Routine Laboratory Analysis Research Articles

NIR spectroscopy: Developing predictive models for chemical attributes and in vitro dry matter digestibility of Megathyrsus maximus cv. Tanzania

AbstractPastures in animal production are widely used, but production is conditioned to several factors. The nutritional composition of forage can be altered by soil conditions, season, plant maturity and morphology, so it is important to monitor its quality through chemical analysis. To optimize this type of analysis and speed up decision‐making by farmers and technicians, the use of near‐infrared spectroscopy (NIRS) is a tool that has been successfully applied. This research aimed to develop predictive models for chemical components of Megathyrsus maximus cv. Tanzania using NIR spectroscopy. Laboratory determinations of ash, crude protein (CP), in vitro dry matter digestibility (IVDMD), neutral detergent fiber (NDF) and acid detergent fiber (ADF) of 345 forage samples were used as reference data and correlated with their NIRS spectra. To calibrate the models, principal component analysis and partial least squares regression were applied. The results indicated that the prediction models of the studied parameters presented a coefficient of determination (R2) equal to or greater than 0.90; residual predictive deviation rate (RPD) greater than 3.0; error interval ratio (RER) greater than 12; close mean square error values between calibration and validation; and optimal number of latent variables (LV) between seven and eight for model calibration. For CP and IVDMD prediction, the regions with the highest simultaneous contribution were 1,414, 1996 and 2,384 nm; while for NDF and ADF, 1714, 1784, 1786, 2,160, 2,320 and 2,450 nm. The success in the development of predictive models by NIR spectroscopy to evaluate dry matter digestibility and other main chemical attributes of M. maximus cv. Tanzania shows that the quality of the models developed in this study enables them to be used alternatively in routine laboratory analysis in a quick, reliable and accurate way.

Sulfur Speciation in Li-S Batteries Determined by Operando Laboratory X-ray Emission Spectroscopy.

In this work, operando sulfur X-ray emission measurements on a Li-S battery cathode were performed using a laboratory setup as an alternative to more common synchrotron radiation based absorption studies. Photoexcitation by an X-ray tube was used. Valence-to-core Kβ X-ray emission spectra were recorded with a wavelength dispersive crystal spectrometer in von Hamos geometry, providing excellent energy resolution and good detection efficiency. The setup was used to record ex situ S Kβ emission spectra from S cathodes from the Li-S battery and also under operando conditions. Average S oxidation state within the battery cathode during battery cycling was determined from the shape of the Kβ emission spectra. A more detailed S species characterization was performed by fitting a linear combination of previously measured laboratory synthesized standards to the measured spectra. Relative amounts of different S species in the cathode were determined during the cycling of the Li-S battery. The main advantage of X-ray emission spectroscopy is that it can be performed on concentrated samples with S loading comparable to a real battery. The approach shows great promise for routine laboratory analysis of electrochemical processes in Li-S batteries and other sulfur-based systems under operando conditions.

Application of quality by design in the development of HPTLC Method for the estimation of ceritinib in bulk and synthetic mixture

The aim of the present work is to develop and validate the High-Performance Thin Layar Chromatography method for the estimation of Ceritinib in bulk and synthetic mixture using the Analytical Quality by Design (AQbD) approach as per ICH Q8 (R2). The AQbD is used to develop a precise HPTLC method. The AQbD parameter elements like QTMP (Quality Target Method Profile- Ceritinib, method type, instrument requirement, sample characteristics, synthetic mixture preparation) CMA—Critical method attributes (chloroform ± 1 ml, methanol ± 1 ml, triethylamine ± 0.05), and Risk assessment (the Ishikawa (fishbone) diagram) were carried out for the development of the HPTLC method for Cerinitib. The Box-Behnken design was utilized for the optimization of chromatographic conditions in HPTLC. The method utilized an aluminum HPTLC plate pre-coated with silica gel G60 F254 as a stationary phase along with chloroform: methanol: triethylamine (8.9: 1.6: 0.07 v/v/v) as a mobile phase. ANOVA results and perturbation plot indicates that methanol and triethylamine had more impact than chloroform on the retardation factor. The retardation factor was found to be 0.37. The absorbance mode at 277 nm was used for the densitometric study. It was found that the linearity ranged from 20 to 100 ng/band, with a correlation coefficient of 0.998. No interference was observed from excipients and mobile phase solvents. The results indicate that the AQbD approach is effectively applied to optimize the retardation factor and the method is appropriate for routine laboratory analysis.

