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Apixaban, a direct oral anticoagulant drug (DOAC), typically does not require routine therapeutic drug monitoring (TDM), yet recent guidelines propose its use in specific clinical scenarios. While various antifactor Xa (anti‐FXa) chromogenic assays serve as useful proxies for measuring plasma exposure to apixaban in emergencies, they lack specificity compared with chromatographic methods. This research project is intended to the development and validation of a standardized protocol of liquid chromatography–tandem mass spectrometry (LC–MS/MS) in conformity with the ICH guidelines M10 for the measurement of apixaban in both plasma and dried plasma spots (DPSs). Samples preparation included protein precipitation after the addition of a deuterated internal standard (IS), and the chromatographic separation was carried out on a Thermo Scientific™ Accucore™ Polar Premium column (50 mm × 2.1 mm, i.d. 2.6 m). The newly developed LC–MS/MS method for apixaban mesurement from both plasma and DPS resulted linear over a wide concentration range (31.25–500 ng/mL), accurate, and reproducible without matrix effects, allowing for specific and rapid quantification. Stability was assessed on quality controls and a real sample, allowing the setting up of a robust TDM protocol that was applied to five anonymized plasma samples obtained from adult patients undergoing apixaban treatment at steady‐state. In conclusion our novel LC–MS/MS method is adequate for accurate apixaban quantitation from both plasma and DPS matrixes, and may thus facilitate the guidelines suggested implementation of apixaban TDM, even in peripheral hospitals through shipment of DPS at reference laboratories. [ABSTRACT FROM AUTHOR]

Aims: Therapeutic drug monitoring (TDM) has led to significant improvements in individualized medical care, although its implementation in oncology has been limited to date. Tyrosine kinase inhibitors (TKIs) are a group of therapies for which TDM has been suggested. Osimertinib is one such therapy used in the treatment of epidermal growth factor receptor (EGFR) mutation‐driven lung cancer. Herein, we describe a prospective pilot study involving 21 patients on osimertinib primarily as a preliminary evaluation of drug levels in a real‐world setting. Methods: Concentrations of the drug and its primary metabolites were measured with a validated liquid chromatography–mass spectrometry (LC–MS) assay across serial timepoints. As part of this study, inter‐individual variability by dose and ethnicity as well as intra‐individual variability across timepoints are explored. Furthermore, we attempted to validate dried blood spot (DBS)‐based quantitation as an accurate alternative to plasma quantitation. Results: Successful quantitation of osimertinib and primary metabolites was achieved for our subjects. Compound plasma levels were highly correlated to DBS levels. There was no significant difference in concentrations with ethnicity or dosing or intra‐individual variability across timepoints. Conclusions: As such, we demonstrate that TDM for osimertinib is practical for future trials. We also validated the use of DBS as an alternative to conventional quantitation for exploration of TDM for osimertinib in larger trials and for other targeted therapies. [ABSTRACT FROM AUTHOR]

Microsampling has revolutionized pharmaceutical drug development and clinical research by reducing sample volume requirements, allowing sample collection at home or nontraditional sites, minimizing animal and patient burden, and enabling more flexible study designs. This perspective paper discusses the transformative impact of microsampling and patient‐centric sampling (PCS) techniques, emphasizing their advantages in drug development and clinical trials. We highlight the integration of liquid chromatography‐mass spectrometry (LC–MS) strategies for analyzing PCS samples, focusing on our research experience and a review of current literatures. The paper reviews commercially available PCS devices, their regulatory status, and their application in clinical trials, underscoring the benefits of PCS in expanding patient enrollment diversity and improving study designs. We also address the operational challenges of implementing PCS, including the need for bridging studies to ensure data comparability between traditional and microsampling methods, and the analytical challenges posed by PCS samples. The paper proposes future directions for PCS, including the development of global regulatory standards, technological advancements to enhance user experience, the increased concern of sustainability and patient data privacy, and the integration of PCS with other technologies for improved performance in drug development and clinical studies. By advancing microsampling and PCS techniques, we aim to foster patient‐centric approaches in pharmaceutical sciences, ultimately enhancing patient care and treatment efficacy. [ABSTRACT FROM AUTHOR]

