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Journal of Pharmaceutical and... Apr 2024The detection of nitazenes in biological fluids is increasingly needed as they are repeatedly reported in intoxication and overdose cases. A simple method for the...
The detection of nitazenes in biological fluids is increasingly needed as they are repeatedly reported in intoxication and overdose cases. A simple method for the quantification of low levels of nine nitazene analogs and brorphine in Dried Blood Spots (DBS) was developed and validated. 10 μL of spiked whole blood is deposited on a Capitainer®B card and allowed to dry. The spot is punched out, and extracted with 500 μL methanol:acetonitrile (3:1 v/v) added with 1.5 μL of fentanyl-D5 as the internal standard. After stirring, sonication, and centrifugation of the vial, the solvent is dried under nitrogen, the extract is reconstituted in 30 μL methanol, and 1 μL is injected into a UHPLC-MS/MS instrument. The method validation showed linear calibration in the 1-50 ng/mL range, LOD values ranging between 0.3 ng/mL (isotonitazene) and 0.5 ng/mL (brorphine), average CV% and bias% within 15 % and 10 % for all compounds, respectively. The matrix effect due to blood and filter paper components was within 85-115 % while recovery was between 15-20 %. Stability tests against time and temperature showed no significant variations for storage periods up to 28 days. Room temperature proved to represent the best samples storage conditions. UHPLC-MS/MS proved capable to reliably identify all target analytes at low concentration even in small specimen volumes, as those obtained from DBS cards, which in turn confirmed to be effective and sustainable micro-sampling devices. This procedure improves the efficiency of toxicological testing and provides an innovative approach for the identification of the nitazene class of illicit compounds.
Topics: Tandem Mass Spectrometry; Methanol; Dried Blood Spot Testing; Chromatography, Liquid; Reproducibility of Results; Chromatography, High Pressure Liquid; Imidazoles; Piperidines
PubMed: 38280237
DOI: 10.1016/j.jpba.2024.115975 -
Analytical and Bioanalytical Chemistry Mar 2024Accurate quantitative analyses require standardized methods to control and improve the analytical process in the laboratory. The availability of urine reference...
Accurate quantitative analyses require standardized methods to control and improve the analytical process in the laboratory. The availability of urine reference materials (RMs) may offer a feasible option to improve the accuracy of urine analysis and to control matrix effects. This paper presents the complete process of the development of matrix RMs in urine, including sample preparation, homogeneity, and stability studies, as well as uncertainty assessment. A freeze-drying process was developed, and freeze-dried human and pig urine samples were prepared and verified to have comparable homogeneity to liquid samples and higher stability than liquid human, pig, and artificial urine samples at 4℃ or room temperature and under extreme conditions. A total of 21 authentic urine samples from August 2022 were measured with freeze-dried RMs and spiked urine samples, and the reliability of the quantification of the RMs was compared. The freeze-dried human urine matrix RM appeared to be an excellent tool for daily quality control, as it showed high stability and gave the most consistent results with spiked samples.
Topics: Humans; Animals; Swine; Reproducibility of Results; Reference Standards; Urinalysis; Specimen Handling; Substance Abuse Detection
PubMed: 38270632
DOI: 10.1007/s00216-024-05142-x -
Analytical Chemistry Feb 2024The accurate and efficient measurement of white blood cell (WBC) counts is vital for monitoring general patient health and can aid in the diagnosis of a range of...
The accurate and efficient measurement of white blood cell (WBC) counts is vital for monitoring general patient health and can aid in the diagnosis of a range of possible infections or diseases. Even with their importance universally acknowledged, access to WBC counts is largely limited to those with access to phlebotomists and centralized clinical laboratories, which house the instruments that perform the tests. As a result, large populations of people (e.g., those that are home-bound or live in remote locations) lack facile access to testing. Dried blood spot (DBS) cards are often used to bridge these gaps in access to testing by offering the ability to collect blood at home for ambient shipping to laboratories. However, it is well understood that these cards, which are prepared from cellulose cardstocks without further modification, suffer from variabilities in accuracy and precision due to uncontrolled sample spreading and hematocrit effects, which have hindered their use to determine WBC counts. In this paper, we present a method to obtain an accurate WBC count using a patterned dried blood spot (pDBS) card, which comprises collection zones that meter volumes of dried blood. Using an input volume of 75 μL of whole blood, we demonstrate that, unlike the gold standard DBS card (Whatman 903), our pDBS design allows for the collection of replicate zones containing a reproducible, average volume of dried blood (12.1 μL, 7.8% CV) over the range of hematocrits from 25 to 55%. We then used qPCR to quantify the gene to determine WBC counts from the volumes of blood that are metered in pDBS zones. We observe that WBC counts generated from our method are comparable to those measured with a HemoCue point-of-care WBC analyzer. Our approach to using pDBS cards as a blood collection device has the potential to support at-home sampling and other patient populations that need WBC counts but lack access to clinical facilities.
