Alternative Sampling Strategies Research Articles

A novel approach to therapeutic drug monitoring of Ciclosporin in pediatric renal transplant recipients using volumetric absorptive microsampling (VAMS) – Teaching old dog new tricks

Background and aimsCiclosporin (CSA) is an immunosuppressive agent that requires therapeutic drug monitoring (TDM). High partitioning in erythrocytes indicates that whole blood (WB) is a suitable matrix for CSA determination. Alternative sampling strategies, such as volumetric absorptive microsampling (VAMS), are novel possibilities for blood collection during TDM for various analytes, including immunosuppressants. This technique is attractive for vulnerable pediatric patients, including home-based self-sampling, remote therapy, and adherence control. Materials and methodsThis study aimed to develop and validate a new method for CSA determination based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) of WB and VAMS samples. Additionally, these methods were applied for CSA determination in clinical samples from pediatric transplant recipients. A strong point of this study is the assessment of an external proficiency testing scheme. ResultsBoth methods were successfully validated within the 1–2000 ng/mL calibration range, with LOD 0.5 and 1 ng/mL for WB and VAMS methods, respectively. All the validation parameters fulfilled the international acceptance criteria for bioanalytical methods. Cross-validation confirmed the interchangeability of the LC-MS/MS method developed in this study. ConclusionThis study developed and validated novel methods for CSA determination in whole blood and VAMS using LC-MS/MS. Clinical validation and proficiency testing confirmed their utility in routine clinical practice.

Long-term longitudinal monitoring of SARS CoV-2 in urban rivers and sewers of Nepal

In regions without adequate centralized wastewater treatment plants, sample collection from rivers and sewers can be an alternative sampling strategy for wastewater surveillance. This study aimed to assess the feasibility of alternative sampling strategies by testing samples collected from rivers (n = 246) and sewers (n = 244) in the Kathmandu Valley between March 2021 and February 2022. All samples were concentrated using the skimmed-milk flocculation method and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA was quantified using the nucleocapsid (N) and envelope (E) genes qPCR assays. Of the total, 75 % (371/490) of the samples tested positive using at least one qPCR assay, with concentrations ranging from 3.0 to 8.3 log10 gene copies/L. No significant correlation between concentrations of SARS-CoV-2 from both sewers and river with the number of confirmed coronavirus disease 2019 (COVID-19) cases in the Kathmandu valley was observed (p > 0.05). Despite the high concentration of SARS-CoV-2 in rivers and sewers, we hypothesize this finding to be a result of inaccurate number of clinical cases possibly due to inadequate clinical testing. This longitudinal study further supports the statement to consider sampling strategies from sewers and rivers for WBS in Nepal and other low and middle-income countries.

Preanalytical, Analytical and Postanalytical Analyses on Corynebacterium spp. and Actinomycetaceae in Urine Samples of Patients with Suspected Urinary Tract Infection-A Hypothesis-Forming Observational Study.

A hypothesis-forming exploratory cross-sectional assessment was conducted to assess the occurrence and relevance of Gram-positive rod-shaped bacteria like Corynebacterium spp. and Actinomycetaceae in human urine samples. In total, 1170 urine samples from 1031 inpatients with suspected urinary tract infection were assessed for culture-based growth of Gram-positive rod-shaped bacteria applying API Coryne assays, matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and in-house 16S rRNA gene sequencing. Overall, 502 different bacterial colonies from 346 urine samples taken from 324 inpatients were observed. The three quantitatively most abundant genera or genus clusters were Corynebacterium (254 isolates, 62%), Actinomyces/Winkia (79 isolates, 19%), and Actinotignum/Actinobaculum (29 isolates, 7%). Compared to sequencing, the diagnostic accuracy of all assessed competitor assays from the diagnostic routine was <80% for differentiation on the genus level and <30% for differentiation on the species level. Prolongated incubation for 4 days compared to 2 days resulted in additional detection of 15% of the totally recorded Gram-positive rod-shaped bacteria. An approximately 5-fold increased detection rate in mid-stream urine compared to urine acquired applying alternative sampling strategies was observed. In conclusion, in the rare event of the suspected clinical relevance of such findings, confirmatory testing with invasively sampled urine should be considered due to the high contamination rate observed in mid-stream urine. Confirmatory testing by DNA-sequencing methods should be considered if an exact identification of genus or species is regarded as relevant for the individual choice of the therapeutic strategy.