This work aimed to simplify and improve the process of binding monoclonal antibodies (mAbs) covalently to filter paper for use in dried blood spot sampling, enabling instant capture of protein biomarkers for targeted protein determination. Incorporating the necessary immunocapture sample preparation step in the initial sampling stage saves time and reduces the workload. The biomarker human chorionic gonadotropin (hCG) was used as the model analyte. The antibody‐based paper samplers were prepared by functionalizing paper discs (6 mm) through a simple reaction using divinyl sulfone (DVS). After DVS activation, the paper discs were incubated with E27 hCG mAbs, followed by 0.05% tween/phosphate buffer saline to block the surface. After sample application and drying, the discs only needed to be washed before tryptic digestion and finally analysed on a nanoliquid chromatography–tandem mass spectrometry system. The finished DVS‐mAbs samplers could selectively capture hCG (100 ng/mL) from human serum, with a recovery of 50%. Sample clean‐up reduced the number of identified proteins from 132 to 82 before and after wash, respectively, with a 70% reduction in serum albumin signal while still retaining hCG on the sampler during the washing protocol. An evaluation of the samplers revealed excellent linearity (R2 = 0.9995) for hCG in serum with relative standard deviations below 15%. This work has presented the first ever reported paper samplers immobilized with antibodies utilizing DVS chemistry, showing promise in the future of paper‐based sampling. [ABSTRACT FROM AUTHOR]

Introduction: The offspring of CD‐1 mice exposed during pregnancy to one of seven gadolinium‐based contrast agents (GBCAs) were evaluated for potential effects on postnatal development and behavior. The GBCAs, comprising four linear (gadopentetate dimeglumine, gadodiamide, gadobenate dimeglumine, and gadoxetate disodium) and three macrocyclic (gadoterate meglumine, gadoteridol, and gadobutrol), were administered via intravenous injection once daily from Gestation Day 6 through 17 following confirmed mating (Day 0) at doses of at least twice the human equivalent recommended clinical dose (i.e., 0.63 mmol Gd/kg for gadoxetate disodium and 2.5 mmol Gd/kg for the other GBCAs). All dams were allowed to deliver naturally. F0 generation females were monitored for maternal toxicity and gadolinium (Gd) levels in blood and brain. Offspring were evaluated for Gd levels in blood and brain at birth and on Day 70 postpartum. F1 generation mice were evaluated for survival and growth preweaning. Selected pups/litter were evaluated postweaning for sexual maturation, growth, and behavior. Gd was quantifiable in the brain of the F1 offspring on PND 1, with levels declining over time. There was no long‐term effect of any GBCA on the growth and development of any offspring. There was no impact on neurodevelopment, as assessed by brain histology and validated neurobehavioral tests, including a battery of functional observational tests, motor activity, and learning and memory as evaluated in the Morris water maze. Conclusion: At the end of the postweaning period, the highest dose tested was considered the no‐observable‐adverse‐effect level (NOAEL) in the F0 and F1 offspring for all tested GBCAs. [ABSTRACT FROM AUTHOR]

Introduction: Seven gadolinium‐based contrast agents (GBCAs), four linear and three macrocyclic, were evaluated for potential effects on development, including behavior of juvenile CD‐1 mice. Methods: The GBCAs were administered via intravenous injection once daily on postnatal day (PND) 9, 12, 15, 18, and 21 (PND 1 was the day of delivery) at doses up to twice the human equivalent clinical dose (i.e., 0.63 mmol Gd/kg for gadoxetate disodium and 2.5 mmol Gd/kg for the other GBCAs). Mice were bled for evaluation of exposure (plasma) to gadolinium (Gd) on PND 9, 12, and 70. At scheduled euthanasia, the liver, spleen, brain, skin (dorsal surface), bone (left femur), and kidneys were excised from up to six mice/sex/group on PND 10, 22, or 70 for the determination of Gd levels and histopathological analysis. All mice were monitored for toxicity, growth and survival, sexual maturation, and behavior. Conclusion: Gd was quantifiable in the brain tissues with levels declining over time. There was no long‐term effect on the growth and development for mice exposed to any of the GBCAs. There was no impact on neurodevelopment as assessed by brain histology and validated neurobehavioral tests, including a functional observational battery, motor activity, and learning and memory as evaluated in the Morris water maze. For all GBCAs, the highest dose tested represented the no‐observable‐adverse‐effect level in juvenile mice. [ABSTRACT FROM AUTHOR]