Topics: Humans; Hematocrit; Dried Blood Spot Testing; Leukocyte Count; Blood Specimen Collection; Cellulose
PubMed: 38266026
DOI: 10.1021/acs.analchem.3c04439 -
Bioanalysis Feb 2024Clinical dosage of toxic and essential elements in blood is well established and the collection method is still by venipuncture. This method has drawbacks and is not...
Clinical dosage of toxic and essential elements in blood is well established and the collection method is still by venipuncture. This method has drawbacks and is not suited for everyone. Volumetric absorptive microsampling (VAMS) has been shown to have advantages over venipuncture. Using inductively coupled plasma tandem mass spectrometry, a method for quantifying elements in whole blood sampled on VAMS was developed/validated. Method's performance was assessed by comparison with whole blood results. Validation and performance assessment tests tend to show that most of the targeted elements provides accurate and reproducible results comparing to a method of reference. Overall, VAMS presents good preliminary results to eventually become an alternative to venipuncture for blood sampling for some trace elements analysis purposes.
Topics: Tandem Mass Spectrometry; Trace Elements; Blood Specimen Collection; Specimen Handling; Dried Blood Spot Testing
PubMed: 38226838
DOI: 10.4155/bio-2023-0180 -
Expert Review of Anti-infective Therapy 2024Severe acute hepatitis (SAH) is defined by a severe inflammation of hepatocytes in the liver parenchyma which can lead to an acute liver failure, a clinical condition... (Review)
Review
INTRODUCTION
Severe acute hepatitis (SAH) is defined by a severe inflammation of hepatocytes in the liver parenchyma which can lead to an acute liver failure, a clinical condition with high mortality rate that can be triggered by several factors but is usually associated to hepatotropic viruses' infection. In 2022, cases of children with severe acute hepatitis of unknown origin hospitalized in Glasgow, Scotland, were reported. Possible causes of this condition include, but are not limited to, undiagnosed viral (and non-viral) infections, autoimmune hepatitis, drug and/or chemical toxicity, mitochondrial chain respiratory and metabolic disorders.
AREAS COVERED
Herpesviruses can cause severe acute hepatitis, but little is known about the role and the mechanisms of herpesviruses as a causative agent of this type of hepatitis. We review the role of herpesviruses as causative agent of SAH in children and other possible mechanisms involved in this disease.
EXPERT OPINION
Differential diagnosis for herpesvirus in SAH should be implemented in all settings. Alternative fluids, such as saliva and dried blood, could be used in the diagnosis to overwhelm the availability of biological specimens at sufficient volume. In the future, genetic studies could also be added to increase the knowledge about severe acute hepatitis in children.
Topics: Child; Humans; Herpesviridae; Hepatitis; Virus Diseases; Diagnosis, Differential; Acute Disease
PubMed: 38224018
DOI: 10.1080/14787210.2024.2304637 -
Zootaxa Dec 2023Two Portuguese institutions, the Museu Maynense da Academia das Cincias de Lisboa (ACL), and the Museu da Cincia da Universidade de Coimbra (MCUC), house a collection of...
The fish collection of Jos Mariano da Conceio Veloso (17421811) and the beginning of ichthyological research in Brazil, with a taxonomic description of the extant specimens.
Two Portuguese institutions, the Museu Maynense da Academia das Cincias de Lisboa (ACL), and the Museu da Cincia da Universidade de Coimbra (MCUC), house a collection of 85 dried fish specimens prepared in what can be called a fish-herbaria following a process similar to that developed by the Dutch naturalist Johan Frederic Gronovius (16901762). These specimens date back to the late eighteenth century and represent Brazilian taxa. Previous authors assumed that they were part of the collections amassed by the Brazilian-Portuguese naturalist Alexandre Rodrigues Ferreira (17561815) during his philosophical voyage to the Amazon. Here we present a review of these specimens, suggesting that they belonged to Friar Jos Mariano da Conceio Veloso (17421811) and describe the history of dispersal of these collections up the present day. A total of 58 species in 50 genera, 32 families and 19 orders are represented in the collection. Only 8.6% of these specimens represent freshwater species, while 91.4% are marine or brackish water species. The present known distribution of these taxa is focused on southwestern Brazil, which agrees with the area where Veloso collected natural history specimens. A good percentage of the species were undescribed at the time Veloso collected them, and had they been published by him, would have had priority over species described decades later by famous eighteenth and nineteenth century ichthyologists. We also present a brief discussion on the challenges and opportunities of studying historical natural history specimens, with special focus on those amassed during the late eighteenth and early nineteenth century by Brazilian-Portuguese naturalists.
