Implementation of field detection devices for antimalarial quality screening in Lao PDR-A cost-effectiveness analysis.

Substandard and falsified (SF) antimalarials have devastating consequences including increased morbidity, mortality and economic losses. Portable medicine quality screening devices are increasingly available, but whether their use for the detection of SF antimalarials is cost-effective is not known. We evaluated the cost-effectiveness of introducing such devices in post-market surveillance in pharmacies in Laos, conservatively focusing on their outcome in detecting SF artemisinin-based combination therapies (ACTs). We simulated the deployment of six portable screening devices: two handheld near-infrared [MicroPHAZIR RX, NIR-S-G1], two handheld Raman [Progeny, TruScan RM]; one portable mid-infrared [4500a FTIR] spectrometers, and single-use disposable paper analytical devices [PADs]. We considered two scenarios with high and low levels of SF ACTs. Different sampling strategies in which medicine inspectors would test 1, 2, or 3 sample(s) of each brand of ACT were evaluated. Costs of inspection including device procurement, inspector time, reagents, reference testing, and replacement with genuine ACTs were estimated. Outcomes were measured as disability adjusted life years (DALYs) and incremental cost-effectiveness ratios were estimated for each device compared with a baseline of visual inspections alone. In the scenario with high levels of SF ACTs, all devices were cost-effective with a 1-sample strategy. In the scenario of low levels of SF ACTs, only four devices (MicroPHAZIR RX, 4500a FTIR, NIR-S-G1, and PADs) were cost-effective with a 1-sample strategy. In the multi-way comparative analysis, in both scenarios the NIR-S-G1 testing 2 samples was the most cost-effective option. Routine inspection of ACT quality using portable screening devices is likely to be cost-effective in the Laos context. This work should encourage policy-makers or regulators to further investigate investment in portable screening devices to detect SF medicines and reduce their associated undesired health and economic burdens.

Hydrotropic Solubilization in Pharmaceutical Analysis: Origin, Evolution, Cumulative Trend and Precise Applications.

The hydrotropy though existing as precisely used scientific approach assisting solubilization for all. It took 66 long years to use the concept in drug analysis since its inception in 1916 and first use for facilitation of drug solubility toward better pharmaceutical analysis in 1982. Considering the unending importance in pharmaceutical sciences and thereby in analysis, it's a necessity to comprehensively outlook the origin, evolution, cumulative trend and precise applications in pharmaceutical analysis. Achieved hereby with chronological and comparative assessment of the studies published pertaining to solubility enhancement of poorly soluble drugs with use of hydrotropic agents alone or in combination for assisting pharmaceutical analysis. The thorough literature searches resulted into 77 references over a span of about 38 years. This comprehensive review critically evaluates existing literature; to our surprise we found Ibuprofen sodium, Lignocaine, Niacinamide and Metformin HCl as atypical hydrotropic agents. We also compared herein mono and mixed approaches which indicated prevalence of mono - hydrotropy over mixed. The possible mechanisms behind solubilization are presented for an additional insight. An essential effort has been made to state arbitrary classification to assist in future applications. The obvious purpose of this study was to collectively evaluate the crucial role of hydrotropic agents in pharmaceutical analyses for better drug delivery. This comprehensive review covers all details since inception to the updates till date which will definitely act as appropriate guideline for pharmaceutical analyst's in need of hydrotropy to assist pharmaceutical analysis for therapies today and tomorrow.

A handmade DNA extraction kit using laundry powder; insights on simplicity, cost-efficiency, rapidity, safety and the quality of purified DNA.

Here we describe an in-house kit for high throughput DNA extraction using laundry detergent. A simplified lysis buffer made only from 0.08 M EDTA, 0.1 M Tris, and laundry powder is the core of our protocol. We extracted genomic DNA from 150 µL of whole blood collected from different farm animals and compared the performance to both the DNeasy Blood & Tissue Kit (Qiagen) and the widely used salting-out procedure. An evaluation of the concentration and quality of the extracted DNA was then assessed by the NanoDrop absorption spectra, agarose gel migration, amplification in PCR and the Sanger sequencing. The in-house kit successfully extracted clean DNA from all blood samples, and discernably outperformed the commercial kits and the original salting-out procedure in the sense of the simplicity, cost-efficiency, quantity, and the quality of purified DNA. Apart from replacing proteinase K and the sodium dodecyl sulfate treatment by the laundry detergent, our protocol instructs a lysis buffer that eliminates sucrose, Triton X-100, MgCl2, NH4Cl, and KCl. Our handmade kit might be of interest for laboratories in underdeveloped countries with a budget shortage or applications in difficult field conditions, for example, when fridge storage for proteinase K cannot be ensured.

