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In contrast to the numerous reports on the pharmacological effects of Δ(9)-tetrahydrocannabinol (THC), the pharmacological activity of another substituent of Cannabis sativa, cannabichromene (CBC) remains comparatively unknown. In the present study, we investigated whether CBC elicits cannabinoid activity in the tetrad assay, which consists of the following four endpoints: hypomotility, antinociception, catalepsy, and hypothermia. Because cannabinoids are well documented to possess anti-inflammatory properties, we examined CBC, THC, and combination of both phytocannabinoids in the lipopolysaccharide (LPS) paw edema assay. CBC elicited activity in the tetrad that was not blocked by the CB(1) receptor antagonist, rimonabant. Moreover, a behaviorally inactive dose of THC augmented the effects of CBC in the tetrad that was associated with an increase in THC brain concentrations. Both CBC and THC elicited dose-dependent anti-inflammatory effects in the LPS-induced paw edema model. The CB(2) receptor, SR144528 blocked the anti-edematous actions of THC, but not those produced by CBC. Isobolographic analysis revealed that the anti-edematous effects of these cannabinoids in combination were additive. Although CBC produced pharmacological effects, unlike THC, its underlying mechanism of action did not involve CB(1) or CB(2) receptors. In addition, there was evidence of a possible pharmacokinetic component in which CBC dose-dependently increased THC brain levels following an i.v. injection of 0.3mg/kg THC. In conclusion, CBC produced a subset of behavioral activity in the tetrad assay and reduced LPS-induced paw edema through a noncannabinoid receptor mechanism of action. These effects were augmented when CBC and THC were co-administered. [ABSTRACT FROM AUTHOR]

There has been an exponential surge in the presence and use of cannabinoids since the federal legalization of hemp (Agricultural Improvement Act of 2018). This growth is attributed to delta-9-tetrahydrocannabinol (delta-9-THC) and cannabidiol (CBD), the most abundant phytocannabinoid components of cannabis and hemp, respectively, but with many other emerging THC analogs. Structurally, these analogs are similar to delta-9-THC, yet very little information is available about their potency and even less information is available regarding their detectability using commercially available cannabinoid screening kits. Due to their structural similarity, current cannabinoid homogeneous immunoassay screening methods may be able to detect these emerging cannabinoid analogs and their metabolites. Six urine immunoassay kits (Abbott Cannabinoids-Abbott Diagnostics, LZI Cannabinoids (cTHC) Enzyme Immunoassay-Lin-Zhi International, DRI® Cannabinoid Assay and CEDIA™ THC-Thermo Fisher Scientific, ONLINE DAT Cannabinoid II-Roche Diagnostics and Syva EMIT®II Plus-Siemens Healthineers) were evaluated at two different cutoff concentrations: 50 ng/mL and 20 or 25 ng/mL, assay dependent. The analysis was performed on an Abbott Architect Plus c4000 (Abbott Diagnostics). Delta-8-THC, CBD, olivetol and their major metabolites, and delta-10-THC and HHC carboxylic acid chiral analogs were evaluated. The cross-reactivity was evaluated by preparing each analyte at 20, 50, 100 and 1,000 ng/mL in urine. Analytes that did not cross-react at 1,000 ng/mL for a cutoff were considered not detectable. If detected, the lowest concentration was used as the decision point to determine the precision at the immunoassay's cutoff. The six commercially available urine cannabinoid homogeneous immunoassay screening kits cross reacted with the following analogs: delta-8-THC, 11-OH-delta-8-THC, 11-COOH-delta-8-THC, 6-OH-CBD, 7-OH-CBD, all delta-10-THC and HHC carboxylic acid chiral analogs and olivetol with varying selectivity depending on the screening kit and cutoff concentration. The kits did not cross-react with the following analogs: CBD, 7-COOH-CBD, Abnormal CBD, CBDA-A and olivetolic acid., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

There has been a surge in the presence and use of cannabinoids since the federal legalization of hemp (Agricultural Improvement Act of 2018). This increase is attributed not only to the use of ∆9-tetrahydrocannabinol (∆9-THC) and cannabidiol, the most abundant phytocannabinoid components of cannabis and hemp, respectively, but also to the use of many other emerging THC analogs. Structurally, these analogs are similar to ∆9-THC. Urine specimens for drug analysis are often collected offsite and transported to a laboratory for analysis. Screening assays are usually the first step in urine drug testing. These assays are usually qualitative and automated, which for negative specimens, reduce cost and reporting time. The stability of ∆9-THC and its metabolites has been known for some time; however, the stability of emerging analogs has not been elucidated, and therefore, assuming equivalent storage stability can be erroneous. Previous work assessed the cross-reactivity of ∆8-THC and its major metabolites, the ∆10-THC chiral analogs and the chiral 11-COOH-hexahydrocannabinol analogs. Stability was assessed for each analyte at a concentration two times greater than the analytes' determined decision point. Samples were prepared in drug-free urine at three different pHs (4.5, 7 and 9) and stored at three different temperatures (4°C, 20°C and 45°C) in triplicate. Samples were analyzed utilizing the Lin-Zhi International Cannabinoids Enzyme Immunoassay cannabinoid screening kit calibrated at the 25 ng/mL cut-off. Overall, the cannabinoid analogs produced diminishing instrument responses depending on pH and temperature. The parent analogs were not detected after a single day at 45°C regardless of pH. In general, carboxylic acid analogs at the acidic pH (4.5) produced diminished instrument responses when compared to their counterparts stored at neutral (7) and basic (9) pH. The time, storage temperature and pH of urine specimens may affect the screening results of specimens collected for cannabinoid drug screening., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Bone health supplements containing strontium are available without prescription, however, the effects of strontium interference on clinical laboratory calcium measurement procedures are unknown., Methods: To evaluate strontium interference on total calcium measurements, plasma pools with exogenously added strontium were measured by 3 total calcium measurement procedures. For ionized calcium measurements, whole blood pools prepared with exogenously added strontium were measured by 2 ionized calcium measurement procedures. An inductively coupled plasma mass spectrometry assay (ICP-MS) was validated for research measurements of strontium content in commercially available supplements., Results: Exogenous strontium addition to plasma caused positive bias for total calcium measurements. Strontium concentrations of 1.0 mg/dL (0.114 mmol/L), 2.5 mg/dL (0.284 mmol/L), and 5.0 mg/Dl (0.568 mmol/L) resulted in mean biases of 1.9% to 3.5%, 4.9% to 9.0%, and 10.8% to 19.2%, respectively, for total calcium measurement procedures. Biases for ionized calcium measurements were less than 4.5% for a strontium concentration of 5.0 mg/dL (0.568 mmol/L). An in-house-developed ICP-MS assay for strontium in commercially available supplements exhibited within-laboratory and within-run coefficients of variation of less than 3%, and a linear response was obtained over the assay analytical measurement range of 10 to 100 000 ng/mL (0.0001 to 1.141 mmol/L). Strontium recovery for the ICP-MS assay was 97.1% to 105.3%. The largest amount of strontium measured in dietary supplements was 395 mg in a 1054 mg tablet., Conclusions: Some dietary supplements contain larger amounts of strontium than indicated on the product label. High concentrations of strontium may cause significant interference for total calcium measurement procedures, but ionized calcium measurement procedures are not significantly affected., (© American Association for Clinical Chemistry 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objectives: The ability to detect fentanyl analogs in urine aids in patient management. Little is published about the new ARK™ Fentanyl II Assay formulation's ability to detect fentanyl analogs. Norfentanyl (fentanyl metabolite) cross-reactivity with the ARK II assays is 7%, while the Immunalysis SEFRIA assay norfentanyl cross-reactivity is approximately 0.005%. The purpose of this study was to determine the new ARK II and SEFRIA fentanyl assays' detection of 58 fentanyl analogs., Design & Methods: Drug-free urine was fortified with 0-100 ng/mL (0-0.297 µmol/L) of the fentanyl analog and analyzed using the previously evaluated immunoassays. Results were compared to molecular structure. Of the 58 analogs tested at ≤ 100 ng/mL (0-0.297 µmol/L), the ARK II and SEFRIA assays produced 51 and 57 positive results respectively. The cross-reactivity of the assay was predominantly determined by the location of the modification. Most modifications to the aniline ring and/or amide group did not affect the ARK II or SEFRIA assay. Modifications to the piperidine ring decreased detection by ARK II assay. Of the 7 compounds which were undetected by the ARK II assay, all had modifications to the N-alkyl chain. Norsufentanil was not detected by either assay and was the only analog not detected by the SEFRIA assay., Conclusions: The ARK II and Immunalysis fentanyl immunoassays can detect a range of fentanyl analogs with acryl, butyryl, or furanyl modifications to the amide group or aniline ring of the molecule. N-alkyl chain and piperidine ring modifications significantly affect the ARK II assay's ability to detect the analogs, while the SEFRIA assay appeared less affected and detected all analogs tested except for norsufentanil, which was also not detected by the ARK II assay., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Inc.)