Topics: Humans; Male; Animals; Brazil; Natural History
PubMed: 38220996
DOI: 10.11646/zootaxa.5391.1.1 -
Analytica Chimica Acta Feb 2024The detection and quantification of urinary metabolites play an important role in disease diagnosis. In most cases, urinary analyses are done with liquid urine samples,...
BACKGROUND
The detection and quantification of urinary metabolites play an important role in disease diagnosis. In most cases, urinary analyses are done with liquid urine samples, which must be quickly transported to the laboratory to avoid metabolites degradation that is associated with temperature fluctuations. Consequently, dried sampling devices have emerged to minimize analyte degradation. However, most commercial dried sampling devices are expensive, aggregate low volumes, and need better analytical sensitivity. Therefore, a new dry urine sampling device that is inexpensive, suitable for domestic sampling operation, and efficient for quantifying metabolites without requiring high-resolution instruments is proposed in the present study.
RESULTS
The newly designed dry urine sampling device was produced by 3D printing that efficiently determines 63 urinary organic acids using liquid chromatography coupled with mass spectrometry (LC-MS/MS). The system's efficiency was demonstrated with analytical figures of merit, such as precision, accuracy, and stability of analytes after the sampling and storing of ordinary urine samples. The limits of quantification ranged from 0.01 to 0.42 ng mL-1. Precision and accuracy tests showed relative standard deviations of less than 15 %. The urine stability in the sampling device was high within seven days without any significant degradation of the metabolites. The method was applied to the analysis of 10 human urine samples and compared to a conventional method without the use of the sampling device. The results showed no statistically significant differences, demonstrating the method's efficiency.
SIGNIFICANCE
The proposed 3-D printing device was developed with fast, low-cost manufacturing features and can be manufactured with different volumetric capacities, adaptable to the needs of each user. Furthermore, it is innovative because this is the first sampling device that is effective for the simultaneous storage and preservation of several important urinary metabolites. Thus, it is anticipated that its application would contribute significantly to the identification of metabolic disorders.
Topics: Humans; Liquid Chromatography-Mass Spectrometry; Chromatography, Liquid; Tandem Mass Spectrometry; Specimen Handling; Body Fluids
PubMed: 38220312
DOI: 10.1016/j.aca.2023.342185 -
Anesthesia and Analgesia Feb 2024Fentanyl is widely used for analgesia and sedation in neonates, but pharmacokinetic (PK) analysis in this population has been limited by the relatively large sample... (Observational Study)
Observational Study
Multicenter Population Pharmacokinetics of Fentanyl in Neonatal Surgical Patients Using Dried Blood Spot Specimen Collection Demonstrates Maturation of Elimination Clearance.
BACKGROUND
Fentanyl is widely used for analgesia and sedation in neonates, but pharmacokinetic (PK) analysis in this population has been limited by the relatively large sample volumes required for plasma-based assays.
METHODS
In this multicenter observational study of fentanyl kinetics in neonates up to 42 weeks of postmenstrual age (PMA) who received fentanyl boluses and continuous infusions, dried blood spots were used for small-volume sampling. A population PK analysis was used to describe fentanyl disposition in term and preterm neonates. Covariates for the model parameters, including body weight, PMA, birth status (preterm or term), and presence of congenital cardiac disease, were assessed in a stepwise manner.
RESULTS
Clearance was estimated to be greater than adult clearance of fentanyl and varied with weight. Covariate selection did not yield a significant relationship for age as a continuous or dichotomous variable (term or preterm, the latter defined as birth with PMA of <37 weeks) and clearance.
CONCLUSIONS
A supra-allometric effect on clearance was determined during covariate analyses (exponential scaling factor for body weight >0.75), as has been described in population PK models that account for maturation of intrinsic clearance (here, predominantly hepatic microsomal activity) in addition to scaling for weight, both of which impact clearance in this age group.
Topics: Infant, Newborn; Adult; Humans; Infant; Fentanyl; Pain; Body Weight; Metabolic Clearance Rate; Heart Defects, Congenital
PubMed: 38215717
DOI: 10.1213/ANE.0000000000006808 -
Operative Neurosurgery (Hagerstown, Md.) Jan 2024The endoscopic endonasal transpterygoid approach (TPA), minimally invasive compared with the sublabial transmaxillary and transcranial approaches, still accounts for...
