Residual contamination in antineoplastic drug packaging.

INTRODUCTION: The handling of antineoplastic drugs should follow strict supervision and safety rules to minimize the occupational exposure risks to professionals involved. The external surface contamination of drug vials is recognized as a health risk. So, our goal was to determine if there is residual contamination on the vials and containers surface of the antineoplastic drugs doxorubicin (DOX) and cyclophosphamide (CP). METHODS: A cross-sectional study was conducted. Samples were collected using a uniform sampling procedure on the inner surfaces of the packages/boxes and the outer surfaces of the vials. The analyzes were executed by high-performance liquid chromatography/mass spectrometry (UHPLC-MS/MS). RESULTS: A total of 209 samples were analyzed, 66 of CP and 143 of DOX. CP levels were detected in nine samples (13.63%), three were below the lower limit of quantification (LLQ) and the other six had contamination levels ranging from 1.24 to 28.04 ng/filter. DOX levels were detected in 36 samples (25.17%), two were below the LLQ and the others had levels between 1.32 and 664.84 ng/filter. The majority of samples with residual contamination were in vials (80.0%), however, boxes also showed contamination. CONCLUSIONS: The results revealed the presence of residual contamination in the vials and packages of CP and DOX drugs. Although the residues found in each sample are small, special care should be taken in the handling and disposal of the antineoplastic drugs. The use of personal protective equipment is fundamental while handling the vials and packaging of cytotoxic drugs.

Simple extraction of toxicologically relevant psychotropic compounds and metabolites from whole blood using mini-QuEChERS followed by UPLC-MS/MS analysis.

The determination of psychotropic drugs and metabolites in blood is relevant in the context of both therapeutic drug monitoring and clinical and forensic toxicology. LC-MS/MS is the preferred method for these assays. However, LC-MS/MS is particularly susceptible to matrix ionization effects and appropriate sample preparation is required to minimize these effects. In this study, a simple, single-step, mini-QuEchERS extraction procedure, coupled to UPLC-MS/MS, was developed and validated for the determination of 15 toxicologically relevant compounds in whole blood, including psychoactive drugs and some metabolites. The assay was linear in the range of 25-1,000 ng ml-1 , fulfilling criteria for accuracy and precision. Extraction yields (71.9-87.7%) and matrix effects (-3.3 to +4.4%, with the exception of codeine, which had matrix effects of -35.36 to -28.14%) were acceptable for the majority of the evaluated compounds, using a single internal standard. The assay was applied to 238 clinical specimens from patients admitted to an emergency service, with 22 samples presenting quantifiable concentrations of 11 different compounds. The developed assay is a simple and efficient strategy for determination of target psychotropic drugs and metabolites in forensic and clinical toxicology.

A quick UPLC-MS/MS method for therapeutic drug monitoring of abiraterone and delta(4)-abiraterone in human plasma.

Abiraterone acetate efficacy against prostate cancer is dependent on the circulating levels of abiraterone and its active metabolites, which present significant pharmacokinetic variability among patients. Thus, therapeutic drug monitoring can be performed to improve treatment outcomes. To support such studies, there are only a limited number of bioanalytical methods in current literature. This work presents a fast method to quantify abiraterone and D4A in plasma in 4 min by UPLC-MS/MS. Bioanalytical method validation was performed according to the recommendations of the US Food and Drug Administration. The method was linear within the range of 1-400 ng/ml for abiraterone and 0.2-20 ng/ml for D4A (r2 > 0.99). Based on the analysis of quality control samples at the lower limit of quantification, low, medium and high concentrations, the method was precise (CVabiraterone ≤ 9.72%; CVD4A ≤ 14.64%) and accurate (CVabiraterone 95.51-107.59%; CVD4A 98.04-99.89%). Application of the method to the quantification of abiraterone and D4A in 10 clinical samples revealed important variability in the conversion ratio of abiraterone to D4A (CV 90.85%). Considering the current literature, this is the fastest method to quantify abiraterone and D4A in plasma, allowing for optimization of the analytical routine.

