Standard Addition Approach Research Articles

Smart mathematically filtered UV spectroscopic methods for quality assurance of rosuvastatin and valsartan from formulation

Abstract Valsartan and rosuvastatin together in a binary form have been utilized to reduce hypertension and hyperlipidemia to control cardiovascular complications. This study depicts the simple three mathematically manipulated UV spectroscopic techniques for the estimation of rosuvastatin and valsartan in the formulation. The first method is simple UV absorption at 310 nm by RST and the first derivatization method for VTN. Determining the magnitude difference of a ratio spectrum at two identified wavelengths is the second approach, and determination of the magnitude of the first derivatives of the ratio spectra of RST and VTN constitute the third technique. The selection of wavelengths, divisor concentrations, and peak amplitudes were optimized and validated. The straight line was constructed in the range of 1–30 and 2–25 µg/ml for RST and VST by the normal and first derivatization method. By using the magnitude difference and magnitude of first derivative ratio spectra approaches, the concentrations of 1–12 and 2–25 µg/ml for RST and VTN, respectively, displayed a straight line. The limit of quantification was less than 1 µg/ml for RST and less than 2 µg/ml for VTN. It was eventually found that the accuracy, expressed as a percentage recovery, ranged between 98.94 and 99.55% for RST and 100.36 and 101.08% for VTN. The % RSD did not exceed 1.82 and 1.91 for RST and VTN, respectively. The three techniques were used to accurately measure RST and VTN in their binary formulations and physically mixed solutions, and the results were statistically compared to the previously published HPLC technique. The outstanding recovery achieved by using the authentic standard addition approach validated the methods’ supplemental accurateness. The Analytical Greenness and Red Green Blue procedures verified the eco-friendliness of the suggested UV spectroscopic approaches, which were also found to be superior to the documented HPLC methods.

Fabrication and characterization of model wireless biosensor-based electrochemical impedance spectroscopy (EIS) for detecting HER2 in plasma as therapeutics

PurposeEarly prediction of any type of cancer is important for the treatment of this type of disease, therefore, our target to evaluate whether monitoring early changes in plasma human epidermal growth factor receptor 2 (HER2) levels (using EIS), could help in the treatment of breast cancer or not? Human epidermal growth factor receptor 2 (HER2) overexpression is an important biomarker for treatment selection in earlier stages of cancers. The combined detection of the HER2 gene in plasma for blood cancer provides an important reference index for the prognosis of metastasis to other tissues. For this purpose, the authors fabricated and characterized a model wireless biosensor-based electrochemical impedance spectroscopy (EIS) for detecting HER2 plasma as therapeutics.Design/methodology/approachMost sensors generally are fabricated based on a connection between component of the sensors and the external circuits through wires. Although these types of sensors provide suitable sensitivities and also quick responses, the connection wires can be limited to the sensing ability in various devices approximately. Therefore, the authors designed a wireless sensor, which can provide the advantages of in vivo sensing and also long-distance sensing, quickly.FindingsThe biosensor structure was designed for detection of HER2, HER3 and HER-4 from lab-on-chip approach with six units of screen-printed electrode (SPE), which is built of an electrochemical device of gold/silver, silver/silver or carbon electrodes. The results exhibited that the biosensor is completely selective at low concentrations of the plasma and HER2 detection via the standard addition approach has a linearity plot, therefore, by using this type of biosensors HER2 in plasma can be detected.Originality/valueThis is then followed by detecting HER2 in real plasma using standard way which proved to have great linearity (R2 = 0.991) proving that this technique can be used to detect HER2 solution in real patients.

Indicator displacement-based colorimetric assay for dibutyl phthalate in pharmaceutical products with titanium(IV)-pyridylazo resorcinol (PAR)