BACKGROUND AND OBJECTIVES
The endoscopic endonasal transpterygoid approach (TPA), minimally invasive compared with the sublabial transmaxillary and transcranial approaches, still accounts for morbidity in benign lateral recess of sphenoid sinus (LRSS) pathologies. Others have suggested an alternative route to the LRSS, the endoscopic contralateral medial transorbital approach (cMTO). However, no quantitative evidence exists to support the clinical application of this approach. This cadaveric study, in a controlled laboratory setting, provides a morphometric comparison of the TPA and cMTO for accessing the LRSS. The study also details the anatomy and technical nuances for optimizing the cMTO corridor.
METHODS
Ten fresh preinjected human cadaveric specimens (20 sides) were dissected with neuronavigation, completing endoscopic cMTO and TPA on each side. Four parameters-working distance to lateral recess, surgical exposure area, angle of attack (AoA), and surgical freedom-were measured for each approach. Relevant osteological measurements in 10 dried human skulls were recorded.
RESULTS
The mean distance from the superior margin of the lacrimal sac impression to the inferior margin of the trochlear fossa was 10.29 ± 1.13 mm, and that from the anterior ethmoidal artery foramina to the posterior lacrimal crest was 9.63 ± 1.23 mm. The mean exposure area around the LRSS was significantly higher in TPA (614.09 ± 40.38 mm2) than in cMTO (391.19 ± 59.01 mm2, P = .001). The mean AoA was 9.83° and 10.24° in the cMTO and TPA, respectively, in the craniocaudal direction (P = .529). In the horizontal plane, it was 9.29° and 10.76° (P = .012). There was no significant difference in surgical freedom between the cMTO and TPA (804.61 and 806.05 mm3, respectively; P = .993).
CONCLUSION
Although comparatively limited exposure area, the cMTO approach has a similar AoA and surgical freedom as TPA and offers better visualization and ergonomic advantages. cMTO provides a feasible, less morbid, multiport technique for benign sphenoid sinus lateral recess pathologies.
PubMed: 38189446
DOI: 10.1227/ons.0000000000001053 -
Is the stability of folates in dried blood microsamples sufficient to perform home-sampling studies?The Analyst Jan 2024Dried blood microsampling is increasingly used for home-sampling and epidemiological studies because of its multiple advantages, including an often greatly improved...
Dried blood microsampling is increasingly used for home-sampling and epidemiological studies because of its multiple advantages, including an often greatly improved analyte stability. However, a critical assessment of the stability under realistic conditions should always be performed as part of the validation, especially for unstable molecules like folates (vitamin B9). Here, the objective was to determine whether folate stability in dried blood microsamples is sufficient to allow the set-up of home-sampling studies for the monitoring of folate status in , women of reproductive age. An extensive set of stability experiments was performed to evaluate the stability of the main folate vitamer 5-methyltetrahydrofolate (5MTHF), its oxidation product MeFOX and the minor non-methyl folate vitamers 10-formylfolic acid (10FoFA), 5,10-methenyltetrahydrofolate (5,10CH+THF) and tetrahydrofolate (THF) in dried blood microsamples using volumetric absorptive microsampling (VAMS) or regular dried blood spots (DBS). The evaluations included (EDTA-anticoagulated blood was collected from a single donor measured in four replicates per condition and time point): (i) the effect of temperature (-20 °C, 4 °C, ambient temperature and 37 °C), (ii) the effect of light (during drying and storage) and humidity, and (iii) the effect of storage under vacuum and pretreatment of the microsamples with stabilizing agents on folate stability. At -20 °C and 4 °C, all folate levels were within 85 to 115% of the baseline value up till two weeks of storage in both VAMS samples and DBS. However, at room temperature the stability of the analyzed folates was only consistently observed up till three days in VAMS samples, and for none of the folates at 37 °C. Humidity had a major impact on 5,10CH+THF stability, but this could be easily improved by using desiccant. Both vacuum treatment and pretreatment of microsamples with 0.1% DL-dithiothreitol and 5% butylated hydroxytoluene improved the stability at room temperature in VAMS samples, but these effects were limited at 37 °C and in DBS. Overall, the stability of the individual folate vitamers proved to be challenging and strongly temperature- and time-dependent. Nonetheless, if controlled transport (temperature and duration) can be assured, the set-up of home-sampling studies to evaluate the folate status using dried blood microsamples can still be beneficial.
Topics: Humans; Female; Dried Blood Spot Testing; Sampling Studies; Tandem Mass Spectrometry; Specimen Handling; Temperature
PubMed: 38189100
DOI: 10.1039/d3an01004j