Recently, animal welfare has been attracting worldwide attention, and implementation of 3Rs (replacement, reduction, and refinement) is prioritized in every way possible in the drug development. Microsampling, in which small amounts of blood are collected, is attracting attention in this context. ICH S3A Q&A focused on microsampling was published in November 2017 to help accelerate the application of microsampling for toxicokinetic assessment. The increased sensitivity of drug measurement apparatuses such as mass spectrometers has made it possible to measure drug concentrations with small amounts of blood samples. In this review, we summarized the reports on toxicological influence of microsampling in rodents (rats and mice) with or without drug administration or recovery period after blood collection and influences that may arise from differences in the blood sampling site or blood sampling volume. We also summarized some perspectives on further implementation of microsampling in toxicology studies. The use of microsampling in regulatory toxicology studies has gradually increased, although at a lower rate than in discovery studies. Since more animals are used in GLP toxicology studies than in discovery studies, the effect of reducing the number of animals by microsampling is expected to be greater in the toxicology studies. This report aims to promote the application of microsampling to nonclinical studies, as it is beneficial for improving animal welfare and can contribute to the 3Rs. Microsampling is a technique for collecting small amounts of blood from animals and is expected to contribute to the 3Rs. In this review, we summarized the reports on toxicological influence of microsampling in rodents (rats and mice) with or without drug administration or recovery period after blood collection and influences that may arise from differences in the blood sampling site or blood sampling volume. We also summarized some perspectives on further implementation of microsampling in regulatory toxicology studies. [ABSTRACT FROM AUTHOR]

This perspective explores the feasibility of smart sampling with dried blood spots for the determination of proteins and peptides from human biomatrices using liquid chromatography coupled to mass spectrometry for clinical purposes. The focus is on innovative approaches to transform filter paper from a mere sample carrier to an active element in sample preparation, with the aim of reducing the need for extensive and intensive sample preparation in the conventional sense. Specifically, we discuss the use of modified cellulose to integrate sample preparation at an early stage of sample handling. The use of paper immobilized with either trypsin or monoclonal antibodies for protein digestion and affinity clean‐up is discussed as a potential benefit of starting sample preparation instantly at the moment of sampling to optimize time efficiency and enable faster analysis, diagnosis, and follow‐up of patients. [ABSTRACT FROM AUTHOR]

With the development of highly sensitive bioanalytical techniques, the volume of samples necessary for accurate analysis has reduced. Microsampling, the process of obtaining small amounts of blood, has thus gained popularity as it offers minimal‐invasiveness, reduced logistical costs and biohazard risks while simultaneously showing increased sample stability and a potential for the decentralization of the approach and at‐home self‐sampling. Although the benefits of microsampling have been recognised, its adoption in clinical practice has been slow. Several microsampling technologies and devices are currently available and employed in research studies for various biomedical applications. This review provides an overview of the state‐of‐the‐art in microsampling technology with a focus on the latest developments and advancements in the field of microsampling. Research published in the year 2022, including studies (i) developing strategies for the quantitation of analytes in microsamples and (ii) bridging and comparing the interchangeability between matrices and choice of technology for a given application, is reviewed to assess the advantages, challenges and limitations of the current state of microsampling. Successful implementation of microsampling in routine clinical care requires continued efforts for standardization and harmonization. Microsampling has been shown to facilitate data‐rich studies and a patient‐centric approach to healthcare and is foreseen to play a central role in the future digital revolution of healthcare through continuous monitoring to improve the quality of life. [ABSTRACT FROM AUTHOR]