Background: We describe a case of symptomatic bradycardia resulting from ivabradine toxicity by measurement of ivabradine levels, of which there are limited reports in the literature., Case Presentation: A 43-year-old White female presented with several days of near syncope and dizziness accompanied by a drop in her heart rate to 50 beats per minute. She was taking ivabradine for inappropriate sinus tachycardia. After excluding several other causes of bradycardia, we made the diagnosis of ivabradine toxicity by measurement of serum ivabradine levels, an approach that is currently not clinically available., Conclusions: Measurement of serum ivabradine levels and knowledge of the pharmacokinetic properties of the drug can be utilized to confirm the diagnosis of ivabradine toxicity., (© 2022. The Author(s).)

Tramadol is an opioid used in the treatment of moderate to moderately severe pain. Tramadol's use during pregnancy is generally avoided and may cause some reversible withdrawal effects in neonates, and its use during lactation is not licensed by the manufacturer. A small clinical trial reported infants were exposed to <3% of a mother's tramadol dose through breast milk with no evidence of harmful effects. Presented is a case study of breast milk, neonatal urine, and neonatal oral fluid for the analysis of tramadol and its metabolites, along with the validation of a method for the analysis of tramadol, O-desmethyltramadol, and N-desmethyltramadol in breast milk. Tramadol and its metabolites were extracted by solid-phase extraction after saponification of breast milk to remove lipids. Samples were analyzed by ultra-pressure liquid chromatography-tandem mass spectrometry. To the author's knowledge, this is the first report of tramadol and its metabolites in neonatal oral fluid. The breast milk concentrations were 63, 22, and 76 ng/mL for the analysis of tramadol, O-desmethyltramadol, and N-desmethyltramadol, respectively, on day of life 12. On day of life 20, the breast milk concentrations were 1,254, 388, and 937 ng/mL for the analysis of tramadol, O-desmethyltramadol, and N-desmethyltramadol, respectively. Oral fluid concentrations were 1,011, 1,499, and 406 ng/mL for the analysis of tramadol, O-desmethyltramadol, and N-desmethyltramadol, respectively, on day of life 20. Oral fluid concentrations were similar to breast milk for tramadol, almost four times higher for O-desmethyltramadol, and less than half for N-desmethyltramadol. The absolute infant dose was calculated to be 10 μg/kg/day and 294 μg/kg/day for tramadol on day of life 12 and 20, respectively., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Introduction: The St. Ignatius bean of the Strychnos ignatii tree and Nux Vomica homeopathic products presumably could contain the toxic alkaloids strychnine and brucine. This study aimed to determine the amount of these toxic alkaloids in some commercially available Nux Vomica products and the St. Ignatius bean and to determine if overdose of these products could result in clinically significant toxicity., Methods: Using ultra-performance liquid chromatography-tandem mass spectrometry, various formulations of Nux Vomica products and St. Ignatius beans were analyzed for strychnine, and brucine with detection limits set at 0.1 ng/g., Results: None of the analyzed Nux Vomica products contained any detectable strychnine or brucine, while the expected strychnine dose from a St. Ignatius bean would be < 0.001 mg., Conclusions: Overall, our study reveals that the amount of strychnine in homeopathic Nux Vomica products or St. Ignatius beans are not likely to result in clinically significant strychnine toxicity., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Neonatal drug exposure is currently assessed using meconium, urine, blood, hair, or umbilical cord tissue/blood. Due to the invasiveness, challenges, and limitations of collection, and/or analytical difficulties of these matrices, oral fluid may be a more desirable matrix in diagnosing opioid exposure and risk for opioid withdrawal in neonatal abstinence syndrome. Traditional oral fluid collection devices are not viable options as they are too large for neonates' mouths and may contain chemicals on the collection pad. Unstimulated and stimulated infant oral fluid samples have been used for therapeutic drug monitoring as an alternative matrix to blood. The objective of this study was to assess the viability of a simple oral fluid collection system using a sterile foam-tipped swab rinsed in phosphate-buffered saline. Two infants were administered fentanyl for post-operative pain relief while hospitalized in the Neonatal Intensive Care Units at the Children's Hospital of Richmond of Virginia Commonwealth University. Oral fluid samples were collected at 16 h, 2 days, and/or 7 days following the start of intravenous infusion of fentanyl. Samples were analyzed by ultra-high-pressure liquid chromatography-tandem mass spectrometry for fentanyl and norfentanyl after solid-phase extraction. In one of the three samples tested, fentanyl and norfentanyl were detected at concentrations of 28 and 78 ng/mL, respectively. Based on the infusion rate, the theoretical oral fluid fentanyl concentration at steady state was calculated to be 33 ng/mL., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Objectives: Urine propylene glycol (PG) and vegetable glycerin (VG) were evaluated as potential markers for discriminating ECIG users from non-users and verifying ECIG abstinence., Methods: Urine samples from 51 ECIG users (collected pre/post 12-hours ECIG abstinence), and 50 controls (who do not use nicotine/tobacco) were analyzed for urine cotinine, PG, and VG concentration., Results: Of 42 ECIG users with pre-abstinence urine cotinine indicating nicotine use, mean (SD) urine cotinine concentration was 1053.7 ng/ml (874.5) and for controls was 1.93 ng/ml (0.4); after abstinence, ECIG users' mean cotinine decreased to 615.4 ng/ml (753.0). For ECIG users, mean urine PG pre-abstinence was 25.6 mcg/ml (20.0) and was 9.8 mcg/ml (13.5) for controls; after abstinence, ECIG users' mean urine PG decreased to 9.7 mcg/ml (15.0; ps < .05). For ECIG users, mean urine VG pre-abstinence was 7.5 mcg/ml (7.1) and was 13.2 mcg/ml (25.0) for controls; after abstinence, ECIG users' mean VG decreased to 5.0 mcg/ml (4.4; ps < .05)., Conclusions: ECIG users' mean urine PG was greater than controls and decreased after 12-hours ECIG abstinence suggesting urine PG may be useful for discriminating ECIG users from non-users and verifying short-term abstinence., Competing Interests: Conflict of Interest Statement Dr. Eissenberg is a paid consultant in litigation against the tobacco industry and electronic cigarette industry and is named on a patent application for a device that measures the puffing behavior of electronic cigarette users. All other authors have no disclosures.

