction and analysis time. The increase in extraction efficiency enabled decreased assay LOD and LOQ values while lowering the volume required for injection.

LC-mS analysis of human urine specimens for 2-oxo-3-hydroxy LSD: method validation for potential interferants and stability study of 2-oxo-3-hydroxy LSD under various storage conditions.

2-Oxo-3-hydroxy lysergic acid diethylamide (O-H-LSD), a major LSD metabolite, has previously been demonstrated to be a superior marker for identifying LSD use compared with the parent drug, LSD. Specifically, O-H-LSD analyzed using liquid chromatography-mass spectrometry has been reported to be present in urine at concentrations 16 to 43 times greater than LSD. To further support forensic application of this procedure, the specificity of the assay was assessed using compounds that have structural and chemical properties similar to O-H-LSD, common over-the-counter products, prescription drugs and some of their metabolites, and other drugs of abuse. Of the wide range of compounds studied, none were found to interfere with the detection of O-H-LSD or the internal standard 2-oxo-3-hydroxy lysergic acid methyl propylamide. The stability of O-H-LSD was investigated from 0 to 9 days at various temperatures, pH conditions, and exposures to fluorescent light. Additionally, the effect of long-term frozen storage and pH was investigated from 0 to 60 days. There was no significant loss of O-H-LSD under both refrigerated and frozen conditions within the normal human physiological pH range of urine (4.6-8.4). However, significant loss of O-H-LSD was observed in samples prepared at pH 4.6-8.4 and stored at room temperature or higher (24-50 degrees C).

Evaluation of a solid-phase extraction method for benzoylecgonine urine analysis in a high-throughput forensic urine drug-testing laboratory.

A novel extraction and derivatization procedure for the cocaine metabolite benzoylecgonine (BZE) was developed and evaluated for use in a high-volume forensic urine analysis laboratory. Extractions utilized a Speedisk 48 positive pressure extraction manifold and polymer-based cation-exchange extraction columns. Samples were derivatized by the addition of pentafluoropropionic anhydride and pentafluoropropanol. All analyses were performed in selected ion monitoring mode; ions included m/z 421, 300, 272, 429, and 303 with m/z 421 to 429 ratio used for quantitation. The average extraction efficiency was 80%. Seventy-five common over-the-counter products, including prescription drugs, drug metabolites, and other drugs of abuse, demonstrated no significant interference with respect to chromatography or quantitation. The limit of detection and limit of quantitation were calculated at 12.5 ng/mL, and the assay was linear from 12.5 to 20,000 ng/mL with an r2 of 0.99932. A series of 20 precision samples (100 ng/mL) produced an average response of 97.8 ng/mL and a percent coefficient of variation of 4.1%. A set of 79 archived human urine samples that had previously been found to contain BZE were analyzed by 3 separate laboratories. The results did not differ significantly from prior quantitation or between laboratories. The Speedisk has proven viable for a high-volume production facility reducing overall cost of analysis by decreasing analysis time and minimizing waste production while meeting strict forensic requirements.

Comparison of EMIT II, CEDIA, and DPC RIA assays for the detection of lysergic acid diethylamide in forensic urine samples.

In an effort to determine a practical, efficient, and economical alternative for the use of a radioimmunoassay (RIA) for the detection of lysergic acid diethylamide (LSD) in human urine, the performance of two photometric immunoassays (Dade Behring EMIT II and Microgenics CEDIA) and the Diagnostics Products Corp. (DPC) RIA were compared. Precision, accuracy, and linearity of the 3 assays were determined by testing 60 replicates (10 for RIA) at 5 different concentrations below and above the 500-pg/mL LSD cut-off. The CEDIA and RIA exhibited better accuracy and precision than the EMIT II immunoassay. In contrast, the EMIT II and CEDIA demonstrated superior linearity r2 = 0.9809 and 0.9540, respectively, as compared with the RIA (r2 = 0.9062). The specificity of the three assays was assessed using compounds that have structural and chemical properties similar to LSD, common over-the-counter products, prescription drugs and some of their metabolites, and other drugs of abuse. Of the 144 compounds studied, the EMIT II cross-reacted with twice as many compounds as did the CEDIA and RIA. Specificity was also assessed in 221 forensic human urine specimens that previously screened positive for LSD by the EMIT II assay. Of these, only 11 tested positive by CEDIA, and 3 were positive by RIA. This indicated a comparable specificity performance between CEDIA and RIA. This also was consistent with a previously reported high false-positive rate of EMIT II (low specificity). Each of the immunoassays correctly identified LSD in 23 out of 24 human urine specimens that had previously been found to contain LSD by gas chromatography-mass spectrometry at a cut-off concentration of 200 pg/mL. The CEDIA exhibited superior precision, accuracy, and decreased cross-reactivity to compounds other than LSD as compared with the EMIT II assay and does not necessitate the handling of radioactive materials.