Ten Basic Rules of Antibody Validation

The quality of research antibodies is an issue for decades. Although several papers have been published to improve the situation, their impact seems to be limited. This publication makes the effort to simplify the description of validation criteria in a way that the occasional antibody user is able to assess the validation level of an immunochemical reagent. A simple, 1-page checklist is supplied for the practical application of these criteria.

Chitosan Membrane Embedded With ZnO/CuO Nanocomposites for the Photodegradation of Fast Green Dye Under Artificial and Solar Irradiation

Fast Green (FCF) dye is commonly used in both cytology and histology applications. Previous studies have found that it can cause mutagenic and tumorigenic effects in experimental human and animal populations. It can also be a source of skin, eye, respiratory, and digestive irritation. The purpose of this study was to examine the use of thin film membranes to degrade FCF. A thin film membrane of chitosan (CS) was fabricated and subsequently filled with zinc oxide nanoparticles (ZnO) or ZnO/CuO-heterostructured nanocomposites. The CS membrane was used as a matrix, and the nanomaterials were used as photocatalysts. The prepared membranes were characterised by four analytical techniques: atomic force microscopy, scanning electron microscopy, X-ray diffraction, and energy-dispersive X-ray analyses. The photocatalytic activity of the fabricated membranes was evaluated by performing experiments in which aqueous solutions of FCF dye that contained the fabricated membrane were irradiated with solar light or UV light. The photodegradation percentage was spectrophotometrically determined by monitoring the maximum wavelengths (λmax) of FCF at 623 nm for different irradiation times. The decolourisation percentages of the dye under solar light were 57.90% and 60.23% using the CS-ZnO and CS-ZnO/CuO membranes, respectively. When UV light irradiation was employed as the source of irradiation, the photodegradation percentages of FCF were 71.45% and 91.21% using the CS-ZnO and CS-ZnO/CuO membranes, respectively. These results indicated that the best photocatalytic system for the degradation of FCF dye was CS-ZnO/CuO membrane in combination with UV light irradiation. The study also found that it was easy to separate the prepared membranes after the reaction without the need for a centrifuge or magnet. The results demonstrate the potential for CS-ZnO and CS-ZnO/CuO membranes for use as effective sorbents during the process of photodegradation of harmful dyes within waste water recycling practices.

In Vitro Studies of the Antimicrobial and Free-Radical Scavenging Potentials of Silver Nanoparticles Biosynthesized From the Extract of Desmostachya bipinnata

The aim of this study was to perform green synthesis of silver nanoparticles (AgNPs) from the leaf extract of Desmostachya bipinnata (Dharba), a medicinally important herb which is widely used across India. Synthesized AgNPs were analyzed by UV-Visible spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDAX). The results have confirmed that green synthesis of AgNPs leads to the fabrication of sphere-shaped particles with a diameter of 53 nm. Furthermore, these AgNPs were subjected to antioxidant and antimicrobial studies against gram-negative and gram-positive bacteria, where AgNPs at a concentration of 20 mg/mL showed highest zone of inhibition. Synthesized AgNPs were evaluated for their antioxidant activity by 1, 1-diphenyl-2-picryl hydrazyl radical (DPPH), H2O2, and superoxide inhibiting assays; increasing concentration has showed increase in scavenging ability. Cell toxicity was assessed on HepG2 cell lines, and synthesized nanoparticles at a concentration of 128 μg/mL produced significant reduction in viability of Hep cells ( P < .05). The availability of Dharba throughout the year and the eco-friendly approach in the synthesis of AgNPs coupled with bioactivity has demonstrated its potential as a novel biomaterial which can be used for various biomedical applications.

Analytical Methods for the Determination of Mineral Oil Saturated Hydrocarbons (MOSH) and Mineral Oil Aromatic Hydrocarbons (MOAH)—A Short Review

Mineral oils (such as paraffinum liquidum or white oil), which consist of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH), are widely applied in various consumer products such as medicines and cosmetics. Contamination of food with mineral oil may occur by migration of mineral oil containing products from packaging materials, or during the food production process, as well as by environmental contamination during agricultural production. Considerable analytical interest was initiated by the potential adverse health effects, especially carcinogenic effects of some aromatic hydrocarbons. This article reviews the history of mineral oil analysis, starting with gravimetric and photometric methods, followed by on-line-coupled liquid chromatography with gas chromatography and flame ionization detection (LC-GC-FID), which still is considered as gold standard for MOSH-MOAH analysis. Comprehensive tables of applications in the fields of cosmetics, foods, food contact materials, and living organisms are provided. Further methods including GCxGC-MS methods are reviewed, which may be suitable for confirmation of LC-GC-FID results and identification of compound classes. As alternative to chromatography, nuclear magnetic resonance (NMR) spectroscopy has recently been suggested for MOSH-MOAH analysis, especially with the possibility of detecting only the toxicologically relevant aromatic rings. Furthermore, NMR may offer potential as rapid screening especially with low-field instruments usable for raw material control.