Rapid Screening α-Glucosidase Inhibitors from Natural Products by At-Line Nanofractionation with Parallel Mass Spectrometry and Bioactivity Assessment.

In this study, a novel at-line nanofractionation screening platform was successfully developed for the rapid screening and identification of α-glucosidase inhibitors from natural products. A time-course bioassay based on high density well-plates was performed in parallel with high resolution mass spectrometry (MS), providing a straightforward and rapid procedure to simultaneously obtain chemical and biological information of active compounds. Through multiple nanofractionations into the same well-plate and comparisons of the orthogonal separation results of hydrophilic interaction liquid chromatography (HILIC) and reversed-phase liquid chromatography (RPLC), the α-glucosidase inhibitors can be accurately identified from co-eluates. The screening platform was comprehensively evaluated and validated, and was applied to the screenings of green tea polyphenols and Ginkgo folium flavonoids. After accurate peak shape and retention time matching between the bioactivity chromatograms and MS chromatograms, ten α-glucosidase inhibitors were successfully screened out and identified. The proposed screening method is rapid, effective and can avoid ignoring low abundant/active inhibitors.

Finding a reliable limit of detection in the NIR determination of residual moisture in a freeze-dried drug product.

The validation of an analytical method in the pharmaceutical industry follows strictly regulated guidelines. The introduction of multivariable calibration methods requires a revision of these recommendations, since some of them are contradictory regarding the limit of detection (LOD). This work compares the LOD values obtained using pseudounivariate and multivariate procedures in the PLS-NIR determination of residual moisture content (RMC) in a freeze-dried drug. As NIR has proved to be more precise than Karl-Fischer at low RMC values, LOD has been estimated by ordinary and by orthogonal least squares regression. The precision of the RMC determination in approx. 2000 industrial vials was used as an indirect evidence of the reliability of the LOD values obtained. The effect of reducing the number of calibration samples and increasing the RMC values have also been studied. No significant differences were observed using a number of calibration samples ≥ 20. Based on our findings, when the size of the calibration sample set is high and the range of RMC values is close to the limit, the LOD estimated with the ICH formula and using orthogonal regression should be recommended. If water content moves away, the ICH formula should be replaced by the LODOS equation as a practical, reliable and simple procedure.

Reference-mean-centered statistical quality control.

Background: Statistical quality control (SQC) procedures generally use rejection limits centered on the stable mean of the results obtained for a control material by the analyzing instrument. However, for instruments with significant bias, re-centering the limits on a different value could improve the control procedures from the viewpoint of patient safety. Methods: A statistical model was used to assess the effect of shifting the rejection limits of the control procedure relative to the instrument mean on the number of erroneous results reported as a result of an increase in the systematic error of the measurement procedure due to an out-of-control condition. The behaviors of control procedures of type 1ks (k = 2, 2.5, 3) were studied when applied to analytical processes with different capabilities (σ = 3, 4, 6). Results: For measuring instruments with bias, shifting the rejection limits in the direction opposite to the bias improves the ability of the quality control procedure to limit the risk posed to patients in a systematic out-of-control condition. The maximum benefit is obtained when the displacement is equal to the bias of the instrument, that is, when the rejection limits are centered on the reference mean of the control material. The strategy is sensitive to error in estimating the bias. Shifting the limits more than the instrument's bias disproportionately increases the risk to patients. This effect should be considered in SQC planning for systems running the same test on multiple instruments. Conclusions: Centering the control rule on the reference mean is a potentially useful strategy for SQC planning based on risk management for measuring instruments with significant and stable uncorrected bias. Low uncertainty in estimating bias is necessary for this approach not to be counterproductive.

