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.

Microfluidic Paper-Based Analytical Device for Histidine Determination.

A laminated paper-based analytical device (LPAD) for histidine detection was fabricated from a chromatography filtration paper and laminate films. Histidine recognition was effected by histidyl-tRNA synthetase (HisRS), and its detection was signaled colorimetrically based on the molybdenum blue reaction. The analytical conditions and detectable concentration range of histidine were examined. The method provided selective quantification from 1 to 100 µM histidine. LPAD fabrication is considerably simple, involving only the craft-cutting of the chromatography filtration paper and laminate film, and is cost-effective.

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.

Basal serum luteinising hormone cut-off, and its utility and cost-effectiveness for aiding the diagnosis of the onset of puberty in girls with early stages of breast development.

OBJECTIVE: To determine basal and gonadotrophin-releasing hormone analogue (GnRHa)-stimulated peak luteinising hormone (LH) cut-offs to diagnose onset of early or normal puberty in girls with each Tanner stage of breast (II and III). DESIGN, PATIENTS AND MEASUREMENTS: A retrospective study of 601 girls with breast onset before 8 years of age who underwent GnRHa test was conducted. Patients were categorized as CPP and premature thelarche. Each group was divided into two subgroups; Tanner II and III. Cost-effectiveness analysis was performed. RESULTS: In comparison with basal LH cut-off of 0.3 IU/L, basal LH cut-off of 0.2 IU/L had comparable specificity (Tanner II: 98.0% vs 94.8%, Tanner III: 98.8% vs 93.8%), but greater sensitivity (Tanner II: 28.3% vs 41.7%, Tanner III: 45.2% vs 59.3%). Specificity of basal LH cut-off of 0.2 IU/L was not inferior to that of the traditionally used peak LH of 5 IU/L. Using basal LH cut-off of 0.2 IU/L followed by GnRHa test in girls with negative basal LH was more cost-saving when compared with using the cut-off of 0.3 IU/L. Moreover, using basal LH cut-off of 0.2 IU/L followed by GnRHa test provided a cost reduction when compared with performing GnRHa test in all patients. CONCLUSIONS: Basal serum LH cut-off of 0.2 IU/L could be a simple and cost-saving tool for initial diagnosis of onset of early or normal puberty in girls with Tanner II and III before proceeding to GnRH testing.

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.

Microwave Assisted Synthesis of N-Doped Carbon Dots: an Easy, Fast and Cheap Sensor for Determination of Aspartic Acid in Sport Supplements.

In this work, determination of aspartic acid by N-doped carbon dots (N-CDs) was studied at optimum condition. Characterization and morphology of surface of N-CDs were carried out by FT-IR and HRTEM. N-doped carbon dots size was 10 nm. Quenching was very fast after addition of aspartic acid that is an important property of this sensor. Optimum conditions for pH and excitation wavelength were 8 and 360 nm, respectively. Linear dynamic range and limit of detection for aspartic acid were 0.5-50 µM and 90 nM, respectively. This method was used for aspartic acid determination in human serum and sport supplement powder as real samples. Performance of this sensor was also compared with other fluorescent sensors.

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.

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.

Study of Hybrid PVA/MA/TEOS Pervaporation Membrane and Evaluation of Energy Requirement for Desalination by Pervaporation.

Desalination by pervaporation is a membrane process that is yet to be realized for commercial application. To investigate the feasibility and viability of scaling up, a process engineering model was developed to evaluate the energy requirement based on the experimental study of a hybrid polyvinyl alcohol/maleic acid/tetraethyl orthosilicate (PVA/MA/TEOS) Pervaporation Membrane. The energy consumption includes the external heating and cooling required for the feed and permeate streams, as well as the electrical power associated with pumps for re-circulating feed and maintaining vacuum. The thermal energy requirement is significant (e.g., up to 2609 MJ/m³ of thermal energy) and is required to maintain the feed stream at 65 °C in recirculation mode. The electrical energy requirement is very small (<0.2 kWh/m³ of required at 65 °C feed temperature at steady state) with the vacuum pump contributing to the majority of the electrical energy. The energy required for the pervaporation process was also compared to other desalination processes such as Reverse Osmosis (RO), Multi-stage Flash (MSF), and Multiple Effect Distillation (MED). The electrical energy requirement for pervaporation is the lowest among these desalination technologies. However, the thermal energy needed for pervaporation is significant. Pervaporation may be attractive when the process is integrated with waste heat and heat recovery option and used in niche applications such as RO brine concentration or salt recovery.