An Accurate Assessment of Docosahexaenoic Acid in Laying Hen Serum for Regulatory Studies

Diets rich in omega-3 fatty acids (n-3 FA) have been associated with several health benefits. With the increased interest in n-3 FA both scientifically and societally, the accurate detection of such analytes has become increasingly important. Recently, tandem mass spectrometry (MS/MS) with electrospray ionization interface (ESI), hyphenated to both gas chromatography (GC) and liquid chromatography (LC), has become a valuable tool in the detection of docosahexaenoic acid (DHA). Liquid chromatography-electrospray ionization interface-tandem mass spectrometry methods have been developed for the determination of DHA in canine and poultry species. The objective of this article is to investigate whether LC-ESI-MS/MS is fit for purpose for the determination of DHA in laying hen serum. The disclosure of this work will be beneficial for researchers investigating poultry enrichment for regulatory and toxicological studies. The method was found to be linear over the range. Precision and accuracy results met acceptance criteria and the Limit of Quantitation (LOQ) was established as 1 µg/mL. Recoveries of DHA were obtained for quality control samples and stability studies were performed. The results of the verification study complimented those of the validation study. In summation, the method was established as fit for purpose for measuring total DHA in laying hen serum.

Quality by Design–Applied Liquid Chromatography-Tandem Mass Spectrometry Determination of Enzalutamide Anti-Prostate Cancer Therapy Drug in Spiked Plasma Samples

This research article presents the Quality by Design (QbD)–finalised conditions for a method that uses liquid chromatography-tandem mass spectrometry for the determination of concentration of enzalutamide (ENZ), an atypical anticancer drug, in a drug formulation and in spiked plasma samples. Critical process attributes (CPA) considered to be the influential parameters in separation, identification, and quantification processes by ultrahigh-performance liquid chromatography-electrospray ionisation-tandem mass spectrometry (UHPLC-ESI-MS/MS) were organic content, buffer strength, pH modifier, flow rate, spray voltage, sheath gas, and auxiliary gas that alter critical analytical attributes, such as retention time (R1) and area (R2). These factors were evaluated first in a factorial design (Taguchi orthogonal array design) and then extensively in a central composite design (CCD) to zero-in on the mobile phase for the quantification of ENZ standard drug and along with its internal standard (ENZIS) in spiked plasma samples and in formulation. Pareto chart from initial factorial design (Taguchi orthogonal array design) model suggested which of the CPA factors should be given the weightage, that is, to be exhaustively analysed in the CCD and response surface analysis. The elaborated parameters proposed by World Health Organization were studied by method validation, ie, selectivity, linearity, accuracy, precision repeatability system-suitability tests, method robustness/ruggedness, sensitivity, and stability. The strategy followed gives an insight on the development of a robust QbD-compliant quantitative UHPLC-ESI-MS/MS method for ENZ drug containing plasma samples (spiked).

Synthesis and Optical Properties of Pentamethine Cyanine Dyes With Carboxylic Acid Moieties

Cyanine dyes possessing carboxylic acid groups have been used in many different fields of study. The acid groups can act as handles for bioconjugation or as metal chelators. Several pentamethine cyanine dyes with propionic acid handles were synthesized and their optical properties were studied to determine their usefulness as fluorescent probes. The optical properties studies performed include the absorbance and emission maxima values as well as the calculation of quantum yield and molecular brightness levels. Molecular models were also calculated to help analyze the dyes’ behavior and were compared with similar dyes with varying alkyl chain lengths replacing the acid moieties.