Chemical Identification and Confirmation of Contact Allergens.

Identification of the etiological chemical agent(s) associated with a case(s) of allergic contact dermatitis (ACD) is important for both patient management and public health surveillance. Traditional patch testing can identify chemical allergens to which the patient is allergic. Confirmation of allergen presence in the causative ACD-associated material is presently dependent on labeling information, which may not list the allergenic chemical on the product label or safety data sheet. Dermatologists have expressed concern over the lack of laboratory support for chemical allergen identification and possibly quantification from patients' ACD-associated products. The aim of this review was to provide the clinician a primer to better understand the analytical chemistry of contact allergen confirmation and unknown identification, including types of analyses, required instrumentation, identification levels of confidence decision tree, limitations, and costs.

Assessment of fish communities using environmental DNA: Effect of spatial sampling design in lentic systems of different sizes.

Freshwater fish biodiversity is quickly decreasing and requires effective monitoring and conservation. Environmental DNA (eDNA)-based methods have been shown to be highly sensitive and cost-efficient for aquatic biodiversity surveys, but few studies have systematically investigated how spatial sampling design affects eDNA-detected fish communities across lentic systems of different sizes. We compared the spatial patterns of fish diversity determined using eDNA in three lakes of small (SL; 3 ha), medium (ML; 122 ha) and large (LL; 4,343 ha) size using a spatially explicit grid sampling method. A total of 100 water samples (including nine, 17 and 18 shoreline samples and six, 14 and 36 interior samples from SL, ML and LL, respectively) were collected, and fish communities were analysed using eDNA metabarcoding of the mitochondrial 12S region. Together, 30, 35 and 41 fish taxa were detected in samples from SL, ML, and LL, respectively. We observed that eDNA from shoreline samples effectively captured the majority of the fish diversity of entire waterbodies, and pooled samples recovered fewer species than individually processed samples. Significant spatial autocorrelations between fish communities within 250 m and 2 km of each other were detected in ML and LL, respectively. Additionally, the relative sequence abundances of many fish species exhibited spatial distribution patterns that correlated with their typical habitat occupation. Overall, our results support the validity of a shoreline sampling strategy for eDNA-based fish community surveys in lentic systems but also suggest that a spatially comprehensive sampling design can reveal finer distribution patterns of individual species.

Plasmon-coupled 3D porous hotspot architecture for super-sensitive quantitative SERS sensing of toxic substances on real sample surfaces.

This paper reports a facile, fast, and cost-effective method for the synthesis of three-dimensional (3D) porous AgNPs/Cu composites as SERS substrates for the super-sensitive and quantitative detection of food organic contaminations. Due to the 3D porous hotspot architecture and the strong plasmonic coupling between Ag and Cu, the porous AgNPs/Cu substrate achieves ultrasensitive detection of multiple analytes as low as 10-11 M (crystal violet, CV), 10-9 M (malachite green, MG), 10-11 M (acephate), and 10-9 M (thiram) even with a portable Raman device. Moreover, this 3D solid substrate has good signal uniformity (RSD < 11%) and superior stability (<14% signal loss), allowing for practical SERS detections. Importantly, by simply wiping the real sample surface using the substrate, it successfully detects CV and MG residues on crayfish, and the limit of detection (LOD) of CV and MG is determined to be 1.14 × 10-9 M and 0.94 × 10-7 M, respectively. Further, the substrate can also be applied to detect acephate on eggplant with a LOD of 1.41 × 10-9 M and thiram on an apple surface with a LOD of 1.04 × 10-7 M. Note that all these SERS detections on real samples have a broad dynamic concentration range and a good linear dependence. As a "proof of concept", multi-component detection on a real sample has also been demonstrated. This 3D solid substrate possesses excellent detection sensitivity, diversity, and accuracy, which allows rapid and reliable determination of toxic substances in foods.

Mycotoxin Testing Paradigm: Challenges and Opportunities for the Future.