Use of marijuana and cannabinoids has been on the rise in recent years, including in childbearing women. This has resulted in cannabinoids being more frequently identified in breast milk as a result of its high lipid content and cannabinoids having a high lipophilicity, thereby exposing the breastfeeding infant to cannabinoids and other marijuana constituents. Presented is a method for the analysis of Δ9-tetrahydrocannabinol (THC), cannabinol (CBN) and cannabidiol (CBD) in breast milk. THC, CBN, CBD and their isotopically labeled standards were extracted from breast milk using a modified QuEChERS method and analyzed using ultra-performance liquid chromatography and tandem mass spectrometry. As a result of the high lipid content of breast milk, saponification of the lipids was necessary to improve overall extraction efficiency. The process efficiency percentage for THC, CBD and CBN are 55%, 80% and 25%, respectively. The recovery percentage for THC, CBD and CBN are 95%, 118% and 85%, respectively. The matrix effect percentage for THC, CBD and CBN are 53%, 66% and 26%, respectively. Linearity was assessed from 1 to 100 ng/mL for THC, CBN and CBD and had r2 > 0.996. Validation controls were prepared at 1, 3, 20, 80 and 300 ng/mL (dilution control), and the bias was determined to be less than ±20% with %CVs <15% for all controls. Due to the limited access of genuine breast milk for routinely preparing matrix matched calibration and control materials, Enfamil® Premium™ Newborn Infant Formula (0-3 months) was evaluated as a breast milk substitute. No significant differences were observed for THC, CBN and CBD using either breast milk or formula as the matrix; thus, it was determined to be an acceptable breast milk matrix substitute. The modified QuEChERS method was determined to be a robust, reliable method for the determination of THC, CBN and CBD in breast milk., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Presented is an ultra-high-pressure liquid chromatographic tandem mass spectrometry (UPLC-MS/MS) method developed for the detection of propylene glycol, glycerol, ethylene glycol and diethylene glycol using isotopically labeled standards in urine as part of ongoing studies to evaluate whether urinary propylene glycol and/or vegetable glycerin concentration are indicators of recent use. Propylene glycol and vegetable glycerol are found in many products that are consumed and used including electronic cigarettes (e-cigarettes). E-cigarettes are battery-powered devices used as an alternative to traditional cigarettes. The liquid formulations aerosolized in these devices largely consist of propylene glycol and/or vegetable glycerol. Published reports regarding the ratio of propylene glycol to glycerol content in these formulations ranged from 50:50 to 100 percent of either. For the analysis of urine specimens from both users and non-users of e-cigarettes, calibrators, controls and specimens were derivatized using benzoyl chloride prior to analysis. They were analyzed using a Waters AcQuity Xevo TQ-S Micro UPLC-MS/MS. Chromatographic separation was performed on an AcQuity UPLC BEH C18 1.7 um, 2.1 × 50 mm, column using a 20 mM ammonium formate in water and 20 mM ammonium formate in methanol as the mobile phase. The method was validated using SWGTOX guidelines for linearity, precision and accuracy, stability, carryover and limit of detection. The linear range was determined using a seven-point calibration curve ranging between 0.5 and 100 mcg/mL. The bias for all validation controls was determined to be ±20% of the expected concentrations with CVs of <15%. A total of 124 urine specimens analyzed collected with 50 specimens collected from self-reported non-smokers (cigarettes/e-cigarettes) confirmed cotinine free using the DRI® Cotinine Assay (Thermo Scientific, Waltham, MA) and 74 specimens collected before and after 12 hours self-reported e-cigarettes abstinence e-cigarette users. Propylene glycol and glycerol were determined to have concentration ranges of "none detected" to 1470 and "none detected" to 2950 mcg/mL, respectively., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Opioid usage in the USA has increased over the past decade, with prescriptions increasing from 76 million in 1991 to 207 million in 2013. New regulations have curbed the number of prescriptions, leading to an increase in heroin use. Heroin-related overdoses have quadrupled between 2000 and 2015. The traditional urinary biomarkers for indicating heroin use are a combination of morphine and 6-acetyl morphine (6-AM). Morphine is detectable in urine for several days. 6-AM is detected in urine for 2-8 hours. Papaverine has been proposed as an alternative heroin biomarker. It has been reported to have a 1-2 day detection window. Papaverine metabolites have been reported to have up to a 3-day detection window. Presented is a method for the detection of papaverine and its metabolites, 6-desmethyl papaverine (6-DMP) and 4', 6-didesmethyl papaverine (4,6-DDMP), in urine using a modified Waters® MCX™ microelution method. An ultra-performance liquid chromatography and tandem mass spectrometry (UPLC-MS-MS), with a Waters' BEH C18 column, and 20 mM ammonium formate water: 20 mM ammonium formate methanol mobile phase was employed. Calibration curves were linear from 0.1 to 50 ng/mL. No interferences were observed from the analysis of multicomponent therapeutic drug or drugs of abuse control materials; intra- and inter-run precision tests were acceptable. A total of 428 genuine urine specimens where heroin use was suspected were analyzed. These included 101 6-AM and 179 morphine only positive samples as well as 6 morphine-negative samples where papaverine and/or metabolites were detected. The determined concentrations in these samples for papaverine, 6-DMP and 4,6-DDMP ranged from 0.10 to 994, 0.10 to 462 and 0.12 to 218 ng/mL, respectively. The method was rugged and robust for the analysis of papaverine and metabolites, 6-DMP and 4,6-DDMP. The use papaverine and metabolites, 6-DMP and 4,6-DDMP has the potential to increase the detection window of heroin use., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Carbenoxolone is a derivative of glycyrrhetinic acid found in the root of Glycyrrhiza glabra, colloquially known as licorice. It has been used as a treatment for peptic and oral ulcers. In recent years, carbenoxolone has been utilized in basic research for its ability to block gap junctional communication. Better understanding the distribution of carbenoxolone after systemic administration can lead to a better understanding of its potential sites of action. Presented is an ultra high-performance liquid chromatography tandem mass spectrometer (UHPLC-MS/MS) method for the identification and quantification of carbenoxolone in mouse blood and brain tissue. Twenty mice were injected intraperitoneally with 25 mg/kg carbenoxolone and brain tissue and blood were collected for analysis. Blood concentrations (mean ± SD) at 15, 30, 60 and 120 min were determined to be (n = 5) 5394 ± 778, 2636 ± 836, 1564 ± 541 and 846 ± 252 ng/mL, respectively. Brain concentrations (mean ± SD) at 15, 30, 60 and 120 mins were determined to be (n = 5) 171 ± 62, 102 ± 35, 55 ± 10 and 27 ± 9 ng/g, respectively. The analysis of these specimens at the four different time points resulted in blood and brain half-lives in mice of ~43 and 41 min, respectively. The UHPLC-MS/MS method was determined to be sensitive and robust for quantification of carbenoxolone., (© 2018 John Wiley & Sons, Ltd.)