Taking advantage of the competitive binding affinity towards Ti(IV) between 4-(2-pyridylazo) resorcinol (PAR) and phthalate, a simple indicator displacement (ID)–based colorimetric assay was designed for indirect determination of a well-known phthalic acid ester, dibutyl phthalate (DBP). The indicator PAR and Ti(IV) formed a purplish-red-colored Ti(IV)-PAR complex (λmax = 540 nm) at a 1:1 ratio. In the presence of pre-hydrolyzed DBP, colorless complex formation of phthalate ion (emerging from alkaline hydrolysis of DBP) with Ti(IV) resulted in a hypsochromic shift in absorbance maximum, accompanying a color change from purplish-red to yellowish-orange (λmax = 390 nm) by the release of PAR from Ti(IV)-PAR system. Based on this mechanism, the linear response range of the system for DBP was found to lie between 0.16 and 0.37 mmol L–1 with an experimental detection limit of 11.6 µmol L–1. The recommended Ti(IV)-PAR system was successfully applied to DBP-containing pharmaceutical products (as real sample) after a simple clean-up process for removing possible water-soluble interferents. The analytical results obtained from the recommended method (by applying the standard addition approach) and the reference liquid chromatography-tandem mass spectrometric (LC–MS/MS) method were statistically compared using DBP-extract of the drug samples. Consequently, a simple and selective colorimetric ID strategy was proposed for the analysis of DBP in pharmaceuticals for the first time.

Quantitative analysis of residual butylated hydroxytoluene and butylated hydroxyanisole in Salmo salar, milk, and butter by liquid chromatography–tandem mass spectrometry

Butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) are two commonly used antioxidants with potential health risks associated with excessive intake from multiple sources. Several countries have implemented strict regulations to curb these risks. This study presents a simple LC–MS/MS method for estimating BHT and BHA levels in Salmo salar, butter, and milk. To mitigate any potential interference from the three complex matrices with the ionisation of the target analytes, the method utilised the standard addition approach. The mobile phase used to elute the analytes consisted of 0.1 % formic acid in a mixture of water and acetonitrile (25:75 v/v). Both antioxidants were detected in negative ionisation mode. BHT was identified through single-ion monitoring at a mass-to-charge ratio (m/z) of 219.4, while BHA was detected using multiple-reaction monitoring, with a transition from m/z 164.0 to 149.0. The environmental assessment of the applied procedures verified that the approach is eco-friendly.

Determination of Phosphorus Content in Cola Drinks by 31P NMR Spectroscopy Using External Calibration and Standard Addition Approaches

Determination of Phosphorus Content in Cola Drinks by ³¹P NMR Spectroscopy Using External Calibration and Standard Addition Approaches

Investigation of use of hydrophilic/hydrophobic NADESs for selective extraction of As(III) and Sb(III) ions in vegetable samples: Air assisted liquid phase microextraction and chemometric optimization

In this paper, a green, cost-effective sample preparation method based on air assisted liquid phase microextraction (AA-LPME) was developed for the simultaneous extraction of As(III) and Sb(III) ions from vegetable samples using hydrophilic/hydrophobic natural deep eutectic solvents (NADESs). Central composite design was used for the optimization of extraction factors including NADES volume, extraction cycle, pH, and curcumin concentration. Limits of detection for As(III) and Sb(III) were 1.5 ng L−1 and 0.06 ng L−1, respectively. Working ranges for As(III) and Sb(III) were 0.2–300 ng L−1 (coefficient of determination (R2 = 0.9978) and 5–400 ng L−1 (R2 = 0.9996), respectively. Relative standard deviations for As(III) and Sb(III) were 2.2–2.8% and 2.9–3.2%, respectively. Enrichment factor of the method was 184 for As(III) and 172 for Sb(III). The accuracy and precision of the AA-NADES-LPME method were investigated by intraday/interday studies and standard reference material analysis, respectively. Finally, the AA-NADES-LPME method was successfully applied to microwave digested vegetable samples using the standard addition approach and acceptable recoveries were achieved.

Optimization of vortex-assisted magnetic nanofluid based liquid phase microextraction for fast, green and reliable extraction of ibuprofen from real samples: Multivariate approach