In the wake of the COVID‐19 global pandemic, self‐administered microsampling tools have reemerged as an effective means to maintain routine healthcare assessments without inundating hospitals or clinics. Finger‐stick collection of blood is easily performed at home, in the workplace, or at the point‐of‐care, obviating the need for a trained phlebotomist. While the initial collection of blood is facile, the diagnostic or clinical utility of the sample is dependent on how the sample is processed and stored prior to transport to an analytical laboratory. The past decade has seen incredible innovation for the development of new materials and technologies to collect low‐volume samples of blood with excellent precision that operate independently of the hematocrit effect. The final application of that blood (i.e., the test to be performed) ultimately dictates the collection and storage approach as certain materials or chemical reagents can render a sample diagnostically useless. Consequently, there is not a single microsampling tool that is capable of addressing every clinical need at this time. In this review, we highlight technologies designed for patient‐centric microsampling blood at the point‐of‐care and discuss their utility for quantitative sampling as a function of collection material and technique. In addition to surveying methods for collecting and storing whole blood, we emphasize the need for direct separation of the cellular and liquid components of blood to produce cell‐free plasma to expand clinical utility. Integrating advanced functionality while maintaining simple user operation presents a viable means of revolutionizing self‐administered microsampling, establishing new avenues for innovation in materials science, and expanding access to healthcare. [ABSTRACT FROM AUTHOR]

Microsampling technology for dried blood–derived samples provides an advantageous alternative to conventional venous blood for drug quantitation. Unlike conventional whole blood microsampling techniques, Noviplex is a novel, card‐based technology for rapid dried plasma spot collection that retains the benefits of microsampling during collection and transportation, while avoiding the disadvantages of using whole blood samples. Giredestrant is a promising small‐molecule therapeutic agent under development by Genentech to treat patients with estrogen receptor–positive breast cancer. In this study, we investigated the feasibility of using Noviplex cards for pharmacokinetic analysis of giredestrant levels in human plasma, including optimizing extraction recovery, evaluating in‐card stability, and assessing batch precision and accuracy. We found that while the Noviplex card demonstrated levels of sensitivity, extraction recovery, and stability at ambient temperature that meet the requirements of pharmacokinetic analysis for clinical studies, further optimization of the filtration layers within the Noviplex card is necessary to improve filtration efficiency and consistency. This study reveals the possibilities as well as the limitations of the Noviplex card and provides a better understanding of the capabilities and risks of using the Noviplex card for drug quantitation in plasma. [ABSTRACT FROM AUTHOR]

The limitations of many in situ microsampling techniques, especially when applied to isotopic analyses, may result in lower measurement precision and limit the ability to compare different isotope systems within the same phase or the same isotope system between phases. High‐precision, combined and complementary isotopic measurements are typically limited to whole‐rock samples or mineral separates at the expense of textural context. We demonstrate that laser ablation instruments could offer an effective, non‐mechanical alternative to micro‐milling for delineating and extracting single crystals, interstitial phases, and sub‐crystal scale features from soluble‐resin thick sections for column chromatography and high precision ID‐TIMS measurement. Systematic testing demonstrated that both modern and legacy laser instruments are capable of ablating through plagioclase, pyroxene and fused glass to depths of 200 μm. Modern instruments offer software benefits to identifying optically‐indistinct petrographic features, chemical zoning and avoiding alteration phases by using overlain high‐resolution datasets. Extracting delineated targets of varying geometry was achieved under binocular microscope using common mineral‐separation tools. Analysis of RM glass used as mineral proxies indicate (a) no significant thermal aureole on ablated margins that could affect measured parent‐daughter ratios, (b) no elevated risk of contamination compared with traditional mineral separation techniques, (c) excellent blanks for low‐yield analyses and (d) high‐precision ID‐TIMS results (87Sr/86Sr and 143Nd/144Nd) for sub‐100 ng Sr loads and sub‐10 ng Nd loads. [ABSTRACT FROM AUTHOR]

