Rapid quantification of Psilocybin with reversed-phase HPLC and single-wavelength detection

The alkaloid psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine) and the neurologically active psilocin (4-hydroxy-N,N-dimethyltryptamine) are the foremost compounds of pharmaceutical interest in Psilocybe mushrooms. As these compounds are infrequently analyzed in analytical labs, validated methods for rapid purity analysis are lacking. Newfound therapeutic use has invigorated academic and commercial interests in the molecules and new methods of production and available products are expanding. As a result, high-throughput methods of analysis for psilocybin must be improved to promptly determine chemical differences between mushroom genera or other sources of psilocybin and psilocin, as well as refined product purity. To address this, we developed an inexpensive HPLC technique for the efficient quantification of psilocybin and psilocin by using readily available equipment and dilute reagents. Aqueous ammonium formate (0.143 mM) was found to be preferable over techniques with much higher buffer concentrations or stronger acids for controlling psilocybin Zwitterion resolution. The chromatographic run time satisfied high-throughput analytical requirements with an efficient total runtime under 2 minutes. A standard octadecyl silica (C18) column provided excellent resolution between psilocybin and psilocin signals. The quality of the method was validated using certified analytical reference standards and was found to be accurate (3.5% bias, Psilocybin), reliable (0.32% RSD), and efficient (Psilocybin k’ = 1.78).

Invited Commentary: Standards, Inputs, and Outputs—Strategies for Improving Data-Sharing and Consortia-Based Epidemiologic Research

Data-sharing improves epidemiologic research, but the sharing of data frustrates epidemiologic researchers. The inefficiencies of current methods and options for data-sharing are increasingly documented and easily understood by any study group that has shared its data and any researcher who has received shared data. In this issue of the Journal, Temprosa et al. (Am J Epidemiol. XXX(XX):XXX–XXX) describe how the Consortium of Metabolomics Studies (COMETS) developed and deployed a flexible analytical platform to eliminate key pain points in large-scale metabolomics research. COMETS Analytics includes an online tool, but its cloud computing and technology are the supporting rather than the leading actors in this script. The COMETS team identified the need to standardize diverse and inconsistent metabolomics and covariate data and models across its many participating cohort studies, and then developed a flexible tool that gave its member studies choices about how they wanted to meet the consortium’s analytical requirements. Different specialties will have different specific research needs and will probably continue to use and develop an array of diverse analytical and technical solutions for their projects. COMETS Analytics shows how important—and enabling—the upstream attention to data standards and data consistency is to producing high-quality metabolomics, consortia-based, and large-scale epidemiology research.

Validation of Multispectral Imaging (MSI) technology for food and feed analysis

The current testing environment for food and feed samples is complex and uses a wide variety of analytical technologies that range from chemical to PCR-based. Whilst these approaches can be deemed fit for purpose in terms of their final analytical result, they tend to be highly specialised and require considerable frontend processing to ensure that the target analyte can be reliably detected and quantified. These combined sample processing and analytical requirements typically impact on testing times and have associated cost implications that must be factored into routine testing and monitoring applications. MSI uses multiple discrete and informative wavelengths covering regions such as the UV and near infra-red spectrum to quickly determine surface colour, texture and possible chemical composition. Compared to traditional molecular biology approaches utilising DNA extraction followed by PCR-based analyses, MSI can simplify and reduce the time/costs associated with sample analysis. It is rapid and non-destructive.

Groundwater resources monitoring during unconventional oil and gas extraction: South African laboratory analytical capabilities

Groundwater resource quality monitoring before, during, and after unconventional oil and gas (UOG) extraction would assist in protecting groundwater resources. Limited laboratory analytical capacity may, however, hamper effective monitoring. We assessed South African (SA) laboratory analytical capabilities for specific groundwater monitoring parameters relevant to UOG extraction. We found a limited capacity to analyse for most of the UOG extraction–related groundwater monitoring parameters and that most of the surveyed laboratories are not planning to increase their analyses capacity to cater for UOG extraction. This issue must be addressed urgently if SA wants to proceed with UOG extraction. Policy recommendations include that South Africa should develop a specialised UOG extraction monitoring laboratory to cater for analytical needs. Such capacity could also address the analytical requirements for the rest of the African region during UOG extraction.