Determination of cannabinoids in plasma using salting-out-assisted liquid-liquid extraction followed by LC-MS/MS analysis.

The detection of the markers of Cannabis consumption in biological specimens is an important task for drug testing laboratories in varous contexts. A simple assay combining salting-out assisted liquid-liquid extraction sample preparation and LC-MS/MS analysis was applied to the measurement of Δ9 -tetrahydrocannabinol, 11-nor-9-carboxy-Δ9 -tetrahydrocannabinol (THC-COOH), 11-hydroxy-Δ9 -tetrahydrocannabinol, cannabinol and cannabidiol concentrations in 100 µl plasma specimens. The assay had linearity of 1-100 ng ml-1 for THC-COOH and 0.5-50 ng ml-1 for the other tested cannabinoids. Assay validation criteria were fulfilled. Extraction yields (88.7-97.3%) and internal-standard correct matrix effects (-9.6 to +5.4%) were acceptable. The assay was applied to 238 clinical specimens from trauma patients, with 19 samples presenting quantifiable concentrations of at least one of the target compounds. The developed assay is a simple and efficient strategy for simultaneous measurement of Δ9 -tetrahydrocannabinol, THC-COOH, 11-hydroxy-Δ9 -tetrahydrocannabinol, cannabinol and cannabidiol concentrations in plasma specimens.

Determination of Endogenous Concentrations of Uracil and Dihydrouracil in Dried Saliva Spots by LC-MS/MS: Method Development, Validation, and Clinical Application.

BACKGROUND: The aim of this study was to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for the measurement of uracil (U) and dihydrouracil (UH2) concentrations in dried saliva spots (DSSs), for the evaluation of dihydropyrimidine dehydrogenase (DPD) enzyme activity. RESULTS: Nine 18-mm diameter DSS discs were extracted with acetate:isopropyl alcohol (85:15, vol/vol) and analyzed by LC-MS/MS. The assay was linear in the range of 10-1000 ng·mL, with accuracy between 89% and 112% and precision between 5.7% and 13%. The metabolic ratio [UH2]/[U] was stable in DSS for up to 9 days at 45°C. Concentrations of U and UH2, as well as the metabolic ratio, were highly concordant between matrices. Using a metabolic ratio classification cutoff of 1.16 for the identification of slow DPD metabolizers, 98.7% concordance was achieved between SS and saliva. CONCLUSIONS: DSS samples could be a useful alternative for DPD activity screening, particularly in locations with limited access to highly equipped laboratories.

Evaluation of genotoxicity in workers exposed to low levels of formaldehyde in a furniture manufacturing facility.

Formaldehyde (FA) is a chemical widely used in the furniture industry and has been classified as a potential human carcinogen. The purpose of this study was to evaluate the occupational exposure of workers to FA at a furniture manufacturing facility and the relationship between environmental concentrations of FA, formic acid concentration in urine, and DNA damage. The sample consisted of 46 workers exposed to FA and a control group of 45 individuals with no history of occupational exposure. Environmental concentrations of FA were determined by high-performance liquid chromatography. Urinary formic acid concentrations were determined by gas chromatography with flame ionization detector. DNA damage was evaluated by the micronucleus (MN) test performed in exfoliated buccal cells and comet assay with venous blood. The 8-h time-weighted average of FA environmental concentration ranged from 0.03 ppm to 0.09 ppm at the plant, and the control group was exposed to a mean concentration of 0.012 ppm. Workers exposed to higher environmental FA concentrations had urinary formic acid concentrations significantly different from those of controls (31.85 mg L(-1) vs. 19.35 mg L(-), p ≤ 0.01 Mann-Whitney test). Significant differences were found between control and exposed groups for the following parameters: damage frequency and damage index in the comet assay, frequency of binucleated cells in the MN test, and formic acid concentration in urine. The frequency of micronuclei, nuclear buds, and karyorrhexis did not differ between groups. There was a positive correlation between environmental concentrations of FA and damage frequency (Spearman's rank correlation coefficient [r s] = 0.24), damage index (r s = 0.21), binucleated cells (r s = 0.34), and urinary formic acid concentration (r s = 0.63). The results indicate that, although workers in the furniture manufacturing facility were exposed to low environmental levels of FA, this agent contributes to the observed increase in cytogenetic damage. In addition, urinary formic acid concentrations correlated strongly with occupational exposure to FA.