Aims: Immunosuppressant and kidney function monitoring are crucial for kidney transplant recipient follow‐up. Microsamples enable remote sampling and minimise patient burden as compared to conventional venous sampling at the clinic. We developed a liquid chromatography–tandem mass spectrometry assay to quantify tacrolimus, mycophenolic acid (MPA), creatinine and iohexol in dried blood spot (DBS), and volumetric absorptive microsample (VAMS) samples. Methods: The assay was successfully validated analytically for all analytes. Clinical validation was conducted by direct comparison of paired DBS, VAMS and venous reference samples from 25 kidney transplant recipients. Patients received iohexol 5–15 minutes before immunosuppressant intake and were sampled 0, 1, 2 and 3 hours thereafter, enabling tacrolimus and MPA area under the concentration–time curve (AUC) and creatinine‐based and iohexol‐based glomerular filtration rate (GFR) estimation. Method agreement was evaluated using Passing–Bablok regression, Bland–Altman analysis and the percentages of values within 15–30% of the reference (P15–P30) with a P20 acceptance threshold of 80%. Results: For DBS samples, method agreement was excellent for tacrolimus trough concentrations (n = 25, P15 = 92.0%) and AUCs (n = 25; P20 = 95.8%) and adequate for creatinine‐based GFR trend monitoring (n = 25; P20 = 80%). DBS‐based MPA AUC assessment showed suboptimal agreement (n = 16; P20 = 68.8%), but was considered acceptable given its P30 of 100%. The assay performed inadequately for DBS‐based iohexol GFR determination (n = 24; P20 = 75%). The VAMS technique generally showed inferior performance, but can be considered for certain situations. Conclusion: The assay was successfully validated for tacrolimus, MPA and creatinine quantification in DBS samples, enabling simultaneous remote kidney function trend monitoring and immunosuppressant therapeutic drug monitoring in kidney transplant recipients. [ABSTRACT FROM AUTHOR]

Phosphatidylethanols (PEth) are a group of abnormal phospholipids which are formed within the human body in the presence of ethanol. As PEth accumulates in red blood cells, which prolongs its detectability, it became popular as a direct alcohol biomarker. When alcohol is consumed regularly, PEth concentrations in the human blood reach an equilibrium (plateau) between formation and degradation which is representative of drinking habits. A wide range of applications has been demonstrated for PEth, whereby in most studies it could provide advancements regarding accuracy and precision when compared to already established alcohol biomarkers (carbohydrate‐deficient transferrin, gamma‐glutamyltransferase, and ethyl glucuronide). Although PEth has been the focus of many research projects all around the world, the final breakthrough from the research laboratory into the clinical mass market is still ongoing. In this overview article, we discuss the evolution of PEth analysis, its applications, and the associated challenges. The main reasons that hinder the use of PEth in the mass market were identified as (a) the lack of implementing a uniform reference measurement system, which should ensure that every single PEth result is comparable and/or (b) ensuring good laboratory practice dedicated to PEth analysis, which should guarantee that every result obtained by an individual laboratory is correct. These challenges regarding PEth analytics must be carefully addressed in the near future before the implementation of common cut‐off values and international guidelines can take place. This article is categorized under:Toxicology > AlcoholToxicology > Analytical ToxicologyToxicology > Drug Analysis [ABSTRACT FROM AUTHOR]

Here the relation between digestion of proteins by trypsin covalently bound to paper and trypsin in‐solution is investigated. The trypsin acting on paper is covalently bound. A trypsin concentration of 0.5% (w/v) results in the highest digestion activity of all concentrations tested. Additionally, it can be seen that trypsin on‐paper has retained approx. 50% of its activity. Unlike trypsin in‐solution, the stability of the smart proteolysis samplers was regarded to be stable for at least four months when kept refrigerated. Autolysis was very small for covalently bound trypsin: less than 2% compared to in‐solution trypsin. Proteomic analysis of diluted human serum showed more protein identifications (214) in‐solution digestions than on‐paper digestions (76). Also, higher coverage for the in‐solution digestion was obtained. Those proteins identified after on‐paper digestion with no or few disulfide bonds seem to have more similar sequence coverages compared to those identified after in‐solution digestion. Smart samplers allow the determination of at least 70–75 proteins without performing the overnight digestion. All in all, trypsin covalently bound to paper shows to retain high proteolytic activity and is a stable alternative for conventional digestions. In this way, smart proteolytic samplers show their feasibility in pre‐lab sample preparation. [ABSTRACT FROM AUTHOR]