Pharmacokinetics-Based Pediatric Dose Evaluation and Optimization Using Saliva - A Case Study.

Understanding pharmacokinetics (PK) in children is a prerequisite to determine optimal pediatric dosing. As plasma sampling in children is challenging, alternative PK sampling strategies are needed. In this case study we evaluated the suitability of saliva as alternative PK matrix to simplify studies in infants, investigating metamizole, an analgesic used off-label in infants. Six plasma and 6 saliva PK sample collections were scheduled after a single intravenous dose of 10 mg/kg metamizole. Plasma/saliva pharmacometric (PMX) modeling of the active metabolites 4-methylaminoantipyrine (4-MAA) and 4-aminoantipyrine (4-AA) was performed. Various reduced plasma sampling scenarios were evaluated by PMX simulations. Saliva and plasma samples from 25 children were included (age range, 5-70 months; weight range, 8.7-24.8 kg). Distribution of metamizole metabolites between plasma and saliva was without delay. Estimated mean (individual range) saliva/plasma fractions of 4-MAA and 4-AA were 0.32 (0.05-0.57) and 0.57 (0.25-0.70), respectively. Residual variability of 4-MAA (4-AA) in saliva was 47% (28%) versus 17% (11%) in plasma. A simplified sampling scenario with up to 6 saliva samples combined with 1 plasma sample was associated with similar PK parameter estimates as the full plasma sampling scenario. This case study with metamizole shows increased PK variability in saliva compared to plasma, compromising its suitability as single matrix for PK studies in infants. Nonetheless, rich saliva sampling can reduce the number of plasma samples required for PK characterization, thereby facilitating the conduct of PK studies to optimize dosing in pediatric patients.

Simultaneous monitoring of seven antiepileptic drugs by dried blood spot and dried plasma spot sampling: method validation and clinical application of a LC–MS/MS-based technique

Alternative blood sampling strategy can enhance the application of therapeutic drug monitoring (TDM), then improve precision therapy and medication compliance. In developing nations, alternative sampling strategy that allows self-sampling and room temperature transport is especially important. This study validates the use of dried blood spot (DBS) and dried plasma spot (DPS) sampling along with liquid chromatography–tandem mass spectrometry (LC–MS/MS) for analyzing seven common antiepileptic drugs (AEDs) (phenytoin, lamotrigine, levetiracetam, topiramate, carbamazepine, oxcarbazepine and its active metabolite 10,11-dihydro-10-hydroxy carbamazepine) and evaluates their applicability to clinical practice. Following simple protein precipitation with acetonitrile, the AEDs were separated on a C18 column by gradient elution with a mobile phase consisting of acetonitrile–water–0.1% formic acid at a flow rate of 0.65 mL/min. The method provided linear analysis over the tested concentration ranges, with a total run time of 7 min. Intra- and inter-assay precision for all quality controls were ≤12% with accuracies of 85.9%–113%. The average extraction efficiencies were 69.0%–92.4% for DBS and 65.9%–96.5% for DPS, and no significant matrix effects were observed. The AEDs were stable in all samples for seven days at room temprature and 40°C. There was good correlation between the dry and wet plasma concentrations with greater accuracy for DPS compared to DBS indicating that alternative sampling strategy using DBS and DPS are suitable for monitoring the concentrations of AEDs with satisfied performance and logistical advantages.