Advanced HPTLC Method Development for Silibinin Analysis in Nanoformulated Scaffolds: A Box-Behnken Approach.

Silibinin (silybin), a bioactive component derived from the seeds of milk thistle (Silybum marianum), is recognized for its diverse pharmacological properties, including antioxidant, anti-inflammatory, and hepatoprotective effects. Given its therapeutic significance, accurately quantifying silybin in various formulations is essential. High-performance thin-layer chromatography (HPTLC) is a powerful analytical technique frequently used for this purpose. In this study, an HPTLC method was validated according to the International Council for Harmonization (ICH) guidelines to determine the concentration of silybin. The design of experiments (DoE), specifically the Box-Behnken design, was employed to optimize and understand the influence of critical method variables. The HPTLC method validation was performed using silica gel F254 HPTLC plates. The variables investigated included the composition of the mobile phase (% v/v), saturation time (minutes), and temperature in degree Celsius (°C), with the Box-Behnken design for optimization. The mobile phase consisted of chloroform, acetone, and formic acid in a 7:2:1 (v/v) ratio. Both the formulated scaffold and standard drug were applied to the plates, which were then processed in a twin chamber. After development, the plates were scanned at 288 nm using the Camag TLC Scanner IV with Vision CATS software. The validated HPTLC method demonstrated a strong linear relationship within the silybin concentration range of 2-10 μg/mL. The limit of detection (LOD) and limit of quantification (LOQ) for silybin were determined to be 0.469 and 1.423 μg/mL, respectively. Recovery studies indicated that the method provided accurate quantification, with recovery rates ranging from 97.53% to 99.82%. These results confirm the method's high accuracy, outstanding linearity, and reliability for the quantification of silybin in formulations. The validated HPTLC method proved to be a reliable analytical tool for the quantification of silybin in various formulations, particularly those containing polymers. The method's strong linearity, precision, and accuracy align with the ICH guidelines, making it suitable for routine analysis in quality control laboratories. The use of the Box-Behnken design for method optimization highlights the importance of systematic experimentation in achieving robust analytical outcomes.

Effect of Land Cover Variations on the Fertility Status of Soils of Umuhu in Ngor-Okpala Area of Imo State

The study was conducted to ascertain the effect of four different land covers (cassava cultivated field (CCF), oil palm plantation (OPP), plantain plantation (PP) and forest land (FL)) on the fertility status of soils of Umuhu in Ngor-Okpala area of Imo State, Nigeria. Random sampling technique was used in the collection of soil samples from the four different land covers. The sampling was done at the beginning, middle and end of each land cover. Twelve (12) composite samples were collected from all the land cover types with 3 per land cover. The composite samples were prepared for routine laboratory analyses. Data generated were analyzed statistically using analysis of variance. The textural classes of all the sampled soils were of loamy sand. The soil which had plantain plantation as its cover had the highest pH (6.4), Total N (0.229 %), Soil organic matter 4.70%, Available P (27.05 mg kg-1), Exch. Ca (6.492 cmolc kg-1), Exch. Mg (3.942 cmolc kg-1), Exch. K (0.953 cmolc kg-1), Exch. Na (0.06 cmolc kg-1), Exch. H (1.2 cmolc kg-1) and CEC (28.4 cmolc kg-1) with its Exch. Al in trace amount (<0.001 cmolc kg-1). The soil under CCF, despite being under low cover, did not show much degeneration due to the shifting cultivation being practiced in the study location. However, the result obtained from the study indicated that the different land covers especially that under plantain plantation (PP) have affected the soils fertility at different rates. This observed factor is strongly controlled by high content of organic materials, a dense vegetative cover which mitigates erosion effects and the addition of ash as a means of fertilizing the soils under plantain plantation being practiced in the study area.

The effects of drone transportation on routine laboratory, immunohematology, flow cytometry and molecular analyses.