In this research, the vortex-assisted magnetic nanofluid based liquid phase microextraction (VA-MNF-LPME) procedure was optimized using Box–Behnken Design (BBD) for the selective and reliable extraction of ibuprofen from aqueous solutions. Three MNFs were prepared and tested for the selective extraction of ibuprofen. Ultraviolet–visible spectrophotometry (UV–Vis) was used to determine the amount of ibuprofen and measurements were made at a wavelength of 238 nm. The effects of pH, MNF volume, vortex time, volume of back extraction solvent were optimized using BBD. Separation of the MNF phase containing the ibuprofen in aqueous solution was achieved using a neodymium magnet without the need for centrifugation step. Using optimized parameters (pH = 3.4, 690 μL of MNF, 100 s vortex and 230 μL of ethanol), the detection limit and working range of the V-MNF-LPME procedure were 0.6 ng mL−1 and 2–180 ng mL−1, respectively. Relative standard deviations and recovery (for 25 ng mL−1 of ibuprofen solution, N = 5) were 2.8% and 94.1%. Key validation parameters of the method were studied in detail and calculated under optimized conditions. The VA-MNF-LPME procedure was successfully applied to some drugs, baby lettuce and grape leaf samples in ten stages with a standard-addition approach, and quantitative recoveries were obtained (91.4–98.7%).

Novel method for chlorine determination in biofuel lipid feedstock by coupling a total sample introduction system (hTISIS) to an inductively coupled plasma tandem mass spectrometry (ICP-MS/MS)

BackgroundChlorine concentrations above 1 mg kg−1 in lipid feedstocks for biofuel production can generate corrosion issues in the different refining units as well as catalyst deactivation by clogging or fouling. To reach accurate analyses by inductively coupled plasma (ICP) techniques at low concentration levels, dilution in organic solvents rises as a simpler and more straightforward sample preparation methodology than conventional sample decomposition procedures (e.g., microwave-assisted acid digestion). However, matrix effects and the impact of the Cl chemical form on the signal must be overcome to obtain accurate results. ResultsIn this work, the high temperature torch integrated sample introduction system (hTISIS) operated at 350 °C is coupled to an inductively coupled plasma tandem mass spectrometry (ICP-MS/MS) for the determination of Cl in lipid biofuel feedstock samples diluted in xylene and these results are compared with those reported by a conventional sample introduction system. Under optimal conditions of the hTISIS-ICP-MS/MS configuration, matrix effects are efficiently overcome (recovery values ranged from 101 to 104%) as well as the effect of the Cl chemical form on the signal for 6 organochloride compounds. Thus, an external calibration approach can be set to carry out the quantification of this element in real samples. The method is successfully validated, obtaining a good agreement in the Cl concentration reported in a standard reference material (SRM NIST 1634c) and also by comparing the concentration results obtained by external calibration and standard addition approaches in two biofuel feedstock samples. Significance and noveltyThe hTISIS coupled to an ICP-MS/MS system is used for the first time to overcome not only matrix effects but also the impact of the Cl chemical form in biofuel feedstock samples. This novel method, with a limit of quantification (LOQ) of 7.1 μg kg−1, give access to an accurate Cl determination in all kind of lipid feedstocks for clean fuel production.

A New Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry Method to Monitor 1-Amino-4-Methyl-Piperazine and 1-Methyl-4-Nitroso-Piperazine in Rifampicin Hydrolysis: A Standard Addition Approach to Quantify Nitrosamine in Rifampicin

1-amino-4-methyl-piperazine (AMP) is both a starting material for the synthesis of rifampicin (RIF) and a degradation product of RIF hydrolysis. 1-methyl-4-nitroso-piperazine (MNP) is an oxidation product of AMP as well as a potentially genotoxic N-nitrosamine. The EMA and FDA have approved an ad interim limit of 5 ppm for MNP in RIF drug products. As in-house methods for the analysis of MNP in RIF use a wide range of conditions for mobile phases and sample diluents, we decided to investigate whether these conditions affect the formation of MNP and AMP. A UHPLC-MS/MS method was developed to simultaneously quantify AMP and MNP during RIF hydrolysis in buffered aqueous solutions at different pH levels. Analyses were performed in MRM mode; separations were carried out on an InfinityLab Poroshell HPH-C18 (100 mm, 2.1 mm i.d., particle size 1.9 μm). In aqueous RIF solutions, the content of AMP and MNP increases with time; at different pHs, the concentration of AMP increases much faster in acidic than in basic solutions; and the increase in MNP can be reduced by the addition of ascorbic acid. To avoid an overestimation of MNP, water should not be used as a diluent in RIF sample preparations. Methanol is a more suitable diluent than water. A standard addition method has been validated for the quantification of MNP in RIF drug substances.