Personal battery-powered vaporizers or electronic cigarettes were developed as an alternative to traditional cigarettes. The modern electronic cigarettes were patented in 2004 by Hon Lik in China. In May 2016, the US Food and Drug Administration (FDA) imposed regulatory statutes on e-cigarettes and their liquid formulations (e-liquids); prior to that, they were unregulated. E-liquids are typically composed of propylene glycol and/or glycerin, flavouring component(s), and active ingredient(s), such as nicotine. Fifty-six commercially available e-liquids, purchased from various sources, contained a variety of flavours and active ingredients. A headspace gas chromatography with flame ionization detector (HS-GC-FID) method was used to analyze these e-liquids for volatiles content. Only one of the e-liquids listed ethanol as a component. The chromatographic separation of volatiles was performed on a Restek BAC-1 column. A linear calibration was generated for ethanol with limits of detection and quantification (LOD/LOQ) of 0.05 mg/mL. Ethanol concentrations in the 56 e-liquids ranged from none detected to 206 mg/mL. The ethanol determined in these products may have been used in flavourants or a solvent; the reason for inclusion cannot be fully ascertained. The implications of vaporizing ethanol as an e-liquid component are unknown. Copyright © 2017 John Wiley & Sons, Ltd., (Copyright © 2017 John Wiley & Sons, Ltd.)

The blue lotus flower (Nymphea caerulea) is an Egyptian water lily containing apomorphine and nuciferine. Apomorphine has been described as a psychoactive alkaloid and is a non-selective dopamine agonist primarily used to treat Parkinson's disease as it stimulates dopamine receptors and improves motor function. Nuciferine is an alkaloid associated with dopamine receptor blockade. Today, blue lotus flower is used as a sleep aid and anxiety reliever. The rebuildable dripping atomizer (RDA) is an electronic cigarette that allows direct application of an e-liquid onto the coil in the atomizer for aerosolization, compared to a typical electronic cigarette where the e-liquid is wicked from a storage vessel to the coil. Our laboratory received a dark-brown resin material from a concerned parent. The resin had been confiscated from an adolescent who had a reported history of marijuana use. The resin was later identified as blue lotus flower (N. caerulea). This resin, together with four commercially available blue lotus products, was analyzed for content. Apomorphine was detected in two samples, and nuciferine was detected in all five samples. The confiscated resin was determined to contain no apomorphine and 4300 ng/g of nuciferine. The nuciferine resin was shown to aerosolize using aRDA electric cigarette.

Tianeptine (7-[([3-chloro-6,11]-dihydro-6-methyldibenzo[c,f][1,2]thiazepin-11-yl) amino] heptanoic acid S, S dioxide) is a tricyclic compound prescribed as an antidepressant in European countries, but is not currently approved for use in the United States. There are few published case reports of tianeptine intoxication. Presented is the first case of acute toxicity associated with the intravenous use of tianeptine. A 36-year-old male intentionally injected tianeptine powder intravenously to "help him see into the future". He became unresponsive and a bystander called emergency medical services. Upon arrival to the Emergency Department, excessive constriction of the pupils, sedation, and a respiratory rate of 6 respirations per minute (rpm) were noted. Blood and urine were collected ~2 h post admission. The patient's serum ethanol concentration was 133 mg/dL. His toxicity was successfully reversed with two doses of naloxone 0.4 mg IV. He was started on a naloxone infusion at 0.2 mg/h and discharged 13 h after admittance awake, alert and oriented. The patient's urine sample screened negative for common drugs of abuse and total tricyclic antidepressants. A high performance liquid chromatography tandem mass spectrometry method was developed and validated to quantify tianeptine in urine. The calibration range was 1-100 ng/mL with linear regression correlation (r2) of 0.9996 or greater. The limit of quantitation was administratively set at 1 ng/mL. The bias of the assay was determined to be within ±20% of the target value for each quality control specimen. The intra-day and inter-day precision did not exceed 15% coefficient of variation for each quality control specimen. Matrix effects, absolute recovery, carryover and specificity were also evaluated. The patient's tianeptine urine concentration was determined to be 2 ng/mL., (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Purpose: Professional organizations provide no guidelines regarding assessment and management of opioid abuse risk in cancer. Universal precautions (UP) developed for non-cancer pain, include assessments for aberrant behavior, screening questionnaires, and urine drug screens (UDS). The role of UDS for identifying opioid abuse risk in cancer is uncertain. Our aim is to characterize inappropriate UDS, and identify a potential role for UDS in therapeutic decision-making., Methods: An observational retrospective chart review of 232 consecutive supportive care clinic patients were seen during the study. Twenty-eight of the two hundred thirty-two did not meet inclusion criteria. One hundred fifty of the two hundred four had active cancer, while 54 had no evidence of active disease. Clinicians ordered UDS based on their clinical judgment of patients' substance misuse risk. Edmonton symptom assessment scores, history of substance abuse, alcohol use, tobacco use, aberrant behavior, and morphine equivalent daily dose (MEDD) were obtained., Results: Pain scores and MEDD were higher (p = 0.021; p < 0.001) in the UDS group vs non-UDS. Forty percent of the patients (n = 82/204) had at least one UDS and 70% (60/82) had an inappropriate result. Thirty-nine percent (32) were inappropriately negative, showing no prescribed opioids. Forty-nine of the eighty-two were positive for non-prescribed opioids, benzodiazepine, or illicit substance. Eleven of the forty-nine had only cannabis metabolites in their urine. There were no significant differences between appropriate and inappropriate UDS groups regarding pain scores, MEDD or referral to psychology, psychiatry, or substance abuse specialists., Conclusions: UDS on the 82 oncology patients at high risk for substance misuse were frequently positive (46%) for non-prescribed opioids, benzodiazepines or potent illicit drugs such as heroin or cocaine, and 39% had inappropriately negative UDS, raising concerns for diversion.