Quantitative Analysis of Paleomagnetic Sampling Strategies

AbstractSampling strategies used in paleomagnetic studies play a crucial role in dictating the accuracy of our estimates of properties of the ancient geomagnetic field. However, there has been little quantitative analysis of optimal paleomagnetic sampling strategies and the community has instead defaulted to traditional practices that vary between laboratories. In this paper, we quantitatively evaluate the accuracy of alternative paleomagnetic sampling strategies through numerical experiments and an associated analytical framework. Our findings demonstrate a strong correspondence between the accuracy of an estimated paleopole position and the number of sites or independent readings of the time‐varying paleomagnetic field, whereas larger numbers of in‐site samples have a dwindling effect. This remains true even when a large proportion of the sample directions are spurious. This approach can be readily achieved in sedimentary sequences by distributing samples stratigraphically, considering each sample as an individual site. However, where the number of potential independent sites is inherently limited the collection of additional in‐site samples can improve the accuracy of the paleopole estimate (although with diminishing returns with increasing samples per site). Where an estimate of the magnitude of paleosecular variation is sought, multiple in‐site samples should be taken, but the optimal number is dependent on the expected fraction of outliers. The use of filters based on angular distance helps the accuracy of paleopole estimation, but leads to inaccurate estimates of paleosecular variation. We provide both analytical formulas and a series of interactive Jupyter notebooks allowing optimal sampling strategies to be developed from user‐informed expectations.

Efficient dynamic sampling of batch processes through operation recipes

The complexity of dynamic phenomena present in chemical processes often results in high evaluation costs of accurate first-principles models. This limits their real-time applicability, e.g. for advanced process control. A common solution is the derivation of simpler but faster data-driven surrogate models trained on simulated time series generated from dynamic samplings of a mechanistic model. For batch processes, known non-adaptive dynamic sampling methods lead to unrealistic or even infeasible operation cycles, raising the cost of generating simulated datasets with sufficient information content to train accurate surrogate models. An alternative sampling strategy is developed and analyzed, where sampled input trajectories are constrained to process knowledge in the form of parametrized operation recipes. The proposed methodology is tested for the case studies of full batch cycles of a crystallizer and a batch distillation column, showing that it is more efficient in terms of convergent simulations compared to an established dynamic sampling strategy.

The performance of model-based indices given alternative sampling strategies in a climate-adaptive survey design

Species-distribution shifts are becoming commonplace due to climate-driven change. Difficult decisions to modify survey extent and frequency are often made due to this change and constraining survey budgets. This often leads to spatially and temporally unbalanced survey coverage. Spatio-temporal models are increasingly used to account for spatially unbalanced sampling data when estimating abundance indices used for stock assessment, but their performance in these contexts has received little research attention. We therefore seek to answer two questions: (1) how well can a spatio-temporal model estimate the proportion of abundance in a new “climate-adaptive” spatial stratum? and (2) when sampling must be reduced, does annual sampling at reduced density or biennial sampling result in better model-based abundance indices? We develop a spatially varying coefficient model in the R package VAST using the eastern Bering Sea (EBS) bottom trawl survey and its northern Bering Sea (NBS) extension to address these questions. We first reduce the spatial extent of survey data for 30 out of 38 years of a real survey in the EBS and fit a spatio-temporal model to four commercially important species using these “data-reduction” scenarios. This shows that a spatio-temporal model generally produces similar trends and density estimates over time when large portions of the sampling domain are not sampled. However, when the central distribution of a population is not sampled the estimates are inaccurate and have higher uncertainty. We also conducted a simulation experiment conditioned upon estimates for walleye pollock (Gadus chalcogrammus) in the EBS and NBS. Many species in this region are experiencing distributional shifts attributable to climate change with species historically centered in the southeastern portion of the survey being increasingly encountered in the NBS. The NBS was occasionally surveyed in the past, but has been surveyed more regularly in recent years to document distributional shifts. Expanding the survey to the NBS is costly and given limited resources the utility of reducing survey frequency versus reducing sampling density to increase survey spatial extent is under debate. To address this question, we simulate survey data from alternative sampling designs that involve (1) annual full sampling, (2) reduced sampling in the NBS every year, or (3) biennial and full sampling in the NBS. Our results show that annual sampling, even with reduced sampling density, provides less biased abundance information than biennial sampling. We therefore conclude that ideally fishery-independent surveys should be conducted annually and spatio-temporal models can help to provide reliable estimates.