Transportation of medical samples between laboratories or hospital sites is typically performed by motorized ground transport. Due to the increased traffic congestions in urban environments, drone transportation has become an attractive alternative for fast shipping of samples. In accordance with the CLSI guidelines and the ISO 15189 standard, the impact of this transportation type on sample integrity and performance of laboratory tests must be thoroughly validated. Blood samples from 36 healthy volunteers and bacterial spiked urine samples were subjected to a 20-40 min drone flight before they were analyzed and compared with their counterparts that stayed on the ground. Effects on stability of 30 routine biochemical and hematological parameters, immunohematology tests and flow cytometry and molecular tests were evaluated. No clinically relevant effects on blood group typing, flow cytometry lymphocyte subset testing and on the stability of the multicopy opacity-associated proteins (Opa) genes inbacterial DNA nor on the number of Abelson murine leukemia viral oncogene homolog 1 (abl) housekeeping genesin human peripheral blood cells were seen. Forthreeofthe 30 biochemistry and hematology parameters a statistically significant difference was found: gamma-glutamyl transferase (gamma-GT), mean corpuscular hemoglobin (MCH) and thrombocyte count. A clinically relevant effect however was only seen for potassium and lactate dehydrogenase (LDH). Multi-rotor drone transportation can be used for medical sample transportation with no effect on the majority of the tested parameters, including flow cytometry and molecular analyses, with the exception of a limited clinical impact on potassium and LDH.

Supplementation of vitamin E as an addition to a commercial renal diet does not prolong survival of cats with chronic kidney disease

BackgroundThe aim of this double-blind, placebo-controlled study was to investigate the effect of vitamin E supplementation as an addition to a commercial renal diet on survival time of cats with different stages of chronic kidney disease (CKD). In addition, we were interested whether vitamin E supplementation affects selected oxidative stress and clinical parameters. Thirty-four cats with CKD and 38 healthy cats were included in the study. Cats with CKD were classified according to the IRIS Guidelines; seven in IRIS stage 1, 15 in IRIS stage 2, five in IRIS stage 3 and seven in IRIS stage 4. Cats with CKD were treated according to IRIS Guidelines. Cats with CKD were randomly assigned to receive vitamin E (100 IU/cat/day) or placebo (mineral oil) for 24 weeks in addition to standard therapy. Plasma malondialdehyde (MDA) and protein carbonyl (PC) concentrations, DNA damage of peripheral lymphocytes and plasma vitamin E concentrations were measured at baseline and four, eight, 16 and 24 weeks thereafter. Routine laboratory analyses and assessment of clinical signs were performed at each visit.ResultsVitamin E supplementation had no effect on the survival time and did not reduce the severity of clinical signs. Before vitamin E supplementation, no significant differences in vitamin E, MDA and PC concentrations were found between healthy and CKD cats. However, plasma MDA concentration was statistically significantly higher (p = 0.043) in cats with early CKD (IRIS stages 1 and 2) than in cats with advanced CKD (IRIS stages 3 and 4). Additionally, DNA damage was statistically significantly higher in healthy cats (p ≤ 0.001) than in CKD cats. Plasma vitamin E concentrations increased statistically significantly in the vitamin E group compared to the placebo group four (p = 0.013) and eight (p = 0.017) weeks after the start of vitamin E supplementation. During the study and after 24 weeks of vitamin E supplementation, plasma MDA and PC concentrations and DNA damage remained similar to pre-supplementation levels in both the placebo and vitamin E groups.ConclusionsVitamin E supplementation as an addition to standard therapy does not prolong survival in feline CKD.

Advanced stability-indicating gradient elution reverse phase-high performance liquid chromatography method development and validation for simultaneous estimation of rutin and curcumin using an analytical quality by design approach

This research delves into a systematic Analytical Quality by Design (AQbD) approach to develop and validate a stability-indicating robust RP-HPLC method for the simultaneous determination of Rutin and Curcumin, compounds known for their anti-inflammatory, antioxidant, and wound-healing properties. Utilizing QbD principles, critical method parameters (CMPs) were identified and screened using design of experiments (DoE) and optimized via response surface methodology (RSM) with Central Composite Design (CCD). The optimized chromatographic separation used a Phenomenex Luna C18 column with gradient elution, using methanol and 0.1% formic acid, a flow rate of 1 ml/min, and detection at 345 nm (isosbestic point). The retention times were 5.63 mins for Rutin and 17.01 mins for Curcumin. The stability indication of the developed HPLC method was performed, and validation parameters were meticulously conducted following the ICH Q2(R1) guidelines, demonstrating linearity over 2-10 μg/ml for Rutin and 5-25 μg/ml for Curcumin, with correlation coefficients of 0.9999 and 0.9998, respectively. Precision studies showed %RSD within acceptable limits, and accuracies ranged between 97-100%. This validated HPLC method offers a reliable, sensitive approach for simultaneous quantifying Rutin and Curcumin, suitable for routine analysis in pharmaceutical laboratories and quality control.