The legalization of marijuana in the USA for both medicinal and recreational use has increased in the past few years. Currently, 24 states have legalized marijuana for medicinal use. The US Drug Enforcement Administration has classified marijuana as a Schedule I substance. The US Food and Drug Administration does not regulate formulations or packages of marijuana that are currently marketed in states that have legalized marijuana. Marijuana edibles or "medibles" are typically packages of candies and baked goods consumed for medicinal as well as recreational marijuana use. They contain major psychoactive drug in marijuana, delta-9-tetrahydrocannabinol (THC) and/or cannabidiol (CBD), which has reputed medical properties. Presented is a method for the preparation and application of THC and CBD containing brownies used as quality control (QC) material for the analysis of marijuana or cannabinoid baked medibles. The performance parameters of the assay including possible matrix effects and cannabinoid stability in the brownie QC over time are presented. It was determined that the process used to prepare and bake the brownie control material did not degrade the THC or CBD. The brownie matrix was found not to interfere with the analysis of a THC or a CBD. Ten commercially available brownie matrixes were evaluated for potential interferences; none of them were found to interfere with the analysis of THC or CBD. The laboratory baked medible QC material was found to be stable at room temperature for at least 3 months., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

MDMB-FUBINACA (aka MDMB(N)-Bz-F), chemical name Methyl (S)-2-(1-(4-fluorobenzyl)-1H-indazole-3-carboxamido)-3,3-dimethylbutanoate, a designer drug or a new psychoactive substance (NPS), was identified in three commercially available e-liquids formulated for electronic cigarette use. The e-liquids were evaluated using direct analysis in real time ion source attached to a time of flight mass spectrometer (DART-MS) and gas chromatograph mass spectrometer (GC-MS) to identify active ingredients/drugs, flavorants, and other possible constituents. The e-liquids were also evaluated for alcohol content by headspace gas chromatography with flame ionization detector (HS-GC-FID). The aerosol produced from the e-liquids by use of an e-cigarette was analyzed by solid phase micro-extraction gas chromatography mass spectrometry (SPME-GC-MS) to ensure delivery of the active ingredient/drug. Propylene glycol, vegetable glycerin, MDMB-FUBINACA, alcohol content and a flavor profile were determined for each of the e-liquids. MDMB-FUBINACA was determined to be the major active ingredient in all three e-liquids and was successfully detected by SPME-GC-MS in the aerosol generated by a KangerTech Aerotank clearomizer/electronic cigarette., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Ethanol is the most widely used and abused drug. While blood is the preferred specimen for analysis, tissue specimens such as brain serve as alternative specimens for alcohol analysis in post-mortem cases where blood is unavailable or contaminated. A method was developed using headspace gas chromatography with flame ionization detection (HS-GC-FID) for the detection and quantification of ethanol, acetone, isopropanol, methanol and n-propanol in brain tissue specimens. Unfixed volatile-free brain tissue specimens were obtained from the Department of Pathology at Virginia Commonwealth University. Calibrators and controls were prepared from 4-fold diluted homogenates of these brain tissue specimens, and were analyzed using t-butanol as the internal standard. The chromatographic separation was performed with a Restek BAC2 column. A linear calibration was generated for all analytes (mean r 2  > 0.9992) with the limits of detection and quantification of 100-110 mg/kg. Matrix effect from the brain tissue was determined by comparing the slopes of matrix prepared calibration curves with those of aqueous calibration curves; no significant differences were observed for ethanol, acetone, isopropanol, methanol and n-propanol. The bias and the CVs for all volatile controls were ≤10%. The method was also evaluated for carryover, selectivity, interferences, bench-top stability and freeze-thaw stability. The HS-GC-FID method was determined to be reliable and robust for the analysis of ethanol, acetone, isopropanol, methanol and n-propanol concentrations in brain tissue, effectively expanding the specimen options for post-mortem alcohol analysis., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

In healthcare settings drug diversion and impairment of physicians are major concerns requiring a rapid and efficient method for surveillance and detection. A Direct Analysis in Real Time ion source coupled to a JEOL AccuTOF TM time-of-flight mass spectrometer (DART-MS) method was developed to screen parenteral pharmaceutical formulations for potential drug diversion. Parenteral pharmaceutical formulations are also known as injectable formulations and are used with intravenous, subcutaneous, intramuscular and intra-articular administration. A library was created using the mass spectra data collected by a DART-MS operated in switching mode at 20, 60 and 90 V settings. This library contained 17 commonly encountered drugs in parenteral pharmaceutical formulations that included the surgical analgesic: fentanyl, hydromorphone and morphine; anesthetic: baclofen, bupivacaine, ketamine, midazolam, ropivacaine and succinylcholine; and a mixture of other drug classes: caffeine, clonidine, dexamethasone, ephedrine, heparin, methadone, oxytocin and phenylephrine. Randomly selected 200 de-identified parenteral pharmaceutical formulations containing one or more drugs were submitted for analysis to the FIRM Toxicology Laboratory at Virginia Commonwealth University Health and were screened using the DART-MS. The drug contents of the de-identified formulations were previously confirmed by a published high performance liquid chromatography (HPLC) method. The drugs in the formulations were rapidly and successfully identified using the generated library. The DART-MS and HPLC results were in complete agreement for all 200 parenteral pharmaceutical formulations., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