Mycotoxins are one of the great global challenges to agri-food and feed safety. Industry requires fast, reliable, and economical testing methods for the most important regulated mycotoxins to manage this problem. Climate change and changes in agricultural practice are complicating this situation, triggering the movement of some mycotoxins into new regions, which are unprepared for their management. Modern LC-tandem MS (LC-MS/MS) instruments have addressed this analytical challenge, but such instruments are expensive and require highly qualified personnel and dedicated facilities. As a result of these limitations, traditional LC-MS/MS is not amenable for use on farms or at small to midsized processing facilities, such as a grain elevator. To address the need for on-site rapid testing, the mycotoxin community has focused on antibody-based and spectrophotometric approaches. The development of innovative technologies such as miniaturized MS would allow for the acquisition of more information on mixtures of toxins present in a sample at costs comparable to those of the existing rapid methods such as ELISA. The capital costs are higher, but it would reduce per-sample testing costs and time requirements and provide better value for money while maintaining the accuracy and selectivity achieved in a laboratory setting. In this article, we review the available techniques and contrast them in the context of three main criteria: method performance, speed of analysis, and cost. We define the integration of these three parameters as the "mycotoxin testing paradigm."

Doing more with less: Evaluation of the use of high linear velocities in preparative supercritical fluid chromatography.

The evaluation of higher than typical linear velocities is discussed for supercritical fluid chromatographic purifications on the preparative scale. SFC separation efficiency suffers far less at high linear velocities than HPLC by the rapid mass transfer of analytes carried by compressed CO2 through the stationary phase. The technique is discussed using chiral test compounds and columns. In many cases, running at high linear velocities can yield significant time savings and decreased consumption of mobile phase solvent, while also lowering energy consumption. Within the practical limitations of commercial instrumentation, using 20 µm particles can aid in achieving higher linear velocities not attainable with smaller 5 µm particles, particularly when running with high percentages of organic co-solvent. Use of larger particles for the stationary phase also lowers the associated column cost. These benefits can yield an overall purification process that is more productive and environmentally friendly.

High-performance thin-layer chromatography/bioautography and liquid chromatography-mass spectrometry hyphenated with chemometrics for the quality assessment of Morus alba samples.

Quality control is a crucial step in the production of effective and safe herbal remedies. The aim of this study was to develop a new, simple, and high throughput procedure for the quality assessment of herbal drugs using a high-performance thin-layer chromatography (HPTLC)/bioautography and UPLC-MS/MS approach combined with chemometrics. This was exemplarily shown for Morus alba L. root bark (sang bái pí; SBP). Bioautography assays were developed for the identification of constituents with radical scavenging (DPPH assay) and antimicrobial activities (Bacillus subtilis, Escherichia coli) of 18 different M. alba samples, which was supported by UPLC-MS/MS analysis. Further, the combination of bioautography and chemometrics identified those samples with the most bioactive constituents. Plant materials collected from Serbia (11 samples) showed higher both radical scavenging and antimicrobial activities compared to samples provided from China (7 samples). Principal component analysis (PCA) confirmed the discrimination of geographically different samples and recognized their main markers responsible for differences between Serbian and Chinese samples. Most importantly for quality assessment, the combined HPTLC/bioautography and UPLC-MS/MS approach not only allowed for a fast chemical profiling of the investigated samples and their unambiguous identification, but also for the disclosure of major and minor bioactive constituents present in SBP.

A magnetic hyperbranched polyamide amine-based quick, easy, cheap, effective, rugged and safe method for the detection of organophosphorus pesticide residues.

Magnetic hyperbranched polyamide amine (MHPA) was successfully prepared and applied as a QuEChERS adsorbent to the gas chromatograph-mass spectrometer (GC-MS) detection of organophosphorus pesticides (OPPs) in orange juice. Abundant in amino types and the structure of hyperbranched organic chains make MHPA to possess the clean-up function integrating classic clean-up agent primary secondary amine (PSA) with C18 modified silica, and the magnetic property makes the operation easier and more time-saving. Compared the performance with classical clean-up agents of PSA and C18, better results were obtained with MHPA as adsorbent for the detection of 11 OPPs. Recoveries ranged from 75.2 to 116.2% with the relative standard deviation (RSD) of 4.1-18.9% and the limit of detection (LOD) of 0.74-8.16 ng/g. Results showed that using MHPA as adsorbent for QuEChERS sample preparation is an effective alternative with simplicity and rapidity.