Intravenous lipid emulsion interference in coagulation testing: an ex vivo analysis

Introduction Intravenous lipid emulsion is used in the rescue treatment of certain poisonings. A complication is interference with laboratory analyses. The aim of this study was to determine the impact of intravenous lipid emulsion on routine laboratory analysis of coagulation parameters ex vivo and determine if any of the analytical techniques remain reliable. Methods Samples were obtained from 19 healthy volunteers and divided in triplicate. One sample served as a control, and the other two were diluted to simulate the treatment of an average adult with Intralipid® 20 per cent Fresenius Kabi 100 mL (dilution-1) or 500 mL (dilution-2). Coagulation tests performed were prothrombin time, activated prothrombin time, D-dimer concentration and fibrinogen. Coagulation testing was performed by three techniques. Test-1 was performed on a Sysmex CN6000 analyzer. Test-2 was performed with a manual mechanical endpoint method using the semi-automated Stago KC4 Delta. Test-3 involved high-speed centrifugation before repeat testing on the Sysmex CN6000 analyzer. Results For test-1, only nine (47 per cent) samples in dilution-1 could be analyzed for coagulation tests, and no coagulation tests could be analyzed for dilution-2 because of lipaemia. For test-2 and test-3, all samples could be analyzed, and all results of both testing methods fell within the limits of the laboratory reference range. Discussion Difficulties in laboratory analysis of patients having received intravenous lipid emulsion are due to multiple factors. Most automated coagulation analyzers use optical measurements, which can be unreliable in the presence of a high intravenous lipid concentration. By altering the lipaemia in the testing solution using high-speed centrifugation or by using manual mechanical endpoint detection, we were able to obtain reliable results. These findings are limited by the use of an ex vivo method and healthy volunteers. Conclusions This ex vivo model confirms that Intralipid® interferes with routine coagulation studies. It is important that clinicians are aware and inform their laboratories of its administration.

Echocardiographic Changes in Prevalent Hemodialysis Population Based on Cardiac Symptomatology

BACKGROUND: Despite the prevalence of cardiovascular complications in hemodialysis (HD) patients, routine screening for asymptomatic individuals remains underexplored in clinical practice, particularly beyond those assessed for kidney transplantation. This study aimed to investigate echocardiographic changes in prevalent HD patients, irrespective of symptomatic presentation. SUBJECTS AND METHODS: A prospective, single-center study spanning 36 months included 79 HD patients. Grouping was based on cardiac symptomatology assessed with the New York Heart Association (NYHA) classification: group 1 comprised 18 asymptomatic patients (NYHA class I), while group 2 included 61 patients with moderate cardiac symptoms (NYHA classes II and III). Routine laboratory analyses, miRNA 133, hydration, and nutritional parameters were assessed, with echocardiography performed a day post-HD. RESULTS: Demographic data, HD quality, blood pressure, therapy, and most echocardiographic indicators showed no significant differences. Asymptomatic patients exhibited noteworthy cardiac structural and functional abnormalities: 55.5% had left ventricular hypertrophy (LVH), and 72.2% had increased left atrial volume index (LAVi). Concentric hypertrophy was observed in 7/18 asymptomatic patients, while 15/61 symptomatic patients demonstrated concentric left ventricle remodeling. In comparison to group 2, group 1 displayed higher hemoglobin, uric acid, and miRNA 133 concentrations, along with better hydration control and higher lean tissue index. All these dependent variables show statistically significant relation to the NYHA: approximately 13% of the variance in Hb levels and 18% of the variance in miRNA can be explained by NYHA classification. CONCLUSION: This study advocates for routine echocardiographic and cardiac examinations for all HD patients from the treatment's onset, irrespective of symptoms. Maintaining optimal volume, nutrition, urate concentrations, and hemoglobin is crucial for comprehensive cardiac care in this population. Additionally, exploring whether interventions targeting these non-cardiac factors could alleviate symptoms and improve quality of life in symptomatic HD patients would be worthwhile.