Comparing Sensor Fusion and Multimodal Chemometric Models for Monitoring U(VI) in Complex Environments Representative of Irradiated Nuclear Fuel.

Optical sensors and chemometric models were leveraged for the quantification of uranium(VI) (0-100 μg mL-1), europium (0-150 μg mL-1), samarium (0-250 μg mL-1), praseodymium (0-350 μg mL-1), neodymium (0-1000 μg mL-1), and HNO3 (2-4 M) with varying corrosion product (iron, nickel, and chromium) levels using laser fluorescence, Raman scattering, and ultraviolet-visible-near-infrared absorption spectra. In this paper, an efficient approach to developing and evaluating tens of thousands of partial least-squares regression (PLSR) models, built from fused optical spectra or multimodal acquisitions, is discussed. Each PLSR model was optimized with unique preprocessing combinations, and features were selected using genetic algorithm filters. The 7-factor D-optimal design training set contained just 55 samples to minimize the number of samples. The performance of PLSR models was evaluated by using an automated latent variable selection script. PLS1 regression models tailored to each species outperformed a global PLS2 model. PLS1 models built using fused spectra data and a multimodal (i.e., analyzed separately) approach yielded similar information, resulting in percent root-mean-square error of prediction values of 0.9-5.7% for the seven factors. The optical techniques and data processing strategies established in this study allow for the direct analysis of numerous species without measuring luminescence lifetimes or relying on a standard addition approach, making it optimal for near-real-time, in situ measurements. Nuclear reactor modeling helped bound training set conditions and identified elemental ratios of lanthanide fission products to characterize the burnup of irradiated nuclear fuel. Leveraging fluorescence, spectrophotometry, experimental design, and chemometrics can enable the remote quantification and characterization of complex systems with numerous species, monitor system performance, help identify the source of materials, and enable rapid high-throughput experiments in a variety of industrial processes and fundamental studies.

Spectrofluorimetric determination of vitamin D in the serum of autistic and healthy children using functionalized graphene quantum dots

Vitamin D is one of the most essential nutrients for brain development, and deficiencies during pregnancy and early childhood development might be associated with autism. Regular monitoring of serum 25-hydroxyvitamin D3 level could help in early diagnosis and therapy. Analytical measurement of serum 25-hydroxyvitamin D3 level using the traditional matrix-matched calibration technique yields inaccurate results due to absence of serum matrix free from 25-hydroxyvitamin D3. The aim of this work was to develop a validated spectrofluorimetric methodology based on the standard addition approach for quantifying 25-hydroxyvitamin D3 levels in real serum samples of autistic children. The spectrofluorimetric methodology utilizes functionalized graphene quantum dots as a fluorescent probe for selective quantification of 25-hydroxyvitamin D3 level, which is based on measuring the quenching properties of 25-hydroxyvitamin D3 on a fluorescent probe. The standard addition approach exhibits a minimal matrix interference since it identically utilizes the same matrix of each study sample for creating its own calibration curve. The method was validated using the guidelines outlined in ICH M10 draft for endogenous compounds quantification. The method was successfully applied for quantifying the serum 25-hydroxyvitamin D3 levels in autistic and healthy children, and autistic children had significantly lower serum 25-hydroxyvitamin D3 levels (with a mean ± SD of 23.80 ± 17.19) when compared to healthy children (with a mean ± SD of 50.13 ± 18.74, P < 0.001). These results suggested an association between vitamin D deficiency and autism.

Standard addition method for rapid, cultivation-independent quantification of Legionella pneumophila cells by qPCR in biotrickling filters.

Cultivation-independent molecular biological methods are essential to rapidly quantify pathogens like Legionella pneumophila (L. pneumophila) which is important to control aerosol-generating engineered water systems. A standard addition method was established to quantify L. pneumophila in the very complex matrix of process water and air of exhaust air purification systems in animal husbandry. Therefore, cryopreserved standards of viable L. pneumophila were spiked in air and water samples to calibrate the total bioanalytical process which includes cell lysis, DNA extraction, and qPCR. A standard addition algorithm was employed for qPCR to determine the initial concentration of L. pneumophila. In mineral water, the recovery rate of this approach (73%-134% within the concentration range of 100-5000 Legionella per mL) was in good agreement with numbers obtained from conventional genomic unit (GU) calibration with DNA standards. In air samples of biotrickling filters, in contrast, the conventional DNA standard approach resulted in a significant overestimation of up to 729%, whereas our standard addition gave a more realistic recovery of 131%. With this proof-of-principle study, we were able to show that the molecular biology-based standard addition approach is a suitable method to determine realistic concentrations of L. pneumophila in air and process water samples of biotrickling filter systems. Moreover, this quantification strategy is generally a promising method to quantify pathogens in challenging samples containing a complex microbiota and the classical GU approach used for qPCR leads to unreliable results.