Evaluation of dried blood spots as an alternative sampling strategy for 5-fluorouracil monitoring: From method development to clinical application

Therapeutic drug monitoring (TDM) of 5-Fluorouracil (5-FU) is strongly recommended because of its large inter-individual pharmacokinetic variability, narrow therapeutic window, and incidence of toxicity. However, there are several factors that limit the application of TDM in clinical settings. Considering the intrinsic advantages of dried microsamples, such as minimally invasive sampling, analyte stability, and cost-effective logistics, this study aimed to develop a method for the determination of 5-FU in dried blood spots (DBS) using ultra-high liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and to evaluate its clinical application. Sample preparation was based on an aqueous extraction followed by protein precipitation. Separation was performed in an Acquity UPLC® HSS C18 (150 ×2.1 mm, 1.8 µm), and the mobile phases were water and acetonitrile with 0.5% acetic acid. The total run time was 5.5 min. The method was linear from 100 to 2000 ng/mL, precise (maximum CV% of 7.5%), and accurate (98.3–115.4%). The average recovery was 70%. Blood hematocrit had a minimal impact on the assay. DBS samples were stable for 21 days at 4, 25, and 45 °C. A total of 40 paired samples of plasma, capillary DBS, and venous DBS were analyzed. Median 5-FU concentrations were 444.7, 637.0, and 499.7 ng/mL for plasma, capillary DBS, and venous DBS, respectively. Capillary and plasma concentrations were significantly correlated (r > 0.90), but there was a lack of agreement between the methods, as capillary DBS levels were on average 146% of plasma. Venous DBS corresponded to 110% of the measured plasma concentrations, with a strong correlation (r > 0.97) and agreement between the methods. Our study is the first to report the use of DBS samples to quantify 5-FU. Further studies are needed to establish whether capillary samples can replace plasma.

Utilization of a population pharmacokinetic model to improve a busulfan test-dose strategy in allogeneic hematopoietic cell transplant recipients.

Busulfan is an alkylating agent used as part of conditioning chemotherapy regimens prior to allogeneic hematopoietic cell transplant (allo-HCT). Pharmacokinetic (PK)-guided test-dose strategies have been shown to improve the number of patients achieving busulfan exposure goals and improve clinical outcomes. However, current practices require extensive PK sampling. In this study, PK data were retrospectively collected from busulfan drug monitoring records from adult allo-HCT recipients who received once-daily intravenous busulfan at University of North Carolina Medical Center (UNCMC). A population pharmacokinetic (popPK) model was developed to identify sources of interindividual variability and evaluate alternative PK sampling strategies. A two-compartment model with covariate effects of actual body weight and sex best described the data. The typical value of clearance for an 83 kg male was estimated to be 11.21 L/hr. 59% of allo-HCT recipients were estimated to have met the UNCMC institutional MAC-exposure goal based on model post hoc estimates of clearance using all PK samples obtained following MAC-dosing. 57% of patients were estimated to have met this goal based on post hoc estimates using a single PK sample. Our results indicate once-daily, intravenous busulfan PK in adult allo-HCT recipients receiving MAC-dosing can be reasonably described by a popPK model and use of a sparse sampling strategy may be feasible for determining target exposure attainment following MAC-dosing. Use of a popPK model and sparse PK sampling strategy to carry out busulfan test-dose procedures could reduce healthcare costs and inconvenience to patients. This article is protected by copyright. All rights reserved.