Epilepsy is one of the most common diseases of the central nervous system globally. To ensure the correct dosage of antiepileptic treatment, it is helpful to check the blood levels of the administered substances regularly. The analysis of the capillary dried blood samples provides a promising and less‐invasive alternative to venous blood collection. Therefore, the aim of the present study was to develop an LC–MS method for the quantification of 22 commonly used drugs in patients with an epileptic syndrome and 5 drug metabolites in one dried blood spot (DBS). The calibration ranges were selected in such a way that the therapeutic reference ranges in serum for the respective substances were completely covered. The analytical validation was successfully performed according to relevant guidelines with a consideration of requirements for DBS analysis. Proof of concept of the developed method was obtained by the analysis of DBSs from 282 authentic leftover ethylenediaminetetraacetic acid blood samples, which were compared with the corresponding serum concentrations. Altogether, the results show a dependency on the blood/plasma (b/p) ratios of the respective analytes so that for drugs with b/p ratios close to one, for example, lacosamide, levetiracetam, brivaracetam, and sertraline, a good accordance was observed. [ABSTRACT FROM AUTHOR]

The outbreak of the new coronavirus disease changed the world upside down. Every day, millions of people were subjected to diagnostic testing for Covid‐19, all over the world. Molecular tests helped in the diagnosis of current infection by detecting the presence of viral genome whereas serological tests helped in detecting the presence of antibody in blood as well as contributed to vaccine development. This testing helped in understanding the immunogenicity, community prevalence, geographical spread and conditions post‐infection. However, with the contagious nature of the virus, biological specimen sampling involved the risk of transmission and spread of infection. Clinic or pathology visit was the most concerning part. Trained personnel and resources was another barrier. In this scenario, microsampling played an important role due to its most important advantage of remote, contactless, small volume and self‐sampling. Minimum requirements for sample storage and ease of shipment added value in this situation. The highly sensitive instruments and validated assay formats assured the accuracy of results and stability of samples. Microsampling techniques are contributing effectively to the Covid‐19 pandemic by reducing the demand for clinical staff in population‐level testing. The validated and established applications supported the use of microsampling in diagnosis, therapeutic drug monitoring, development of treatment or vaccines and clinical trials for Covid‐19. [ABSTRACT FROM AUTHOR]

In the current study, two groups of rats (five per group) were administered a single oral dose of 500 mg/kg acetaminophen. For toxicokinetic assessment, the Group 1 animals were bled via conventional sparse (two animals/time point) sublingual vein bleeding (~0.5 ml) with anesthesia, while the Group 2 animals were bled via serial tail vein microsampling (~0.075 ml) without anesthesia. All collected blood was processed for plasma. Each Group 2 plasma sample (~30 μl) was divided into 'wet' and 'dried' (dried plasma spots). All plasma samples were analyzed by LC–MS/MS for acetaminophen and its major metabolites acetaminophen glucuronide and acetaminophen sulfate. In addition, plasma and urine samples were collected for analysis of corticosterone and creatinine to assess stress levels. Comparable plasma exposure to acetaminophen and its two metabolites was observed in the plasma obtained via conventional sparse sublingual vein bleeding and serial tail vein microsampling and between the 'wet' and 'dried' plasma obtained by the latter. Furthermore, comparable corticosterone levels or corticosterone/creatinine ratios between the two groups suggested that serial microsampling without anesthesia did not increase the levels of stress as compared with conventional sampling with anesthesia, confirming the utility of microsampling for plasma or dried plasma spots in rodent toxicokinetic assessment. [ABSTRACT FROM AUTHOR]