Correlation analysis between cotinine hair concentrations from active smokers and nicotine intake and dependence.

OBJECTIVES: To determine cotinine levels in hair samples collected from smokers and estimate their relationship with nicotine intake and nicotine dependence class according to the Fagerström dependence questionnaire. DESIGN AND METHODS: Sixty-nine volunteers participated. They gave hair samples and answered questionnaires covering information related to smoking habits. Cotinine levels were determined by enzyme immunoassay after extraction from 20 mg of hair. Calibrators for the range 100 to 12,800 pg/mg were used. RESULTS: Mean cotinine concentration was 2070.9 pg/mg (range, 469.4-10,188.6 pg/mg). There were significant correlations between cotinine levels in hair and the questionnaire results (rs = 0.325, P = 0.018) and with the number of cigarettes smoked in a day (rs = 0.717, P < 0.001). Although the correlation between questionnaire results and the number of cigarettes smoked per day was significant (rs = 0.565, P < 0.01), it was weaker than the association between cotinine levels and number of cigarettes smoked. Cotinine levels were lower in the group that smoked from 1 to 5 cigarettes per day (1104.1 ng/mg) compared with the other groups. CONCLUSIONS: Cotinine levels were more highly correlated with nicotine exposure than with the Fagerström questionnaire scores. It was found that interference from exogenous sources of contamination, such as hair dyes, is a limitation to estimate nicotine intake from hair analysis.

Determination of meropenem in capillary plasma microsamples using LC-MS/MS.

Background: The measurement of meropenem plasma concentrations is employed for dosing regimen individualization. The aim of this study was to develop and validate a LC-MS/MS assay for quantification of meropenem in capillary plasma microsamples. Methods: Samples were prepared by protein precipitation with acetonitrile, followed by clean-up with dichloromethane. The method was validated and applied to 12 paired samples of venous and capillary plasma. Results: The method was linear in the range of 0.5-50 µg/ml. Matrix effects were minimal. Inter- and intra-assay were 3.8-7.9% and 2.7-5.5%, respectively, while accuracy was 91.7-100.6%. Concentrations in capillary and venous plasma were highly correlated. Conclusion: An assay for the quantification of meropenem in capillary plasma microsamples was fully validated, showing potential for clinical application.

[Box: see text].

GC-NPD analysis of topiramate in capillary dried plasma: assessing chemical dependency pharmacotherapy.

Aim: This study aimed to develop and validate a GC-NPD method for quantifying topiramate (TPM) in capillary dried plasma spots (DPS).Materials & methods: Extraction involved three 6 mm DPS with albumin 0.1%, alkaline liquid extraction with tert-Butyl methyl ether and TMAH methylation. The method was validated and applied to 15 paired samples of capillary DPS and venous plasma from chemical dependency patients.Results: The method was linear from 1 to 50 µg/ml (r >0.99), precise (CV% 3.62-8.29%) accurate (98.1-107.7%). TPM stability was confirmed in DPS stored at 4, 23 and 45°C for 21 days. DPS TPM measurements were highly correlated plasma concentrations (rs = 0.96), representing on average 102% of the venous plasma measurements.Conclusion: The method was fully validated, demonstrating potential for clinical application.

[Box: see text].

Determination of carbamazepine in fingerprints: a feasibility study to evaluate adherence in epilepsy patients.

Background: Fingerprint drug concentrations can be used as a noninvasive and convenient alternative to evaluate adherence to pharmacotherapy. Methods: Fingerprints were applied over glass slides, extracted and analyzed by ultra-high performance LC-MS/MS. The assay and drug adherence questionnaires were applied to 30 epilepsy patients. Results: The assay had linearity in the range 0.05-10 ng fingerprint-1, with precision of 2.16-7.9% and accuracy of 95.0-102.8%. Carbamazepine (CBZ) levels in fingerprints were stable at 45°C for 15 days. Concentrations in patient samples were 0.06-9.28 ng fingerprint-1. A significant difference (p = 0.003) was found between CBZ concentrations in fingerprints between patient groups divided as low and medium/high adherence. Conclusion: This method can potentially be applied to the identification of epilepsy patients with low adherence to CBZ pharmacotherapy.