The AERosol and TRACe gas Collector (AERTRACC): an online-measurement-controlled sampler for source-resolved emission analysis

Abstract. Probing sources of atmospheric pollution in complex environments often leads to the measurement and sampling of a mixture of different aerosol types due to fluctuations of the emissions or the atmospheric transport situation. Here, we present the AERosol and TRACe gas Collector (AERTRACC), a system for sampling various aerosol types independently on separate sampling media, controlled by parallel online measurements of particle, trace gas, and meteorological variables, like particle number or mass concentration, particle composition, trace gas concentration, and wind direction and speed. AERTRACC is incorporated into our mobile laboratory (MoLa) which houses online instruments that measure various physical and chemical aerosol properties, as well as trace gas concentrations. Based on preparatory online measurements with the whole MoLa setup, suitable parameters measured by these instruments are used to define individual sampling conditions for each targeted aerosol type using a dedicated software interface. Through evaluation of continuously online-measured data with regard to the sampling conditions, the sampler automatically switches between sampling and non-sampling for each of up to four samples, which can be collected in parallel. The particle phase and gas phase of each aerosol type, e.g., source emissions and background, are sampled onto separate filters with PM1 and PM10 cutoffs and thermal desorption tubes, respectively. Information on chemical compounds in the sampled aerosol is obtained by means of thermal desorption chemical ionization mass spectrometry (TD-CIMS) as the analysis method. The design, operation, and characterization of the sampler are presented. For in-field validation, wood-fired pizza oven emissions were sampled as targeted emissions separately from ambient background. Results show that the combination of well-chosen sampling conditions allows more efficient and effective separation of source-related aerosols from the background, as seen by the increases of particle number and mass concentration and concentration of organic aerosol types, with minimized loss of sampling time compared to alternative sampling strategies.

Impact and mitigation of sampling bias to determine viral spread: Evaluating discrete phylogeography through CTMC modeling and structured coalescent model approximations.

Bayesian phylogeographic inference is a powerful tool in molecular epidemiological studies, which enables reconstruction of the origin and subsequent geographic spread of pathogens. Such inference is, however, potentially affected by geographic sampling bias. Here, we investigated the impact of sampling bias on the spatiotemporal reconstruction of viral epidemics using Bayesian discrete phylogeographic models and explored different operational strategies to mitigate this impact. We considered the continuous-time Markov chain (CTMC) model and two structured coalescent approximations (Bayesian structured coalescent approximation [BASTA] and marginal approximation of the structured coalescent [MASCOT]). For each approach, we compared the estimated and simulated spatiotemporal histories in biased and unbiased conditions based on the simulated epidemics of rabies virus (RABV) in dogs in Morocco. While the reconstructed spatiotemporal histories were impacted by sampling bias for the three approaches, BASTA and MASCOT reconstructions were also biased when employing unbiased samples. Increasing the number of analyzed genomes led to more robust estimates at low sampling bias for the CTMC model. Alternative sampling strategies that maximize the spatiotemporal coverage greatly improved the inference at intermediate sampling bias for the CTMC model, and to a lesser extent, for BASTA and MASCOT. In contrast, allowing for time-varying population sizes in MASCOT resulted in robust inference. We further applied these approaches to two empirical datasets: a RABV dataset from the Philippines and a SARS-CoV-2 dataset describing its early spread across the world. In conclusion, sampling biases are ubiquitous in phylogeographic analyses but may be accommodated by increasing the sample size, balancing spatial and temporal composition in the samples, and informing structured coalescent models with reliable case count data.

The Impact of Supply Chain Integration on Operational Performance: An Empirical Study

Manufacturing companies nowadays are under constant pressure to deliver high-quality products at the lowest possible prices within the shortest possible time even under the most unpredictable economic situations. Supply chain integration has a critical impact on operational performance. Nevertheless, this impact has not been consistent and showed mixed results throughout the literature. This study aimed to examine the impact of technology management in terms of supply chain integration on operational performance. The research model was empirically validated using 317 valid survey responses from the Jordanian food and beverage industry, which were subjected to quantitative research design and regression analysis. Results showed that supply chain integration had a direct significant impact on operational performance, and all three dimensions of the theoretical model contributed significantly to operational performance. This study suggests the critical need to create and implement proper supply chain integration strategies and technologies, both internally and externally, to enhance their performance and competitive advantage. Moreover, future research needs to extend this work to other industries, cultures, and nations, while investigating the moderating or mediating effects of other key variables along with using alternative sampling strategies.