A Microfluidic Paper-based Analytical Device for Simultaneous Measurement of Albumin, Creatinine and Uric Acid in Urine based on Standard Addition Method

This work describes a simple method for the simultaneous measurement of urinary albumin, creatinine and uric acid using a standard addition approach on a microfluidic paper-based analytical device (µPAD). Hydrophobic barrier of the µPAD was created by stamping indelible ink onto a filter paper. The µPAD was designed with a flower-liked configuration. After aliquoting urine to a central sample zone, the sample flowed to ten surrounding channels, namely “the inner channels”, where the blank (water) or standard solutions had been added. The inner channels were connected to circular reagent zones where reagents had been immobilized. Tetrabromophenolphthalein ethyl ester, alkaline picrate, and a mixed solution of ferric chloride and ferric cyanide were used as the chromogenic reagents for the colorimetric detections of albumin, creatinine, and uric acid, respectively. Each reagent zone was linked with a circular detection zone through a second channel, namely “the outer channels”. The blue-, orange- and greenish-blue colored products were observed in the detection zones for the measurement of albumin, creatinine, and uric acid, correspondingly. A digital image of the µPAD was captured with a mobile phone. The color intensities were evaluated by ImageJTM and were employed for the quantitative analyses. The standard addition calibrations were found to be linear (r2 &gt; 0.99) for the spiked analyte concentrations up to 100 mg L-1 albumin, 1000 mg L-1 creatinine, and 50 mg dL-1 uric acid. The paper platform provided high precision (RSD &lt; 5 %) and good analytical recovery (91.8-109.7 %). Under paired t-test, the results obtained by the developed µPAD and the validating methods were not significantly different at 95 % confidence (albumin: tstat = -0.130, tcri = 3.182, creatinine: tstat = -0.133, tcri = 3.182, and uric acid: tstat = 1.119, tcri = 3.182).

Formation of plasmonic silver nanoparticles by glucosamine reduction: Application to a colorimetric sensor for determination of glucosamine in its pharmaceutical preparations

The purpose of this study is to develop a novel method for synthesizing silver nanoparticles using glucosamine as reducing agent and to utilize the developed method for colorimetric detection and quantitative determination of the non-chromophoric drug, glucosamine. Silver nanoparticles are prepared by reducing 0.02 mol/L silver nitrate by glucosamine in 0.075 mol/L ammonia and stabilizing the nanoparticles with 0.1% polyvinylpyrrolidone and the mixture is heated at 90 °C for 5 min. The prepared silver nanoparticles dispersed in water exhibit a bright yellow color due to a localized surface plasmon resonance band at 412 nm. The principle of glucosamine sensing is based on measuring the intensity of the surface plasmon resonance band at 412 nm which is directly proportional to the concentration of glucosamine with a linearity range (1 – 9 μg/mL), limit of detection 0.33 μg/mL and limit of quantitation 1.0 μg/mL. The proposed method was validated according to the ICH guidelines, and it was found to be accurate, precise, selective, and robust. The method was applied for determination of glucosamine in Joflex® capsules using the standard addition approach with mean % recovery ± standard deviation of 100.077 ± 1.786. The method is simple, rapid, and cost-effective and can be used for determination of glucosamine in bulk and in its pharmaceutical preparations.