[Box: see text].

Dried blood spot sampling for therapeutic drug monitoring: challenges and opportunities.

INTRODUCTION: The use of dried blood spots (DBS) has gained interest in the field of therapeutic drug monitoring (TDM) due to its potential advantages, such as minimally invasive capillary blood collection, potential stabilization of drugs and metabolites at room or high temperatures, and lower biohazard, allowing for inexpensive storage and transportation. However, there are several drawbacks to the clinical use of DBS in TDM, mostly related to hematocrit (Hct) effects, differences between venous and capillary blood concentrations, among others, that must be evaluated during analytical and clinical method validation. AREA COVERED: This review focuses on the most recent publications on the applications of DBS sampling for TDM (2016-2022), with a special focus on the challenges presented by this alternative sampling strategy, as well as the opportunities for clinical applications. Real-life studies presenting clinical applications were reviewed. EXPERT OPINION: With the availability of method development and validation guidelines for DBS-based methods in TDM, higher levels of assay validation standardization have been achieved, expanding the clinical applications of DBS sampling in patient care. New sampling devices that overcome the limitations of classical DBS, such as the Hct effects, will further encourage the use of DBS in routine TDM.

Evaluation of UGT1A1 and CYP3A Genotyping and Single-Point Irinotecan and Metabolite Concentrations as Predictors of the Occurrence of Adverse Events in Cancer Treatment.

PURPOSE: The variability on irinotecan (IRI) pharmacokinetics and toxicity has been attributed mostly to genetic variations in the UGT1A1 gene, responsible for conjugation of the active metabolite SN-38. Also, CYP3A mediates the formation of inactive oxidative metabolites of IRI. The association between the occurrence of severe adverse events, pharmacokinetics parameters, and UGT1A1 and CYP3A4 predicted phenotypes was evaluated, as the evaluation of [SN-38]/IRI dose ratio as predictor of severe adverse events. METHODS: Forty-one patients undergoing IRI therapy were enrolled in the study. Blood samples were collected 15 min after the end of drug the infusion, for IRI, SN-38, SN-38G, bilirubin concentrations measurements, and UGT1A1 and CYP3A genotype estimation. Data on adverse event was reported. RESULTS: Fifteen patients (36.5%) developed grade 3/4 adverse events. A total of 9.8% (n = 4) of the patients had UGT1A1 reduced activity phenotype, and 48.7% (n = 20) had UGT1A1 and 63.4% (n = 26) CYP3A intermediary phenotypes. Severe neutropenia and diarrhea were more prevalent in patients with reduced UGT1A1 in comparison with functional metabolism (50% and 75% versus 0% and 13%, respectively). SN-38 levels and its concentrations adjusted by IRI dose were significantly correlated to toxicity (rs = 0.31 (p = 0.05) and rs = 0.425 (p < 0.01)). The [SN-38]/IRI dose ratio had a ROC curve of 0.823 (95% CI 0.69-0.956) to detect any severe adverse event and 0.833 (95% CI 0.694-0.973) to detect severe diarrhea. The cut-off of 0.075 ng mL-1 mg-1 had 100% sensitivity and 65.7% specificity to predict severe diarrhea. CONCLUSION: Our data confirmed the relevance of the pre-emptive genotypic information of UGT1A1. The [SN-38]/IRI ratio, measured 15 min after the end of the IRI infusion, was a strong predictor of severe toxicity and could be applied to minimize the burden of patients after IRI administration.

Determination of Ethyl Glucuronide and Ethyl Sulfate in Dried Blood Spots by UHPLC-MS-MS: Method Validation and Assessment of Ethanol Exposure in Postmortem Samples from Road Traffic Victims.