Cannabis practices among a gender-diverse sample of young adults

BackgroundGender is an important factor in understanding cannabis patterns, yet few studies have explored cannabis patterns among gender minority (GM) individuals – particularly among high-risk age groups including young adults. The evolving cannabis market is reshaping typical patterns of cannabis use in the U.S. The combination of these factors warrants increased efforts to examine cannabis practices in gender-diverse samples. MethodsOnline survey participants between 18 and 34 (N = 2377) from the U.S. provided information on cannabis practices from May – July 2021. Gender differences across several cannabis outcomes (onset, methods of consumption, product potency, frequency, and quantity) were assessed. Bivariate tests and multiple regression models examined associations between gender (cisgender men: n = 1020; cisgender women: n = 1178; and GM: n = 179) and cannabis outcomes adjusting for sociodemographic characteristics. ResultsIn regression models adjusted for sociodemographic characteristics, GM identity was associated with later age of onset and lower likelihood of daily use compared to cisgender men and women. Identifying as a GM person or cisgender woman was associated with fewer lifetime methods of consumption and less potent plant and concentrate product usage. ConclusionsFindings provide initial insights into potential gender differences in cannabis practices from a sample of heavy cannabis users. GM young adults report use patterns indicative of lower risk compared to cisgender men and women in our sample. Future investigations of gender differences in cannabis use that explore specific gender minority categories and that include alternative sampling strategies are needed to better understand differential risks associated with gender.

Self-collected gargle fluids and nasopharyngeal swabs as a strategy for molecular diagnostics of respiratory viruses

Diagnosis of respiratory viruses traditionally relies on deep oropharynx or nasopharynx swabs collected by healthcare workers (HCW). However, outpatients must make an appointment, and the procedure can cause discomfort in patients. Self-collecting has the potential as a strategy to improve participants’ willingness to participate in diagnostics, surveillance, or studies.We compared self-collected gargle fluids and nasopharyngeal swabs as a strategy for molecular diagnostics of respiratory viruses and compared the average cycle threshold (Ct)-values with those of samples collected by HCW. The study was conducted among technicians of the Laboratory of Clinical Microbiology and Infectious Diseases, Zwolle, the Netherlands, and their family members, between April 2019 and March 2020. It included a questionnaire regarding the severity and date of first symptoms and an assessment of the sampling experience. The primary outcome was the mean Ct of positive PCRs. Similar mean Ct values were obtained using self- or HCW-collected swabs. In addition, gargle fluids and self-swabbed specimens had comparable detection rates of respiratory viruses. Notably, most participants preferred gargling over self-swabbing. Interestingly, but not surprisingly, the time between the onset of symptoms and sampling was shorter in PCR-positive compared to PCR-negative participants.Though this study was abrogated by the SARS-CoV-2 pandemic, the results indicate that both self-swabs and gargle fluids are acceptable for diagnosing common respiratory viruses in the outpatient population, including influenza virus, rhinovirus, adenovirus, SARS-CoV-2 and endemic human coronaviruses. Gargling could be considered an alternative sampling strategy and may enhance willingness to participate in screenings or diagnostics for respiratory viruses.