A new Hydrocodone Enzyme Immunoassay (HEIA; Lin-Zhi International, Inc.) was evaluated for the detection of hydrocodone and its main metabolite, hydromorphone. All specimens were tested with two different cutoff calibrators, 100 and 300 ng/mL, on an ARCHITECT Plus c4000 Clinical Chemistry Analyzer. Controls containing -25% (negative control) and +25% (positive control) of the cutoff calibrators and a drug-free control were analyzed with each batch. All 1,025 urine specimens were previously analyzed by ultra-performance liquid chromatography-mass spectrometry/mass spectrometry (UPLC-MS-MS) for opiates. Approximately, 33% (337/1,019) of the specimens yielded positive results by the HEIA assay at a cutoff concentration of 100 ng/mL and 19% (190/1,025) yielded positive results at the 300 ng/mL cutoff concentration. Of these presumptive positive specimens, UPLC-MS-MS confirmed the presence of hydrocodone and/or hydromorphone >100 ng/mL in 241 specimens and >300 ng/mL in 162 specimens, for each respective cutoff. With the 100 ng/mL cutoff, the HEIA demonstrated a sensitivity of 0.959, a specificity of 0.846 and an overall agreement with the UPLC-MS-MS of 87%. At 300 ng/mL cutoff, the HEIA demonstrated a sensitivity of 0.880, a specificity of 0.966 and an overall agreement of UPLC-MS-MS results of 95%. The Lin-Zhi HEIA 100 ng/mL cutoff assay demonstrated sensitivity for the detection of hydrocodone and hydromorphone in urine. The 300 ng/mL cutoff was less sensitive, but more selective, and should be part of an initial immunoassay screen, particularly in pain management compliance testing., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Background: Adulteration of drugs of abuse may be done to increase profits. Some adulterants are relatively innocuous and others result in significant toxicity. Clenbuterol is a β2-adrenergic agonist with veterinary uses that has not been approved by the U.S. Food and Drug Administration for human use. It is an infrequently reported heroin adulterant. We describe a cluster of hospitalized patients with laboratory-confirmed clenbuterol exposure resulting in serious clinical effects., Case Series: Ten patients presented with unexpected symptoms shortly after heroin use. Seven evaluated by our medical toxicology service are summarized. Presenting symptoms included chest pain, dyspnea, palpitations, and nausea/vomiting. All patients were male, with a median age of 40 years (interquartile range [IQR] 38-46 years). Initial vital signs included a heart rate of 120 beats/min (IQR 91-137 beats/min), a respiratory rate of 20 breaths/min (IQR 18-22 breaths/min), a temperature of 36.8°C (IQR 36.7-37.0°C), a systolic blood pressure of 107 mm Hg (IQR 91-131 mm Hg), and a diastolic blood pressure of 49 mm Hg (IQR 40-70 mm Hg). Serum potassium nadir was 2.5 mEq/L (IQR 2.2-2.6 mEq/L), initial glucose was 179 mg/dL (IQR 125-231 mg/dL), initial lactate was 9.4 mmol/L (IQR 4.7-10.5 mmol/L), and peak creatine phosphokinase was 953 units/L (IQR 367-10,363 units/L). The median peak troponin level in six patients was 0.7 ng/mL (IQR 0.3-2.4 ng/mL). Three patients underwent cardiac catheterization and none had significant coronary artery disease. Clenbuterol was detected in all patients after comprehensive testing. All patients survived with supportive care. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Atypical presentations of illicit drug intoxication may raise concern for drug adulteration. In the case of heroin use, the presence of adrenergic symptoms or chest pain with hypokalemia, lactic acidosis, and hyperglycemia suggests adulteration with a β-agonist, such as clenbuterol, and patients presenting with these symptoms often require hospitalization., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Since 24 states and the District of Columbia have legalized marijuana in some form, suppliers of legal marijuana have developed Cannabis sativa products for use in electronic cigarettes (e-cigarettes). Personal battery powered vaporizers, or e-cigarettes, were developed to deliver a nicotine vapor such that smokers could simulate smoking tobacco without the inherent pathology of inhaled tobacco smoke. The liquid formulations used in these devices are comprised of an active ingredient such as nicotine mixed with vegetable glycerin (VG) and/or propylene glycol (PG) and flavorings. A significant active ingredient of C. sativa, cannabidiol (CBD), has been purported to have anti-convulsant, anti-nociceptive, and anti-psychotic properties. These properties have potential medical therapies such as intervention of addictive behaviors, treatments for epilepsy, management of pain for cancer patients, and treatments for schizophrenia. However, CBD extracted from C. sativa remains a DEA Schedule I drug since it has not been approved by the FDA for medical purposes. Two commercially available e-cigarette liquid formulations reported to contain 3.3 mg/mL of CBD as the active ingredient were evaluated. These products are not regulated by the FDA in manufacturing or in labeling of the products and were found to contain 6.5 and 7.6 mg/mL of CBD in VG and PG with a variety of flavoring agents. Presently, while labeled as to content, the quality control of manufacturers and the relative safety of these products is uncertain.

Personal battery-powered vaporizers or electronic cigarettes were developed to deliver a nicotine vapor such that smokers could simulate smoking tobacco without the inherent pathology of inhaled tobacco smoke. Electronic cigarettes and their e-cigarette liquid formulations are virtually unregulated. These formulations are typically composed of propylene glycol and/or glycerin, flavoring components and an active drug, such as nicotine. Twenty-seven e-cigarette liquid formulations that contain nicotine between 6 and 22 mg/L were acquired within the USA and analyzed by various methods to determine their contents. They were screened by Direct Analysis in Real Time™ Mass Spectrometry (DART-MS). Nicotine was confirmed and quantitated by high-performance liquid chromatography-tandem mass spectrometry, and the glycol composition was confirmed and quantitated by gas chromatography-mass spectrometry. The DART-MS screening method was able to consistently identify the exact mass peaks resulting from the protonated molecular ion of nicotine, glycol and a number of flavor additives within 5 mmu. Nicotine concentrations were determined to range from 45 to 131% of the stated label concentration, with 18 of the 27 have >10% variance. Glycol composition was generally accurate to the product description, with only one exception where the propylene glycol to glycerin percentage ratio was stated as 50:50 and the determined concentration of propylene glycol to glycerin was 81:19 (% v/v). No unlabeled glycols were detected in these formulations., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Ivabradine is a newly approved medication which reduces the heart rate by antagonizing the If channel. We report a case of intentional overdose on ivabradine. A 26-year-old female presented after taking 250 mg ivabradine. On arrival, her vital signs and neurologic exam were unremarkable. Within 30 min, her heart rate decreased to 31 bpm, but she remained normotensive with no change in mentation. Her bradycardia resolved after treatment with atropine. She experienced two further bradycardic episodes responsive to atropine; the second episode was associated with hypotension, responsive to a fluid bolus. For the remainder of her hospitalization, she remained hemodynamically stable without further interventions. She was dispositioned to the psychiatry service approximately 36 h post-ingestion with a heart rate of 67 bpm. Laboratory analysis confirmed a serum ivabradine concentration of 525 ng/mL, greater than 50 times the mean level in therapeutic trials. Proposed treatments for ivabradine include activated charcoal, atropine, isoproterenol, and intravenous pacing. Further study is needed to identify ideal treatment modalities.

In recent years, a large number of designer drugs sold as "Bath Salts" have appeared on the market. In July of 2011, Raving Dragon Novelty Bath Salts was obtained over the Internet. This product became unavailable in October of that year coinciding with the DEA issuing a temporarily schedule of mephedrone, methylone, and MDPV. Four months later in February of 2012, a new product was released from the same company under the new name Raving Dragon Voodoo Dust. The contents of both products were identified using spectroscopy methods: nuclear magnetic resonance, infrared, UV-visible, tandem mass spectrometry, and high-resolution time-of-flight mass spectrometry. It was determined that Raving Dragon Novelty Bath Salts contained methylone. The replacement product Raving Dragon Voodoo Dust contained the unscheduled drug pentedrone. The Raving Dragon brand of products illustrates the rapid change of ingredients in these products to circumvent laws restricting availability, distribution, and use., (© 2014 American Academy of Forensic Sciences.)

We present a case of 4-bromo-2,5-dimethoxy-N-[(2-methoxyphenyl)methyl]-benzeneethanamine (25B-NBOMe), an N-benzyl phenethylamines derivative, intoxication and a high performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) method for detection and quantification of 25B-NBOMe. A 19-year-old male was found unresponsive with generalized grand mal seizure activity. On the second day of hospitalization, a friend admitted that the patient used 'some unknown drug' called 25B. Serum and urine collected were sent to the Virginia Commonwealth University Medical Center Toxicology Laboratory for analysis. An HPLC-MS/MS method for the identification and quantification of 25B-NBOMe using 2-(2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine (25H-NBOMe) as the internal standard (ISTD) was developed. As this is a novel, single-case presentation, an assay validation was performed prior to testing to ensure the reliability of the analytical results. The serum and urine specimens were determined to contain 180 pg/ml and1900 pg/ml of 25B-NBOMe, respectively., (Copyright © 2013 John Wiley & Sons, Ltd.)