The determination of ethyl glucuronide (EtG) and ethyl sulfate (EtS) in blood has been proposed in clinical and forensic applications to identify recent alcohol consumption. Also, there is a growing interest on the use of dried blood spots (DBS) in toxicological analysis, allowing increased stability of the analytes and simplifying sample transportation and storage. This study presents the development and validation of a method for quantifying EtG and EtS in DBS using ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS-MS). The DBS samples were extracted with a mixture of methanol and acetonitrile (80:20 v/v) and analyzed using UHPLC-MS-MS with electrospray source in negative mode, after separation with a fluoro-phenyl stationary phase. Validation was performed according to the Scientific Working Group for Forensic Toxicology (SWGTOX) guidelines, with calibrations ranging from 0.10 to 18 µg/mL for EtG and 0.02 to 6 µg/mL for EtS. The analytes were stable in DBS stored from -20 to 45°C for 21 days. The method was successfully applied to capillary and venous DBS samples from 20 volunteers after ethanol ingestion and to DBS samples from 99 fatal victims of road traffic injuries. Capillary DBS was comparable to venous DBS and fresh whole blood in Passing-Bablok and Bland-Altman analysis, with correlation coefficients >0.91 (P < 0.001) for all comparisons. In postmortem application, the DBS EtG and EtS analysis indicated positive exposure to ethanol in 72.7% of the cases (EtG: 0.10-24.0 µg/mL and EtS: 0.03-4.11 µg/mL). The identification of ethanol consumption from blood alcohol concentrations (BACs) and EtG/EtS in DBS was in agreement in 98.6% of positive and 96.3% of negative cases (kappa 0.877, P < 0.001), indicating a high level of concordance with BAC in assessing alcohol use in postmortem samples.

Volumetric dried blood microsampling for monitoring imatinib mesylate therapy: Method development and clinical application in patients with chronic myeloid leukemia.

Chronic Myeloid Leukemia (CML) is a hematologic neoplasia, characterized as a proliferative disease of the hematopoietic system. Imatinib mesylate (IM), a selective tyrosine kinase inhibitor, is considered a first-line therapy for CML, indicated for both adult and pediatric patients presenting the Philadelphia chromosome (Ph+). However, patients in treatment with IM may show different responses due to interindividual pharmacokinetic variability. Therapeutic drug monitoring should be routinely performed to identify treatment response profile, adherence to treatment, or possible drug interactions, thus supporting better treatment management. Volumetric absorptive microsampling (VAMS) are innovative devices for blood collection whose advantages include the possibility of home collection by the patient or at the physician's office. The assay was fully validated according to bioanalytical validation guidelines. Estimated plasma concentrations of IM were not statistically different between groups according to adherence (p = 0.15), with median of 789 ng ml-1 in the group with some level of non-adherence versus 1141.9 ng ml-1 in the group with adherence, classified with the Morisky-Green questionnaire. This study included 33 patients with CML in treatment with IM. These patients answered socioeconomic, sociodemographic, and adherence profile (Morisky-Green) questionnaires. Patients also received instructions for home blood collection with VAMS devices. Afterwards, the samples were analyzed by LC-MS/MS. The mean age of the patients was 52 years, 84.8% were ingesting doses of 400 mg/day and the majority were male (69.7%). IM and its metabolite NIM were extracted from VAMS with an aqueous solution with 0.1% formic acid, followed by protein precipitation with acetonitrile. The methodology developed in this study was satisfactory for the determination of IM and NIM in VAMS and can be used in hospital and office routines for the therapeutic monitoring of patients with CML.

Determination of clozapine and norclozapine in dried plasma spot and dried blood spot by liquid chromatography-tandem mass spectrometry.