Recent advancements in the sensitivity of chemical instrumentation have led to increased interest in the use of microsamples for translational and biomedical research. Paper substrates are by far the most widely used media for biofluid collection, and mass spectrometry is the preferred method of analysis of the resultant dried blood spot (DBS) samples. Although there have been a variety of review papers published on DBS, there has been no attempt to unify the century old DBS methodology with modern applications utilizing modified paper and paper‐based microfluidics for sampling, storage, processing, and analysis. This critical review will discuss how mass spectrometry has expanded the utility of paper substrates from sample collection and storage, to direct complex mixture analysis to on‐surface reaction monitoring. [ABSTRACT FROM AUTHOR]

Abstract: This study presents Lu–Hf geochronology of zoned garnet in high‐P eclogites from the North Qilian orogenic belt. Selected samples have ~mm‐sized garnet grains that have been sampled with a micro‐drill and analysed for dating. The Lu–Hf dates of bulk garnet separates, micro‐drilled garnet cores and the remnant, rim‐enriched garnet were determined by two‐point isochrons, with cores being consistently older than the bulk‐ and rim‐enriched garnet. The bulk garnet separates of each sample define identical garnet–whole rock isochron date of c. 457 Ma. Consistent U–Pb zircon dates of 455 ± 8 Ma were obtained from the eclogite. The Lu–Hf dates of the drilled cores and rim‐rich separates suggest a minimum garnet growth interval of 468.9 ± 2.4 and 452.1 ± 1.6 Ma. Major and Lu element profiles in the majority of garnet grains show well‐preserved Rayleigh‐style fractionated bell‐shaped Mn and Lu zoning profiles, and increasing Mg from core to rim. Pseudosection modelling indicates that garnet grew along a P–T path from ~470–525°C and ~2.4–2.6 GPa. The exceptional high‐Mn garnet core in one sample indicates an early growth during epidote–blueschist facies metamorphism at <460°C and <0.8 GPa. Therefore, the Lu–Hf dates of drilled cores record the early prograde garnet growth, whereas the Lu–Hf dates of rim‐rich fractions provide a maximum age for the end of garnet growth. The microsampling approach applied in this study can be broadly used in garnet‐bearing rocks, even those without extremely large garnet crystals, in an attempt to retrieve the early metamorphic timing recorded in older garnet cores. Given a proper selection of the drill bit size and a detailed crystal size distribution analysis, the cores of the mm‐sized garnet in most metamorphic rocks can be dated to yield critical constraints on the early timing of metamorphism. This study provides new crucial constraints on the timing of the initial subduction (before c. 469 Ma) and the ultimate closure (earlier than c. 452 Ma) of the fossil Qilian oceanic basin. [ABSTRACT FROM AUTHOR]

Abstract: Compounds and products in the biocide and plant protection sector can only be registered after formal risk assessment to ensure safety for users and the environment. In bird and mammal risk assessment, this is routinely done using generic focal species as models, which are of particular exposure risk. Such a species is the common vole (Microtus arvalis) due to its high food intake relative to the low body weight. For wild species, biological samples, data and hence realistic exposure estimations are particularly difficult to obtain. In recent years, advances have been made in the techniques related to serial microsampling of laboratory mice and rats that allow for a reduction in sampling volumes. Similar progress in wild species sampling is missing. This study presents a proof of concept to dose wild rodents with relevant compounds and to draw serial, low volume blood samples suitable for state‐of‐the art toxicokinetic analyses. For the first time, the jugular vein of common voles was used to administer compounds (two frequently used fungicidal components). This procedure and the following microsampling of blood (2 × 10 μl six times within 24 hours) from the lateral tail vein did not affect body weight and mortality of voles. Samples were sufficient to detect dissipation patterns of the compounds from blood in toxicokinetic analysis. These results suggest that microsampling can be well translated from laboratory mice to wild rodent species and help to obtain realistic exposure estimates in wild rodents for ecotoxicological studies as well as to promote the 3R concept in studies with wild rodent species. [ABSTRACT FROM AUTHOR]