Thin-layer chromatography combined with diode laser thermal vaporization inductively coupled plasma mass spectrometry for the determination of selenomethionine and selenocysteine in algae and yeast.

In this work we present a simple and cost-effective approach for the determination of selenium species in algae and yeast biomass, based on a combination of thin-layer chromatography (TLC) with diode laser thermal vaporization inductively coupled plasma mass spectrometry (DLTV ICP MS). Extraction of freeze-dried biomass was performed in 4M methanesulphonic acid and the selenium species were vaporized from cellulose TLC plates employing a continuous-wave infrared diode laser with power up to 4 W using a simple laboratory-built apparatus. Selenomethionine and selenocysteine were quantified with limits of detection 3 µg L-1 in a Se-enriched microalgae Chlorella vulgaris and yeast certified reference material SELM-1. Results delivered by TLC-DLTV ICP MS were consistent with those obtained by a routine coupling of high-performance liquid chromatography (HPLC) to ICP MS. In addition, the TLC approach is capable of analyzing extract containing even undiluted crude hydrolysates that could damage HPLC columns.

An integrated precipitation and ion-exchange chromatography process for antibody manufacturing: Process development strategy and continuous chromatography exploration.

In the past decades, research was carried out to find cost-efficient alternatives to Protein A chromatography as a capture step in monoclonal antibody (mAb) purification processes. In this work, polyethylene glycol (PEG) precipitation has shown promising results in the case of mAb yield and purity. Especially with respect to continuous processing, PEG precipitation has many advantages, like low cost of goods, simple setup, easy scalability, and the option to handle perfusion reactors. Nevertheless, replacing Protein A has the disadvantage of renouncing a platform unit operation as well. Furthermore, PEG precipitation is not capable of reducing high molecular weight impurities (HMW) like aggregates or DNA. To overcome these challenges, an integrated process strategy combining PEG precipitation with cation-exchange chromatography (CEX) for purification of a mAb is presented. This work discusses the process strategy as well as the associated fast, easy, and material-saving process development platform. These were implemented through the combination of high-throughput methods with empirical and mechanistic modeling. The strategy allows the development of a common batch process. Additionally, it is feasible to develop a continuous process. In the presented case study, a mAb provided from cell culture fluid (HCCF) was purified. The precipitation and resolubilization conditions as well as the chromatography method were optimized, and the mutual influence of all steps was investigated. A mAb yield of over 95.0% and a host cell protein (HCP) reduction of over 99.0% could be shown. At the same time, the aggregate level was reduced from 3.12% to 1.20% and the DNA level was reduced by five orders of magnitude. Furthermore, the mAb was concentrated three times to a final concentration of 11.9mg/mL.

A modified quick, easy, cheap, effective, rugged, and safe cleanup method followed by liquid chromatography-tandem mass spectrometry for the rapid analysis of perchlorate, bromate and hypophosphite in flour.

A selective, sensitive and useful method, based on modified QuEChERS cleanup combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the negative-ion electrospray ionization (ESI-) mode, was developed and validated for the simultaneous determination of three inorganic anions including perchlorate (ClO4-), bromate (BrO3-) and hypophosphite (H2PO2-) in flour. The extraction parameters and LC-MS/MS conditions were optimized by single-factor experiment and sorbent combination in modified QuEChERS clean-up was optimized through response surface analysis. Three target analytes were separated on a normal-phase Phenomenex Luna Silica (2) column (150mm×2.0mm, 5µm, 100Å) with the mobile phase of a mixture of 5mmol/L ammonium acetate water solution and acetonitrile, detected by MS/MS under multiple reaction monitoring and quantified by external standard method. The developed method was validated in terms of the sensitivity, linearity, accuracy and precision, and matrix effect. The method showed a good linearity (R2>0.999) for all analytes in their respective concentration ranges. The ILOQs and MLOQs for perchlorate, bromate and hypophosphite were 0.1, 0.5, 5.0µg/L and 2.0, 6.0, 60.0µg/L, respectively. The average recoveries of three target analytes from the negative samples spiked at three different concentrations were in a range from 84.6% to 104.9%. The intra-day precision (n=6) and inter-day precision (n=5) of the target analytes were in the ranges of 2.9%-6.9% and 6.4%-8.2%. The matrix effect of this method was observed between 0.83 and 1.17 and was acceptable. The validated method was successfully applied to determine the concentrations of these inorganic anions in flour. Results found that perchlorate and hypophosphite were detected in 33 out of 50 and 7 out of 50 flour samples.