The use of alternative blood sampling strategies in clozapine (CLZ) therapeutic drug monitoring (TDM) aims to facilitate collection and improve drug therapy and adherence. This study aimed to develop and validate two methods for the determination CLZ and norclozapine (NOR) in dried blood spots (DBS) and dried plasma spots (DPS) by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The analytes were extracted from one 10 microliter volumetric DBS disc punch and from one 6 mm DPS disc punch with methyl tert-butyl ether: methanol (1:9, v/v) and injected into the HPLC-MS/MS with Atmospheric pressure chemical ionization (APCI) source. Separation was performed in a phenyl column, with mobile phase ammonium formate 1 mM pH 4.0 with methanol in gradient mode. The method was linear from 50 to 1500 ng/ml (r > 0.99), with accuracy between 98% and 105% in DBS and 91-101% in DPS, and intra- and inter-assay CV% from 5.23% to 9.35% in DBS and 2.22-11.36% in DPS for both analytes. The matrix effect was compensated by the internal standard, between - 5.1-6.89% in DBS and - 2.45-5.74% in DPS. The average extraction efficiency was 63-67% for CLZ and 58-69% for NOR with no significant impact of hematocrit (HCT). The analytes were stable in the dried matrices stored up to 42 °C for 26 days. The method was applied in the evaluation of clozapine therapy in 13 schizophrenic patients with mean serum levels of 401 ng/ml (43-914 ng/ml). Only 38% were within the therapeutic range, 46% below and 23% above. CLZ and NOR concentrations in dried samples were highly correlated to serum levels, with greater accuracy for DPS compared to DBS (97 versus 89%, and 99 versus 131%, for CLZ and NOR, respectively). Our data support the use of DBS and DPS as alternative sampling strategies in CLZ therapeutic drug monitoring, with satisfactory performance and logistics advantages.

Plasma concentrations of efavirenz are associated with body weight in HIV-positive individuals.

BACKGROUND: Efavirenz is among the most widely used antiretroviral drugs. Increased efavirenz exposure has been associated with CNS side effects and also with the chance of emergence of resistance upon treatment interruptions. The objective of this study was to evaluate factors associated with efavirenz plasma concentrations in a cohort of HIV-infected individuals. METHODS: From July 2009 to March 2010, HIV-infected patients with efavirenz as part of antiretroviral therapy (600 mg at night), undetectable viral load for at least 1 year and CD4 cell count >200 cells/mm(3) were consecutively enrolled at the HIV/AIDS ambulatory care unit in southern Brazil. Plasma samples were taken 18-23 h after efavirenz last dose and analysed by validated ultra-performance liquid chromatography. RESULTS: Forty-one subjects were included (21 females). Mean age and weight were 45.4 years and 70.9 kg, respectively. Mean efavirenz plasma concentration was 2.20 ±â€Š2.17 mg/L. Most plasma concentrations (73%) were within the therapeutic window (1-4 mg/L); 17% were below and 10% above the limits. There were no significant associations between efavirenz concentration and age, CD4 cell count, time on antiretroviral treatment and gender. There was significant and inverse correlation between efavirenz concentrations and body weight (P = 0.013) and body mass index (P = 0.001). CONCLUSIONS: In this cohort of well-controlled HIV-positive individuals, patients with lower weight or body mass index had a higher chance of presenting elevated plasma concentrations of efavirenz. Therapeutic drug monitoring to adjust dose might be a helpful tool to decrease efavirenz dose in order to minimize costs and adverse effects.

Determination of cortisol in hair using liquid chromatography-tandem mass spectrometry: a short review.

Cortisol is considered a particularly relevant biomarker in the context of stress evaluation. This study aims to review of the available literature on the determination of cortisol in hair using LC-MS/MS. Currently, there is no standardized procedure for the measurement of cortisol concentrations in hair, and different sample preparation, chromatographic separation and mass spectrometric detection conditions were described. Simple methanolic extraction, reversed-phase separation and MRM detection in negative ion mode are the most common employed analytical approaches. Reported assays presented acceptable sensitivity for clinical purposes. The increasing use of mass spectrometry in clinical laboratories may contribute to the establishment of LC-MS/MS as the method of choice for the determination of cortisol concentrations in hair.

Sensitive determination of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol and complementary cannabinoids in hair using alkaline digestion and mixed-mode solid phase extraction followed by liquid-chromatography-tandem mass spectrometry.