Background: We evaluated a new EDDP [2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine] enzyme immunoassay (EDDPI; Lin-Zhi International, Inc., Sunnyvale, CA) for the detection of this primary methadone urinary metabolite., Methods: All specimens were tested with two different cutoff calibrators at 150 and 300 ng/ml EDDP on an ADVIA 1200 Chemistry System auto-analyzer. Controls containing 0, -25% (negative control), and +25% (positive control) of the cutoff calibrators (Lin-Zhi) were analyzed with each batch. All urine specimens were then analyzed by high-pressure liquid chromatography/ mass spectrometry/mass spectrometry (HPLC-MS/MS) for EDDP., Results: Approximately, 42% (151) of the 362 specimens yielded positive results by the EDDP assay at 150 and/or 300 ng/ml cutoff values. Of these specimens, HPLC-MS/MS confirmed the presence of EDDP > 25 ng/ml in all 151 specimens. No specimen yielding negative EDDPI results contained EDDP by HPLC/MS/MS. At 150 ng/ml cutoff, the EDDPI demonstrated a sensitivity of 1.00, a specificity of 0.986, and an overall agreement of HPLC/MS/MS of >99%. At 300 ng/ml cutoff, the EDDPI demonstrated a sensitivity of 1.00, a specificity of 0.959, and an overall agreement of HPLC/MS/MS results of 97.5%., Conclusion: The Lin-Zhi EDDPI provides a precise, reliable method for the routine detection of methadone metabolite in urine specimens, particularly in pain management compliance testing., (© 2014 Wiley Periodicals, Inc.)

2CC-NBOMe {4-chloro-2,5-dimethoxyphenethyl-N-[(2-methoxyphenyl) methyl] ethanamine} and 25I-NBOMe {2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl) methyl] ethanamine} are of a class of N-benzyl phenethylamine derivatives whose synthesis was first reported in the scientific literature in 2011. Recent reports from 'personal drug experience websites' and in the popular press indicate these drugs are the latest in a series of designer 'Bath Salt' drugs of abuse. The presented high-performance liquid chromatography triple quadrupole mass spectrometry (HPLC/MS/MS) method was developed for the detection and quantification of 2CC-NBOMe and 25I-NBOMe in serum of intoxicated emergency department patients. The assay applies 2-​(2,​5-​dimethoxyphenyl)-​N-​(2-​methoxybenzyl) ethanamine (25H-NBOMe) as the internal standard. Samples were extracted using solid-phase extraction columns. The chromatographic separation was performed on a Luna 3 µ C8(2) 100 Å, 100 × 2.0 mm, column. Detection was accomplished by multiple-reaction monitoring via an electrospray ionization source operating in the positive ionization mode. The calibration curves were linear over the investigated concentration range, 30-2000 pg/mL, with a lower limit of detection of 10 pg/mL for both 2CC-NBOMe and 25I-NBOMe. The method proved suitable for serum clinical toxicology testing. Two severely intoxicated emergency department patients were determined to have serum concentrations of 250 and 2780 pg/mL of 25I-NBOMe using the presented method., (Copyright © 2013 John Wiley & Sons, Ltd.)

Background: Benzoylecgonine (BE) is the primary urinary metabolite of cocaine. Two enzyme immunoassays were evaluated for the detection of BEin urine with a 300 ng/ml cutoff: the DRI® Cocaine Metabolite Assay and Lin-Zhi International's (LZ) Cocaine Metabolite Enzyme Immunoassay., Methods: This study involved 1,398 urine specimens from criminal justice and pain management programs. Gas chromatography/mass spectrometry (GC/MS) data were obtained for presumptive positives, and for negative urine specimens yielding responses significantly above the negative control., Results: Approximately 46% (644) of the specimens yielded positive results by DRI, and 47% (664) were positive by LZ. One specimen screened positive with both assays but was found to have a nondetectable BE concentration by GC/MS, indicating one false positive for each assay. Twenty-one specimens yielding negative DRIresults contained BEabove 300 ng/ml, and 29 specimens yielded false negatives with the LZassay. Therefore, the overall agreement between both immunoassays and GC/MSresults was 98%. Assay sensitivity was 0.968 (DRI) and 0.958 (LZ); the selectivity for both assays was 0.999. Urine specimens containing cocaine, additional cocaine metabolites, and other drugs were also tested. No cross-reactivity was observed., Conclusion: Both the DRIand LZassays provide a precise, reliable method for the routine detection of BEin urine., (© 2012 Wiley Periodicals, Inc.)

Background: Guanfacine, an α2-adrenergic α2A)agonist long indicated to treat hypertension, is now being used to treat attention deficit-hyperactivity disorder (ADHD) in adolescents. This new therapeutic use may require urine testing to document compliance or abuse. A simple rapid high pressure liquid chromatography-mass spectrometry (HPLC-MS) method to detect and quantify guanfacine in urine following therapeutic administration is presented., Methods: Guanfacine and protriptyline internal standard were extracted from alkalinized urine with ethyl acetate. The organic layer was evaporated, reconstituted with mobile phase and analyzed on a YMC Basic S-5 micron, 2.0 × 150 mm HPLC column connected to an MS detector operated in positive electrospray ionization mode with selected ion resonance. Elution times were <5 min., Results: The analytical measurement range for guanfacine was 20-2000 ng/mL. The limit of detection and quantitation were 5 ng/mL and 20 ng/mL, respectively. Precision as %CV was <15% at 40, 100 and 500 ng/mL (n=6). Percentage recovery using the same concentrations was >89%. Interference with drugs and biological constituents was assessed; no interferences were noted. Analysis of 100 random post-diagnostic urine specimens yielded 11 guanfacine positive results with concentrations ranging from 11 to 6390 ng/mL., Conclusions: This HPLC-MS method provides a simple and rapid method for the routine detection and quantitation of guanfacine in urine specimens.

Cannabichromene (CBC) is a phytocannabinoid, the second most abundant cannabinoid quantitatively in marijuana. CBC has been shown to produce antinociception and anti-inflammatory effects in rodents. This method is validated for the measurement of THC and CBC simultaneously after extraction from mouse blood or brain. Whole brain harvested from mice was homogenized 2:1 (v/w) with normal saline. Fifty nanograms of THC-d₃ was added to 0.5 mL of heparinized mouse blood, brain homogenate, and THC and CBC fortified blood or brain calibrators, then equilibrated overnight at 5 °C. Two milliliters of "ice cold" acetonitrile was added drop-wise while the sample was vortex mixed, and then the sample was centrifuged and stored overnight at -30 °C. The cannabinoids were extracted from the acetonitrile layer with 2 mL of 0.2 N NaOH and 4 mL of hexane/ethyl acetate (9:1). The solvent was isolated and evaporated to dryness. Trimethylsilyl derivatives were prepared and then analyzed by gas chromatography-mass spectrometry. Linearity in blood and brain of THC and CBC was 2-10,000 ng/mL (ng/g). THC and CBC recovery ranged from 56 to 78% in blood and brain. Precision was demonstrated at 100 ng/mL and 1000 ng/mL with CVs < 15%. The validated method allows for blood and brain concentrations of cannabinoids to be quantificated and correlated with pharmacological effects produced in mice.