Null and Void? Errors in Meta-analysis on Perceptual Disfluency and Recommendations to Improve Meta-analytical Reproducibility

In the 2018 meta-analysis of Educational Psychology Review entitled “Null effects of perceptual disfluency on learning outcomes in a text-based educational context” by Xie, Zhou, and Liu, we identify some errors and inconsistencies in both the methodological approach and the reported results regarding coding and effect sizes. While from a technical point of view the meta-analysis aligns with current meta-analytical guidelines (e.g., PRISMA) and conforms to general meta-analytical requirements (e.g., considering publication bias), it exemplifies certain insufficient practices in the creation and review of meta-analysis. We criticize the lack of transparency and negligence of open-science practices in the generation and reporting of results, which complicate evaluation of the meta-analytical reproducibility, especially given the flexibility in subjective choices regarding the analytical approach and the flexibility in creating the database. Here we present a framework applicable to pre- and post-publication review on improving the Methods Reproducibility of meta-analysis. Based on considerations of the transparency and openness (TOP)-guidlines (Nosek et al. Science 348: 1422–1425, 2015), the Reproducibility Enhancement Principles (REP; Stodden et al. Science 354:1240–1241, 2016), and recommendations by Lakens et al. (BMC Psychology 4: Article 24, 2016), we outline Computational Reproducibility (Level 1), Computational Verification (Level 2), Analysis Reproducibility (Level 3), and Outcome Reproducibility (Level 4). Applying reproducibility checks to TRANSFER performance as the chosen outcome variable, we found Xie’s and colleagues’ results to be (rather) robust. Yet, regarding RECALL performance and the moderator analysis, the identified problems raise doubts about the credibility of the reported results.

How to meet ISO15189:2012 pre-analytical requirements in clinical laboratories? A consensus document by the EFLM WG-PRE

The International Organization for Standardization (ISO) 15189:2012 standard aims to improve quality in medical laboratories through standardization of all key elements in the total testing process, including the pre-analytical phase. It is hence essential that accreditation bodies, assessing laboratories against ISO15189:2012, pay sufficient attention to auditing pre-analytical activities. However, there are significant differences in how technical auditors interpret the pre-analytical requirements described in ISO15189:2012. In this consensus document, the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for Pre-analytical Phase (WG-PRE) sets out to review pre-analytical requirements contained in ISO15189:2012 and provide guidance for laboratories on how to meet these requirements. The target audience for this consensus document is laboratory professionals who wish to improve the quality of the pre-analytical phase in their laboratory. For each of the ISO requirements described in ISO15189:2012, members of EFLM WG-PRE agreed by consensus on minimal recommendations and best-in-class solutions. The minimal consensus recommendation was defined as the minimal specification which laboratories should implement in their quality management system to adequately address the pre-analytical requirement described in ISO15189:2012. The best-in-class solution describes the current state-of-the-art in fulfilling a particular pre-analytical requirement in ISO15189:2012. We fully acknowledge that not every laboratory has the means to implement these best-in-class solutions, but we hope to challenge laboratories in critically evaluating and improving their current procedures by providing this expanded guidance.

Predictive Analytics for the KMAP-O Model in Design and Evaluation of Diabetes Care Management Research

This is a commentary on methodological challenges and analytical requirements in designing an evaluation of the knowledge, motivation, attitude, preventive practice-outcome (KMAP-O) model for selfcare management of diabetes. Critical issues pertaining to an investigation of the dose-response relationship between the intervention program and outcomes, the comparative effectiveness evaluation, and the lengths of observation were noted. Although numerous publications on factors influencing diabetes care and control were systematically reviewed and documented in the literature, scientific results on artificial intelligence research remain to be uncovered. To optimizing the knowledge and clinical practice in selfcare management, specific methodological approaches to predictive analytics are suggested for future clinical studies, using a comprehensive behavioral system such as the KMAP-O model.