Fluorescence-based absolute quantification of near-infrared probes in tissue extracts for biodistribution analyses

In recent years, significant progress has been made in the development of fluorescent contrast agents for clinical applications. For the development of a fluorescent probe, it is crucial to evaluate its safety profile, including biodistribution. Specific methods need to be developed for the absolute quantification of fluorescent probes in tissue specimens from animals administered with test compounds in the framework of biodistribution/efficacy/toxicity studies. Here, we describe a new method for the absolute quantification of fluorescent probes in tissue specimens from animals administered with compounds that have absorption and emission wavelength in the Near-Infrared region (600–800 nm). The protocol is based on the standard addition approach in order to minimize the interference of the matrix on the analyte signal causing inaccuracy in the absolute determination of the concentration. The measurement of the fluorescence intensity is done via a microplate reader. The method has been fully validated and applied for the quantification of a fluorescence-guided surgery targeted contrast agent in a Good Laboratory Practice (GLP) biodistribution study. Results clearly demonstrate that this procedure is fully applicable in a preclinical setting and that it overcomes common issues associated with fluorescence signal quantification in tissue extracts.

Production and certification of BOTS-1: bovine muscle-certified reference material for incurred veterinary drug residues.

A freeze-dried bovine muscle-certified reference material (CRM), known as BOTS-1 (DOI: https://doi.org/10.4224/crm.2018.bots-1 ), containing incurred residues of commonly used veterinary drugs was produced and certified for the mass fraction of eight veterinary drug residues. Value assignment was carried out using liquid chromatography tandem mass spectrometry (LC-MS/MS) methods in conjunction with isotope dilution and standard addition approaches involving stable isotope internal standards. Data from the National Research Council of Canada (NRC), Canadian Food Inspection Agency (CFIA), United States Department of Agriculture (USDA), and the Federal Office of Consumer Protection and Food Safety in Germany (BVL) were used for value assignment. Results for two drug residues were also obtained through an international inter-laboratory comparison CCQM-K141/P178 organized under the auspices of the International Bureau of Weights and Measures (BIPM). Quantitative NMR (1H-qNMR) was used to characterize primary standards of all veterinary drugs certified. The certified mass fractions of the veterinary drug residues were 490 ± 100µg/kg for chlorpromazine, 44 ± 4.4µg/kg for ciprofloxacin, 3.3 ± 1.4µg/kg for clenbuterol, 9.5 ± 0.8µg/kg for dexamethasone, 57 ± 4.8µg/kg for enrofloxacin, 3.0 ± 0.4µg/kg for meloxicam, 12.4 ± 1.2µg/kg for ractopamine, and 2290 ± 120µg/kg for sulfadiazine with expanded uncertainties quoted (95% confidence) which include the effects due to between-bottle inhomogeneity, instability during long-term storage and transportation, and characterization.

Gas Chromatography-Head Space-Mass Spectrometry Sensor based Quality Control of Dobutamine Hydrochloride Bulk Material for a mutagenic impurity, 2-bromopropane

In the process of making dobutamine hydrochloride, there is a possibility in the development of 2-bromopropane. The 2-bromopropane falls within the genotoxic group and is typically toxic and cancer suspicious agent. In order to monitor and then evaluate 2-bromopropane trace levels in dobutamine hydrochloride pharmaceutical samples, the gas chromatography technique with mass spectrometer analyser is described in this work. The separation of 2-bromopropane deployed DB-624 analysis column (30 m x 0.32 mm, 1.8 μm). The carrier gas utilized was helium flowing at a steady rate of 1.0 mL/min. At thermal setting of 220 °C, an injector operated in split mode (1:10). Selected ion monitoring pattern was choosen for the mass spectrometer analyzer to operate in. The applied technique provided adequate accuracy in addition to precision and superior linearity in the concentration span of 1.03 ppm to 15.00 ppm for 2- bromopropane. The approach was noted to be sensitive with LOD values for 2-bromopropane of 0.34 ppm. The recovery statistics of the standard addition approach provided affirmation that the stated method had efficaciously applied in terms of selectivity for enumeration of 2-bromopropane in dobutamine hydrochloride solutions with no interference.

Portable Electrochemical Platform With Carbon Fibre Microelectrodes Integrated on an OHP Sheet for Snake Venom Analysis.