In preclinical safety assessment, blood volume requirements for various endpoints pose a major challenge. The goal of this working group was to review current practices for clinical pathology (CP) testing in preclinical toxicologic studies, and to discuss advantages and disadvantages of methods for reducing blood volume requirements. An industry-wide survey was conducted to gather information on CP instrumentation and blood collection practices for hematology, clinical biochemistry, and coagulation evaluation in laboratory animals involved in preclinical studies. Based on the survey results and collective experience of the authors, the working group proposes the following 'points to consider' for CP testing: (1) For most commercial analyzers, 0.5 mL and 0.8 mL of whole blood are sufficient for hematology and biochemistry evaluation, respectively. (2) Small analyzers with low volume requirements and low throughput have limited utility in preclinical studies. (3) Sample pooling or dilution is inappropriate for many CP methods. (4) Appropriate collection sites should be determined based on blood volume requirements and technical expertise. (5) Microsampling does not provide sufficient volume given current analyzer and quality assurance requirements. (6) Study design considerations include: the use of older/larger animals (rodents), collection of CP samples before toxicokinetic samples, use of separate subsets of mice for hematology and clinical biochemistry testing, use of a priority list for clinical biochemistry, and when possible, eliminating coagulation testing. [ABSTRACT FROM AUTHOR]

Background Clinical pathology of rodents is hindered by sample volume limitations. A single whole heparinized blood sample is often submitted for hematologic and clinical chemistry analysis in exploratory research settings, and sample dilution may be required. Published information on the potential impact of sample dilution and heparin use on hematology variables in rodents is sparse. Objectives The purpose of the study was to evaluate the effects of sample dilution and of anticoagulant on hematologic analysis of mouse and rat blood samples on the Sysmex XT-2000iV. Methods Mice and rats were obtained from various ends of study research projects, and whole blood was collected via terminal cardiocentesis in lithium heparin, and additionally in EDTA when paired samples were obtained from rats. Hematology analytes were measured on the Sysmex XT-2000iV straight and diluted from ×2 to ×5. Results Significant differences between heparinized samples analyzed straight vs diluted were found for MCV and MCHC, with a bias for several additional variables. Significant differences between paired heparinized and EDTA-anticoagulated samples at each dilution point were found for most variables, with the largest differences found in platelet count. Evidence of platelet clumping presumably due to heparin exposure was noted in numerous samples. Conclusions Dilution-induced changes occur in some hematologic variables and may render dilution unacceptable in the exploratory research environment. Many variables, most notably platelet count, differ based on the anticoagulant used, and values from heparinized vs EDTA-anticoagulated samples should not be directly compared. [ABSTRACT FROM AUTHOR]

ABSTRACT This article evaluates age of weaning and early childhood diets of eight males and nine females from a Middle Holocene (4300-3000 BP) site in Central California, CA-CCO-548. All individuals died as adults. δ15N values from serial sections of dentin collagen in first molars suggest females were fully weaned, on average, by 3.6 years of age, about 0.4 years later than males in the sample, suggesting possible greater parental investment in female offspring. However, throughout childhood females consumed lower trophic-level foods than males. This could indicate greater investment in males through provisioning of higher quality foods, or alternatively, some degree of independent foraging by males starting as early as 2 to 3 years of age. Even as adults, these same males and females consumed a different range of foods as indicated by their bone collagen δ13C and δ15N values. Overall, the data suggest children were enculturated early into their respective gendered diets, with girls consuming greater amounts of plant foods and boys consuming greater amounts of higher-trophic level fish and meat protein. Am J Phys Anthropol 152:471-483, 2013. © 2013 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

A novel sampling technique for the analysis of glass is described, which involves the removal of minute particles from an object with a diamond-coated file, followed by energy-dispersive X-ray analysis in the scanning electron microscope (SEM/EDXA). The particles are fixed to adhesive carbon discs and carbon coated without grinding or polishing. Mean compositions are determined for 10 arbitrarily selected particles above a minimum grain size of 150 mm and normalized to totals of 100%. Tests were carried out on two standard soda–lime–silica glasses of well-characterized composition, using two files of different grade. The analyses showed good agreement with the accepted values of all elements. Although the precision is somewhat reduced, this highly portable and quasi non-destructive microsampling procedure provides almost the same information as that gained from samples embedded and polished in the normal way. Its application is thought to be especially useful for the investigation of intact glass objects and ceramic glazes. [ABSTRACT FROM AUTHOR]