Survey of patient perception of pre-analytical requirements for blood testing in the UK and RoI

Background A patient survey developed by the Pre-Analytical Phase Special Interest Group of the Association for Clinical Biochemistry and Laboratory Medicine (ACB-PA-SIG) was conducted during November and December 2019. The survey aimed to determine the quality of information provided to patients in preparation for their blood test(s). In addition, the ACB-PA-SIG provide a number of recommendations, which, if adopted, may yield higher quality test results and improve patient management. Methods The survey was distributed at phlebotomy suites in two Hospitals: Ipswich Hospital (United Kingdom [UK]), and Cork University Hospital (Republic of Ireland [RoI]). Results Overall, 235 survey responses were received from the two sites. A total of 103 respondents received no information about preparing for their blood test and 92 had been told they did not need to fast. None of the patients surveyed had been instructed to fast for 12 h. Twenty-two patients had been told to avoid certain foods, drinks or medication, 14 were told to avoid strenuous activity and 41 respondents had been informed of the need to avoid alcohol/smoking prior to their blood test. Overall, only approximately 78 felt well informed about the blood taking process. Conclusions Based on the results of this survey, the ACB-PA-SIG conclude that: (1) clinicians should provide clear written information to patients regarding pre-analytical requirements; and (2) effective communication between laboratories and General Practitioners is required to disseminate information. In this paper, the ACB-PA-SIG provide a list of pre-analytical recommendations to standardize and improve practice across the UK and RoI.

Determination of icariin content in dietary supplements by high performance liquid chromatography

High performance liquid chromatography with PDA detector technique has been applied to analyze the Icariin content in dietary supplements that contains the ingredient of Epimedium. Sample preparation used methanol solvent and ultrasonic technique. Analytical procedure performed on HPLC Alliance-Waters e2695 system with chromatographic column C18 Reliant 2 (250 mm × 4.6 mm, 5 µm) and gradient between acetonitril and phosphoric acid 1% solution in 20 minutes. The validation results showed that the method had good specificity, linear calibration curve in the range of 0.42 - 42 µg/mL, the repeatability and recovery of the method meet the analytical requirements according to AOAC. The method has been deployed to analyze some dietary supplement products on the market and give Icariin content 6.15 - 54.3 mg/g (in dietary supplement) and 6.47 - 24.5 g/100g (in powder extracts).

Automated Measurement of Plasma Cell-Free Hemoglobin Using the Hemolysis Index Check Function

Background The Roche Cobas chemistry analyzer’s hemolysis index (HI) check function can directly report hemoglobin (Hb) concentrations. We aimed to validate the HI check function for the measurement of plasma cell-free Hb. Methods Plasma samples (6 μl) were taken by the analyzer and diluted in normal saline to measure the absorbance for Hb at 570 and 600 nm. Hb concentrations were calculated based on the molar extinction coefficient. Imprecision, lower limit of quantification (LLOQ), and analytical measurement range (AMR) of the assay were evaluated. The accuracy was determined by comparing the results between the new method and an existing spectrophotometric method. We further studied interference of icterus and lipemia and carryover. The performance of the assay in proficiency testing was also evaluated. The reference range was transferred from the existing method. Results Within-run and total CVs were 1.7%–4.2% and 2.1%–7.0%, respectively (n = 20). The LLOQ was 11 mg/dL (CV = 8.1%) with the upper limit of AMR of 506 mg/dL. The results of the new method correlated well with the existing reference assay: Y (new method) = 0.974 x (reference method) + 4.9, r = 0.9990, n = 52. Bilirubin with a concentration up to 60 mg/dL and lipemic index up to 389 did not show significant interference. No significant carryover was detected. The average standard deviation index in proficiency testing was 0.03 ± 0.29. The reference range was <22 mg/dL. Conclusions Plasma cell-free Hb measurement using the HI check function meets the analytical requirements of the plasma cell-free Hb assays. It is simple and cost-effective.