Comparison of 3 Derivatization Methods for the Analysis of Amphetamine-Related Drugs in Oral Fluid by Gas Chromatography-Mass Spectrometry

The heptafluorobutyric anhydride (HFBA), pentafluoropropionic anhydride (PFPA), and trifluoroacetic anhydride (TFAA) are compared as derivatizing reagents to use as the optimal method for the analysis of 10 amphetamines and cathinones in oral fluid. The target compounds were amphetamine (AMP), methamphetamine (MA), 4-methylamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxy- N-ethylamphetamine (MDEA), cathinone (CAT), methcathinone, mephedrone, and ephedrine. Amphetamine-D5, MA-D5, MDA-D5, MDMA-D5, and MDEA-D5 use as internal standards (IS). The analytes and IS were extracted from 0.5 mL of oral fluid by ethyl acetate in the presence of NaOH (0.1 N) as the base and then the dried extracts were derivatized with HFBA, PFPA, or TFAA at 70°C for 30 minutes. The limits of quantification based on signal-to-noise ratios ≥10 were ranged between 2.5 and 10 ng/mL. The calibration graphs were linear in the range of 5 or 10 to 1000 ng/mL for all analytes. Based on sensitivity, the PFPA is proved to be the best for derivatization of the target compounds prior to gas chromatography-mass spectrometry analysis.

Colorimetric Detection Based on Localised Surface Plasmon Resonance Optical Characteristics for the Detection of Hydrogen Peroxide Using Acacia Gum–Stabilised Silver Nanoparticles

The use of nanoparticles in sensing is attracting the interest of many researchers. The aim of this work was to fabricate Acacia gum–stabilised silver nanoparticles (SNPs) using green chemistry to use them as a highly sensitive and cost-effective localised surface plasmon resonance (LSPR) colorimeter sensor for the determination of reactive oxygen species, such as hydrogen peroxide (H2O2). Silver nanoparticles were fabricated by the reduction of an inorganic precursor silver nitrate solution (AgNO3) using white sugar as the reducing reagent and Acacia gum as the stabilising reagent and a sonication bath to form uniform silver nanoparticles. The fabricated nanoparticles were characterised by visual observation, ultraviolet-visible (UV-Vis) spectrophotometry, transmission electron microscopy (TEM) analysis, energy-dispersive X-ray spectroscopy (EDAX), thermogravimetric analysis (TGA), and Fourier transform infrared spectroscopy (FT-IR). The TEM micrographs of the synthesised nanoparticles showed the presence of spherical nanoparticles with sizes of approximately 10 nm. The EDAX spectrum result confirmed the presence of silver (58%), carbon (30%), and oxygen (12%). Plasmon colorimetric sensing of H2O2 solution was investigated by introducing H2O2 solution into Acacia gum–capped SNP dispersion, and the change in the LSPR band in the UV-Vis region of spectra was monitored. In this study, it was found that the yellow colour of Acacia gum–stabilised SNPs gradually changed to transparent, and moreover, a remarkable change in the LSPR absorbance strength was observed. The calibration curve was linear over 0.1–0.00001 M H2O2, with a correlation estimation ( R2) of .953. This was due to the aggregation of SNPs following introduction of the H2O2 solution. Furthermore, the fabricated SNPs were successfully used to detect H2O2 solution in a liquid milk sample, thereby demonstrating the ability of the fabricated SNPs to detect H2O2 solution in liquid milk samples. This work showed that Acacia gum–stabilised SNPs may have the potential as a colour indicator in medical and environmental applications.

Review of Current Applications of Immunohistochemistry in Pediatric Nonneoplastic Gastrointestinal, Hepatobiliary, and Pancreatic Lesions

Immunohistochemical (IHC) stains are widely used by pathologists for a variety of considerations in the diagnostic workup of pediatric nonneoplastic lesions in gastrointestinal (GI), hepatic, biliary, and pancreatic lesions. The pathologic changes cover a wide range and types of presentations, including inflammatory (bacterial and viral), metaplastic, posttransplant lymphoproliferative, autoimmune, metabolic, degenerative, developmental, and genetic conditions, among others. The everyday practical value of IHC stains covers primary identification, confirmation, differential, and/or exclusionary roles in the hands and eyes and minds of the practitioners. This article is intended to review and discuss the currently available IHC stains for a variety of pediatric GI, hepatobiliary, and pancreatic lesions as encountered in the day-to-day practice of pathologists and clinicians. It reflects the most recent methods and types of IHC stains with the stated aim of helping to provide a quick reference for diagnostic considerations and thereby facilitate the workup of a broad range of GI and related conditions in a pediatric population. The tables provide a handy reference on a wide range of IHC stains for commonly encountered lesions covering a variety of pediatric GI, hepatobiliary, and pancreatic conditions that are amenable to light microscopic diagnostic interpretation.