Hair drug testing can be used for the evaluation of cannabis use with a large detection window, and is required for professional driving license granting in Brazil. A positive hair result for cannabis use requires quantification of the metabolite THC-COOH above the cutoff value of 0.2 ng/g. The achievement of such lower limit of quantification is challenging, particularly with the use of liquid chromatography coupled to triple quadrupole mass spectrometers (LC-MS/MS). In this study, a very sensitive LCMS/ MS assay for the simultaneous quantification of THC-COOH along with THC, CBD, and CBN was developed and validated. Sample preparation was based on hair hydrolysis, followed by selective ion-exchange solid-phase extraction. The extraction yield was 101.5-101.6% for THC-COOH, 92.3-97.4% for THC, 89.7-95.2% for CBN, and 104.9-121.1% for CBD. Internal standard corrected matrix effects were - 2.7 to - 1,1 for THCCOOH and - 11.5 to - 0.1% for the other analytes. The lower limit of quantification was 01 ng/g for THC-COOH and 25 ng/g for THC, CBD, and CBN. The assay fulfilled validation guidelines acceptance criteria. The measurement uncertainties were determined and the assay was ISO17025 accredited, being currently used in routine testing.

Simple determination of valproic acid serum concentrations using BioSPME followed by gas chromatography-mass spectrometric analysis.

Valproic acid (VA) is a drug widely used on the treatment of epilepsy and bipolar affective disorders, with stablished therapeutic concentration ranges in serum. The measurement of VA serum concentrations using chromatographic methods requires a sample preparation step. In this context, this study aims to describe the development and validation of an assay for VA measurement in serum using a new microextraction strategy, known as BioSPME, followed by GC-MS analysis. The extraction procedure was very simple based on direct immersion of the BioSPME tips on acidified serum, followed by agitation and desorption in methanol. The methanolic extracts were directly injected into the chromatograph. Extraction yield was 95.6 to 101.3%. The assay was linear from 10 to 150 mg L-1. Precision, accuracy and stability assays were acceptable according to bioanalytical validation guidelines. The method was applied to 41 clinical serum samples also tested with a previously GC-MS validated assay, which used liquid-liquid extraction as sample preparation. Measurements obtained with both methods were comparable. This study is the first description of the use of BioSPME tips for a therapeutic drug. BioSPME is a promising alternative for the preparation of biological specimens prior to the determination of therapeutic drugs by GC-MS.

Blood phosphatidyl ethanol levels as a tool to detect alcohol misuse in trauma patients.

INTRODUCTION: There is a strong need for a reliable marker of harmful alcohol consumption to identify injured patients that can benefit from alcohol interventions, and blood phosphatidyl ethanol (PEth) has not previously been tested on this population. This study aims to compare the performance of blood PEth concentration, blood alcohol concentration (BAC) and the Alcohol Use Disorders Identification Test Consumption (AUDIT-C) for the screening of alcohol misuse in trauma patients. METHODS: Prospective cross-sectional study of 238 adult patients presenting in the emergency department with any type of trauma. PEth concentration was determined in whole blood by high-performance liquid chromatography with tandem mass spectrometry. Consent, AUDIT-C score and demographic data were obtained. RESULTS: The sample consisted of majority male (67.6%), single (46.2%) and employed (66%) patients. The most common type of trauma was traffic collision (63.9%). The mean age was 41.7 years. We found a significant correlation between PEth levels with AUDIT-C score (Spearman's r = 0.654; p < .0001). PEth had an area under the ROC curve of 0.885 to detect hazardous alcohol consumption (AUDIT-C score ≥ 6) and PEth ≥23.9 ng/mL cutoff point provided 91.2% of sensitivity and 78.4% of specificity. Twelve patients reported alcohol abstinence, but had quantifiable levels of PEth. CONCLUSIONS: PEth levels and AUDIT-C score had a moderate correlation in our population. PEth was useful to identify 12 cases of underreporting of alcohol consumption habits. PEth shows promising results, but more research is needed to identify the best screening tool for alcohol misuse in trauma patients.