Snake bite is a serious medical emergency often leading to untimely fatalities. Serotherapy is the only treatment method adapted for this, whose efficacy depends on identification of the Snake species and venom type. As a specific antivenom has to be implicated for saving the victim, in most of the cases, such identification is challenging, thus, leading to mortality due to delay in treatment or side effects of injecting polymeric non-specific antivenom. Therefore, a point-of-care, venom specific detection device could be an impactful diagnostic tool. Herein, a prototype of miniaturized electrochemical sensing platform is presented for detection of Crotaline, venom from various common pit viper snakes. A three electrode based micro-platform with carbon fibre microelectrode, modified with mesoporous carbon, embedded and laminated in commercial OHP sheet, has been developed. The dimensions of the miniaturized platform was 25 mm ×35 mm, size of electrode was 0.5 mm ×25 mm with an electrochemical testing zone of diameter 10 mm, electrode spacing as 3 mm. The microscopic characterization revealed immobilization of porous carbon on fine fibrous structure. The device gave highly stable and sensitive electro-catalytic oxidation of Crotaline at E'= at 0.81 V, and provided a linear range of 50- [Formula: see text], limit of detection as [Formula: see text] and limit of quantification as [Formula: see text]. The device exhibited negligible interference from physiological blood serum biochemicals, high stability and reproducibility. Further, real blood serum samples, analysis via standard addition approach, was performed which showcased appreciable recovery values confirming the practical applicability of the device.

A New Molecularly Imprinted Polymer for In-Tube SPME/UHPLC-MS/MS of Anandamide in Plasma Samples

Parkinson’s disease is a neurodegenerative disorder characterized by progressive loss of dopaminergic neurons. There is substantial evidence that the endocannabinoid system modulates the dopaminergic activity in the basal ganglia, a forebrain system that integrates cortical information to coordinate motor activity regulating signals. In this article, a fused-silica capillary with a molecularly imprinted polymer was developed for in-tube solid-phase microextraction of the endocannabinoid anandamide in plasma samples from Parkinson’s disease patients and further analysis by ultra-high-performance liquid chromatography with tandem mass spectrometry. The molecularly imprinted polymer capillary presented recognition sites with complementary shape, size, and functionality to anandamide. Scanning electron micrographs and Fouriertransform infrared spectra illustrated the physical and chemical modification of the printed and non-printed capillary surface. The in-tube solid-phase microextraction ultra-high-performance liquid chromatography with tandem mass spectrometry method presented a linear range from 0.1 to 20 ng mL-1, precision with coefficient of variation values ranging from 1.2 to 13%, and relative standard deviation accuracy ranging from -3.6 to 7.5%. The method developed herein can adequately determine anandamide in plasma samples from Parkinson’s disease patients. By applying the standard addition approach, the anandamide plasmatic concentration in these samples was found to range from 0.2 to 0.4 ng mL-1.

Are the (New) Synthetic Opioids U-47700, Tramadol and Their Main Metabolites Prone to Time-Dependent Postmortem Redistribution?-A Systematic Study Using an In Vivo Pig Model.

The interpretation of analytical results in forensic postmortem (PM) cases often poses a great challenge, in particular, due to possible PM redistribution (PMR) phenomena. In terms of new synthetic opioids, such data are usually not available and, if so, they are from case reports without the exact knowledge of dose, user habits, time of consumption or PM interval (PMI). Hence, a controlled toxicokinetic pig study was performed allowing the examination of PM tissue distribution and possible PMR of U-47700, tramadol and the main metabolites N-desmethyl-U-47700 and O-desmethyltramadol (ODT). For this purpose, 12 domestic pigs received an intravenous dose of 100 µg/kg body weight (BW) U-47700 or 1,000 µg/kg BW tramadol, respectively. The animals were put to death with T61 8 h after administration, and relevant organs, tissues and body fluids were sampled. Subsequently, the animals were stored at room temperature (RT), and the samples were taken again after 24, 48, and 72 h PM. Following homogenization and solid-phase extraction, quantification was performed applying a standard addition approach and liquid chromatography-tandem mass spectrometry. Only low-to-moderate concentration changes of U-47700, tramadol and their main metabolites were found in the analyzed tissue specimens and body fluids during storage at RT depending on the chosen PMI. On the contrary, a remarkable concentration increase of tramadol was observed in the liver tissue. These findings indicate that both synthetic opioids and their main metabolites are only slightly prone to PMR and central blood might be the matrix of choice for quantification of these substances.