Highly Specific and Wide Range NO2 Sensor with Color Readout.

We present a simple and inexpensive method to implement a Griess-Saltzman-type reaction that combines the advantages of the liquid phase method (high specificity and fast response time) with the benefits of a solid implementation (easy to handle). We demonstrate that the measurements can be carried out using conventional RGB sensors; circumventing all the limitations around the measurement of the samples with spectrometers. We also present a method to optimize the measurement protocol and target a specific range of NO2 concentrations. We demonstrate that it is possible to measure the concentration of NO2 from 50 ppb to 300 ppm with high specificity and without modifying the Griess-Saltzman reagent.

Facile and green fabrication of cation exchange membrane adsorber with unprecedented adsorption capacity for protein purification.

Fabricating membrane adsorbers with high adsorption capacity and appreciable throughput for the separation and purification of protein products is challenging in biomedical and pharmaceutical industries. Herein, we report the synthesis of a novel membrane adsorber by functionalizing a nylon microfiltration membrane with alginate dialdehyde (ADA) followed by sulphonic addition, without any solvent usage, and its successful application in the purification of lysozyme. Taking advantage of abundant dual cation exchange (CEX) groups on sulphonic-ADA (S-ADA) ligands, this novel S-ADA-nylon membrane adsorber showed an unprecedented static binding capicity of 286mg/mL for lysozyme adsorption. Meanwhile, the prepared membrane adsorber could be easily regenerated (complete protein elution) under mild conditions and be reused at least for five times. Featured with a unique selectivity, the S-ADA-nylon membrane also captured lysozyme from chicken egg white solution with a high purity (100%) and a high recovery of 98%. The purified lysozyme showed similar specific activity as commercial product. The present work provides a facile, green and low-cost approach for the preparation of high-performance membrane adsorbers, which has a great potential in protein production.

Comparison of micellar extraction combined with ionic liquid based vortex-assisted liquid-liquid microextraction and modified quick, easy, cheap, effective, rugged, and safe method for the determination of difenoconazole in cowpea.

Two simple sample pretreatment for the determination of difenoconazole in cowpea was developed including micellar extraction combined with ionic liquid based vortex-assisted liquid-liquid microextraction (ME-IL-VALLME) prior to high performance liquid chromatography (HPLC), and modified quick, easy, cheap, effective, rugged, and safe method (QuEChERS) coupled with HPLC-MS/MS. In ME-IL-VALLME method, the target analyte was extracted by surfactant Tween 20 micellar solution, then the supernatant was diluted with 3mL water to decrease the solubility of micellar solution. Subsequently, the vortex-assisted liquid-liquid microextraction (VALLME) procedure was performed in the diluted extraction solution by using the ionic liquid of 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIM]PF6) as the extraction solvent and Tween 20 as an emulsifier to enhance the dispersion of the water-immiscible ionic liquid into the aqueous phase. Parameters that affect the extraction have been investigated in both methods Under the optimum conditions, the limits of quantitation were 0.10 and 0.05mgkg-1, respectively. And good linearity was achieved with the correlation coefficient higher than 0.9941. The relative recoveries ranged from 78.6 to 94.8% and 92.0 to 118.0% with the relative standard deviations (RSD) of 7.9-9.6% and 1.2-3.2%, respectively. Both methods were quick, simple and inexpensive. However, the ME-IL-VALLME method provides higher enrichment factor compared with conventional QuEChERS method. The ME-IL-VALLME method has a strong potential for the determination of difenoconazole in complex vegetable matrices with HPLC.

Current status of bioanalytical outsourcing: a pharma perspective.

Eric Woolf, Ajai Chaudhary and Sheila Breidinger from Merck, spoke to Sankeetha Nadarajah (Managing Commissioning Editor, Bioanalysis) about outsourcing strategy implementation.

Outsourcing bioanalytical services at Janssen Research and Development: the sequel anno 2017.

The strategy of outsourcing bioanalytical services at Janssen has been evolving over the last years and an update will be given on the recent changes in our processes. In 2016, all internal GLP-related activities were phased out and this decision lead to the re-orientation of the in-house bioanalytical activities. As a consequence, in-depth experience with the validated bioanalytical assays for new drug candidates is currently gained together with the external partner, since development and validation of the assay and execution of GLP preclinical studies are now transferred to the CRO. The evolution to externalize more bioanalytical support has created opportunities to build even stronger partnerships with the CROs and to refocus internal resources. Case studies are presented illustrating challenges encountered during method development and validation at preferred partners when limited internal experience is obtained or with introduction of new technology.