Advancing Theory with Rigorous Problematizing Reviews: Giving-up Labels-Centric Literature Samples

Problematizing reviews seem doomed to be suspected of a lack of academic rigor because their advocates remain reluctant to use systematic sampling methodologies. Their reluctance is based on the premise that systematic reviews only cover pieces of research citing the name(s) of the construct(s) commonly used to study a phenomenon, what prevents any problematization of such construct(s). To envisage how to systematically sample existing literature without falling into such a sampling trap, we introduce a three-dimensional definition of constructs (label, intension, extension). Thanks to this framework, we first show that extension-centric samples outweigh the pitfalls that advocates of problematizing reviews associate with systematic methodologies relying on labels-centric samples. Second, we explain that this alternative sampling strategy enables reviews authors to excavate how the empirical objects being part of the extension of the phenomenon focal to the review are categorized by the theoretical schools that study them. Three ideal-typical of such categorizations emerge from our analysis—integrated, in halos or in Russian dolls—and we show that all of them are venues for problematization. This article contributes to the discussion of how to advance theory with review articles by theoretically developing a systematic sampling methodology enabling rigorous problematizing reviews.

Genetic individual identification from dried urine spots: A complementary tool to drug monitoring and anti‐doping testing

The collection of liquid biological matrices onto paper cards (dried matrix spots [DMS]) is becoming an alternative sampling strategy. The stability over time of molecules of interest for therapeutic, sport drug monitoring, and forensic toxicology on DMS has been recently investigated representing a reliable alternative to conventional analytical techniques. When a tampering of a urine sample in drug monitoring or doping control cases is suspected, it could be relevant to know whether genetic profiles useful for individual identification could be generated from urine samples spotted onto paper (dried urine spot [DUS]). To understand the influence of sex, storage conditions, and time on the quality and quantity of the DNA, five female and ten male urine samples were dispensed onto Whatman 903 paper and sampled after different storage conditions over time, from 1 to 12 weeks. Direct PCR was performed starting from 2‐mm punches collected from each spot amplifying a panel of markers useful for individual identification. The female DUS stored in different conditions produced genetic profiles fully matching the reference samples. The same result was obtained for the male DUS but using urine 30X concentrated by centrifugation instead of the original samples. Our data show that this approach is valid for genetic individual identification of urine samples spotted onto paper cards up to 12 weeks after deposition and could be easily incorporated in anti‐doping or drug screening protocols to help on the suspicion of evidence tampering or to solve questions on the reliability of samples collection.

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.

Sampling forests with terrestrial laser scanning.

Background and AimsTerrestrial laser scanners (TLSs) have successfully captured various properties of individual trees and have potential to further increase the quality and efficiency of forest surveys. However, TLSs are limited to line of sight observations, and forests are complex structural environments that can occlude TLS beams and thereby cause incomplete TLS samples. We evaluate the prevalence and sources of occlusion that limit line of sight to forest stems for TLS scans, assess the impacts of TLS sample incompleteness, and evaluate sampling strategies and data analysis techniques aimed at improving sample quality and representativeness.MethodsWe use a large number of TLS scans (761), taken across a 255 650-m2 area of forest with detailed field survey data: the Harvard Forest Global Earth Observatory (ForestGEO) (MA, USA). Sets of TLS returns are matched to stem positions in the field surveys to derive TLS-observed stem sets, which are compared with two additional stem sets derived solely from the field survey data: a set of stems within a fixed range from the TLS and a set of stems based on 2-D modelling of line of sight. Stem counts and densities are compared between the stem sets, and four alternative derivations of area to correct stem densities for the effects of occlusion are evaluated. Representation of diameter at breast height and species, drawn from the field survey data, are also compared between the stem sets.Key ResultsOcclusion from non-stem sources was the major influence on TLS line of sight. Transect and point TLS samples demonstrated better representativeness of some stem properties than did plots. Deriving sampled area from TLS scans improved estimates of stem density.ConclusionsTLS sampling efforts should consider alternative sampling strategies and move towards in-progress assessment of sample quality and dynamic adaptation of sampling.

Alternative Sampling Strategies in Therapeutic Drug Monitoring: Microsampling Growing Toward Maturity.

Alternative Sampling Strategies in Therapeutic Drug Monitoring: Microsampling Growing Toward Maturity.