We present an evaluation of a new phencyclidine (PCP) enzyme immunoassay (PCPI) for detection in urine. The PCPI was evaluated by testing 523 urine specimens. Controls containing 0 ng/mL of PCP and -25% (negative control) and +25% (positive control) of the 25 ng/mL cutoff calibrator were analyzed with each batch. All urines were analyzed by high-performance liquid chromatography- tandem mass spectrometry (HPLC-MS-MS) for PCP. Of the 523 specimens tested, 218 yielded positive results by the PCP assay. HPLC-MS-MS confirmed the presence of at least 25 ng/mL in 214 specimens, indicating four false-positive results containing 11, 13, 16, and 20 ng/mL PCP. Three specimens yielded a negative result; however, PCP concentrations were within 20% of the 25 ng/mL cutoff value. The overall agreement of PCPI and HPLC-MS-MS results was 98.7%. The sensitivity of the PCPI was 0.982 and the specificity 0.987. Testing at 100 ng/mL of other drugs or their metabolites demonstrated no cross-reactivity. The within-run precision was CV = 2% (n = 12); the between-run precision was CV = < 6% (n = 4). The assay was found linear from -50% to 150% (12.5-37.5 ng/mL) of the cutoff concentration. The Lin-Zhi PCPI provides a precise, reliable method for the routine detection of phencyclidine in urine specimens.

A simple high-performance liquid chromatographic (HPLC) method that allows for the rapid identification and quantification of analgesic and anesthetic solutions typically used in surgical procedures or patient controlled analgesia is presented. The separation of bupivacaine, clonidine, fentanyl, hydromorphone, midazolam, and morphine is complete in less than 20 min. The method allows test solutions to be either directly injected or diluted prior to injection into the HPLC system. The method is useful from the standpoint that pharmaceutical preparations are usually submitted with the known drug of interest and expected concentration. The method is also useful for initial screening of solutions submitted that are either unknown or of questionable identity. The method has been successfully applied as part of hospital-based quality control and quality assurance programs to detect not only errors in the preparation of solutions of scheduled drugs, but also to uncover illegal diversion of drugs of abuse by medical personnel.

Background and Objectives: Leukapheresis of non-mobilized healthy donors is performed to harvest monocytes and lymphocyte subpopulations for use in various therapeutic regimens. In this methodological study, we compared two different leukapheresis programs, using equivalent volumes of processed blood over similar processing periods, to determine the influence of the procedures on the donor peripheral blood count and to establish the procedure that yields the highest quality product., Materials and Methods: The target variables obtained in 41 healthy blood donors who underwent short-term leukapheresis (80-105 min) were retrospectively compared. Twenty-one volunteers were processed on a COBE Spectra machine at the MNC setting and 20 volunteers were processed at the AutoPBSC setting. Data were collected on pre- and postleukapheresis samples and on the product., Results: AutoPBSC and MNC procedures resulted in a decrease of haemoglobin (5-7%), platelets (17-20%), monocytes (22%) and lymphocytes (23-27%), but not of granulocytes in peripheral blood. Both procedures produced nearly identical leucocyte and lymphocyte yields. AutoPBSC products contained a greater number of granulocytes, monocytes and red cells, but fewer platelets. The preleukapheresis values correlated with the yields for monocytes, T-helper and T-suppressor cells, B-lymphocytes and natural killer cells, but not for granulocytes or platelets., Conclusions: Leukapheresis is a safe and efficient procedure for collecting large numbers of peripheral blood monocytes and different lymphocyte populations from non-mobilized donors. The two programs yield comparable leucocyte harvests. Based on our results, yields can be predicted from the peripheral cell counts.

Objectives: In the past, some moonshine products contained potentially toxic contaminants. Although moonshine production continues in the United States, no studies have analyzed the content of moonshine since the early 1960s. We hypothesize that moonshine continues to contain potentially toxic concentrations of contaminants., Methods: Forty-eight samples of illicitly distilled moonshine were obtained from law enforcement agencies. An independent laboratory, blinded to both the moonshine source and a control sample of ethanol, conducted the analysis. Lead content was determined using atomic absorption spectrophotometry with a graphite tube atomizer. Alcohol content, including ethanol, acetone, isopropanol, methanol, and ethylene glycol, was determined using gas liquid chromatography with flame ionization detection., Results: Ethanol content ranged from 10.5% to 66.0% with a mean value of 41.2%. Lead was found in measurable quantities in 43 of 48 samples with values ranging from 5 to 599 parts per billion (ppb) with a mean value of 80.7 ppb. A total of 29 of 48 (60%) of samples contained lead concentrations above or equal to the EPA water guideline of 15 ppb. Methanol was found in only one sample at a concentration of 0.11%. No samples contained detectable concentrations of acetone, isopropanol, or ethylene glycol., Conclusions: Many moonshine samples contain detectable concentrations of lead. Extrapolations based on the described moonshine lead content suggest that chronic consumers of moonshine may develop elevated lead concentrations. Physicians should consider lead toxicity in the differential diagnosis when evaluating patients consuming moonshine.

Thrombophilic disorders and hypofibrinolysis were demonstrated to be risk factors in a majority of women with recurrent pregnancy loss (RPL) and infertility. We investigated the association of FV G1691A mutation, F II G20210A gene polymorphism (PM), 4G/5G PAI-1 and Alu I/D tPA PM in 32 women with infertility and 49 women with at least 2 unexplained early abortions. FV Leiden mutation was significantly more common in women with RPL (10%, p = 0.02) and infertility (19%, p = 0.0005) compared with controls (2%). PAI-1 4G PM and t-PA Alu I PM, alone or in combination, were not associated with RPL or infertility. 9/49 women with RPL showed coagulation disorders with heterozygous FV Leiden mutation (5), FXII (1), protein C (1) or protein S (2) deficiency. However, due to the small number of patients studied, no definite conclusion can be drawn., (Copyright 2003 S. Karger AG, Basel)

A rapid gas chromatographic method for the routine determination in serum of the new anticonvulsant drug topiramate (Topamax) (TOP) is described. The method involves extracting 0.50 mL of sample, previously adjusted to pH 9.5 with saturated borate buffer with ethyl acetate. One-microliter aliquots of the extract were injected into a 10-m x 0.53-mm i.d. x 0.5-microm 100% methyl silicone megabore capillary column connected to a nitrogen-phosphorus detector. The column temperature was initially at 170 degrees C for 0.1 min, then programmed at 10 degrees C/min to 240 degrees C, then 20 degrees C/min to 280 degrees C for 0.5 min. Under these conditions of the assay, the retention times of TOP and mepivicaine, internal standard, were 4.0 and 3.4 min, respectively. Quantitative determinations were performed with peak-height ratios of TOP to the internal standard. Calibration curves were linear from 2.5 to 150 mg/L TOP. The assay had a limit of quantitation of 2.5 mg/L. The overall within-run precision of the method yielded coefficients of variation (CV) of 3.9% at 10 mg/L (n = 10) and 3.1% at 100 mg/L (n = 10). The overall between-run precision calculated by three determinations on a single day for a week yielded CVs of 7.3% at 23 mg/L (n = 12) and 7.8% at 85 mg/L (n = 12). Common anticonvulsant and basic/neutral extractable drugs were found not to interfere with the assay. At present, no correlation has been demonstrated between trough plasma TOP concentrations and clinical efficacy. However, TOP values observed in our laboratory in serums from patients receiving adjunctive treatment for seizure disorders ranged from 2.5 to 35 mg/L.