Advances in Optical Sensing and Bioanalysis Enabled by 3D Printing.

The recent explosion of 3D printing applications in scientific literature has expanded the speed and effectiveness of analytical technological development. 3D printing allows for manufacture that is simply designed in software and printed in-house with nearly no constraints on geometry, and analytical methodologies can thus be prototyped and optimized with little difficulty. The versatility of methods and materials available allows the analytical chemist or biologist to fine-tune both the structural and functional portions of their apparatus. This flexibility has more recently been extended to optical-based bioanalysis, with higher resolution techniques and new printing materials opening the door for a wider variety of optical components, plasmonic surfaces, optical interfaces, and biomimetic systems that can be made in the laboratory. There have been discussions and reviews of various aspects of 3D printing technologies in analytical chemistry; this Review highlights recent literature and trends in their applications to optical sensing and bioanalysis.

Incurred sample reproducibility: 10 years of experiences: views and recommendations from the European Bioanalysis Forum.

With 10 years of experiences on incurred sample reanalysis (ISR) as an integrated part of regulated bioanalysis, the European Bioanalysis Forum has reflected on the implementation and the use of ISR. Three surveys were conducted in 2016 and 2017 as a revisit of the ISR experiences within European pharmaceutical industry and contract research organizations: has ISR become a tool for postvalidation and process check of a bioanalytical method performance and has ISR become a routine in our laboratories? Do we agree on the interpretation of guidelines/guidance and are we aligned in our approach - among others?

In vitro assessment of hepatotoxicity by metabolomics: a review.

Omics technologies, and in particular metabolomics, have received an increasing attention during the assessment of hepatotoxicity in vitro. However, at present, a consensus on good metabolomics practices has yet to be reached. Therefore, in this review, a range of experimental approaches, applied methodologies, and data processing workflows are compared and critically evaluated. Experimental designs among the studies are similar, reporting the use of primary hepatocytes or hepatic cell lines as the most frequently used cell sources. Experiments are usually conducted in short time-frames (< 48 h) at sub-toxic dosages. Applied sample preparations are protein precipitation or Bligh-and-Dyer extraction. Most analytical platforms rely on chromatographic separations with mass spectrometric detection using high-resolution instruments. Untargeted metabolomics was typically used to allow the simultaneous detection of several classes of the metabolome, including endogenous metabolites that are not initially linked to toxicity. This non-biased detection platform is a valuable tool for generating hypothesis-based mechanistic research. The most frequently reported metabolites that are altered under toxicological impulses are alanine, lactate, and proline, which are often correlated. Other unspecific biomarkers of hepatotoxicity in vitro are the down-regulation of choline, glutathione, and 3-phospho-glycerate. Disruptions on the Krebs cycle are associated with increased glutamate, tryptophan, and valine. Phospholipid alterations are described in steatosis, lipo-apoptosis, and oxidative stress. Although there is a growing trend towards quality control, data analysis procedures do often not follow good contemporary metabolomics practices, which include feature filtering, false-discovery rate correction, and reporting the confidence of metabolite annotation. The currently annotated biomarkers can be used to identify hepatotoxicity in general and provide, to a certain extent, a tool for mechanistic distinction.

Risk-based approach for method development in pharmaceutical quality control context: A critical review.

Pharmaceutical regulatory bodies increasingly require the implementation of systematic approaches in pharmaceutical product development. Quality control methods play a key role in the control strategy of drugs manufacturing to assure their quality. A risk-based approach in the analytical method development is strongly recommended to ensure that the method performances fit the purpose of the method during its entire life-cycle. In the last decade, analytical quality by design (AQbD), as risk management oriented methodology, has been progressively integrated with method development for fulfilling this objective. This approach has successfully allowed the quality to be designed into the analytical processes by obtaining a deep understanding of the procedures. In this paper the AQbD workflow and its application in the development of methods to be used for pharmaceutical quality control have been treated and discussed. Recent publications regarding how AQbD has been applied in separation techniques were reviewed. The different development strategies have been also showcased, highlighting their advantages and disadvantages, in order to give a useful overview.