Mortality and COVID Infection: Predictors of Mortality 10 Months after Discharge.

The long-term survival of patients hospitalized with COVID-19 and the factors associated with poorer survival months after infection are not well understood. The aims of the present study were to analyze the overall mortality 10 months after admission. 762 patients with COVID-19 disease were included. Patients underwent a complete clinical evaluation, routine laboratory analysis and chest X-ray. Data collected included demographic and clinical data, such as vascular risk factors, tobacco or alcohol use, comorbidity, and institutionalization. Ten-month mortality was 25.6%: 108 deaths occurred in-hospital, while 87 patients died after discharge. In-hospital mortality was independently related to NT-proBNP values > 503.5 pg/mL [OR = 4.67 (2.38-9.20)], urea > 37 mg/dL [3.21 (1.86-7.31)] and age older than 71 years [OR = 1.93 (1.05-3.54)]. NT-proBNP values > 503.5 pg/mL [OR = 5.00 (3.06-8.19)], urea > 37 mg/dL [3.51 (1.97-6.27)], cognitive impairment [OR = 1.96 (1.30-2.95), cancer [OR = 2.23 (1.36-3.68), and leukocytes > 6330/mm3 [OR = 1.64 (1.08-2.50)], were independently associated with long-term mortality. the risk of death remains high even months after COVID-19 infection. Overall mortality of COVID-19 patients during 10 months after hospital discharge is nearly as high as that observed during hospital admission. Comorbidities such as cancer or cognitive impairment, organ dysfunction and inflammatory reaction are independent prognostic markers of long-term mortality.

Stability Indicating RP-HPLC Method Development and Validation for Imeglimin HCL in Pharmaceutical Dosage form

Background: Imeglimin HCL is a medication used in pharmaceutical products. To ensure its quality and effectiveness, it's crucial to have a reliable method to measure its concentration in these products. High-Performance Liquid Chromatography (HPLC) is a common technique for this purpose. Objective: This study aimed to develop and validate a precise and accurate stability indicating HPLC method for measuring Imeglimin HCL in pharmaceutical formulations. Methods: The method optimization process included selecting a suitable chromatographic column and determining the optimal mobile phase composition. A Credchrom C18 column (250mm x 4.6 mm x 5µm) was chosen, and a mobile phase consisting of Phosphate Buffer and acetonitrile (80:20, v/v) was identified as optimal. The selected conditions provided satisfactory resolution and a retention time of 2.5 minutes for Imeglimin HCL, with detection achieved at a wavelength of 241nm. Validation parameters, including linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ), were thoroughly evaluated. Precision was assessed through %RSD values for repeatability and intra-day and inter-day precision. Accuracy was determined by % recovery, with values ranging from 98.00% to 102%. Subsequently, force degradation studies were conducted to further assess the method's stability. Results: The developed RP-HPLC method exhibited excellent linearity (R^2 close to 1) and sensitivity, with LOD and LOQ values of 0.577722 μg/ml and 1.750673 μg/ml, respectively. Precision, as indicated by %RSD values, ranged from 0.994733 for repeatability to 0.988377–0.7480963 for intra-day precision and 0.988377–10.9883477 for inter-day precision. Accuracy, reflected by % recovery, was within acceptable limits (98.62–100.34%). These results, along with successful force degradation studies, confirmed the method's reliability and stability. Conclusions: In conclusion, the developed RP-HPLC method offers a sensitive, precise, and accurate means for the estimation of Imeglimin HCL in pharmaceutical formulations. Its robustness and stability make it suitable for routine analysis in pharmaceutical laboratories, ensuring the quality and efficacy of Imeglimin-containing products.

Analytical method development and validation for the simultaneous estimation of montelukast and Acebrophylline by RP- HPLC method

High performance liquid chromatography is at present one of the most sophisticated tool of the analysis. The estimation of Montelukast and Acebrophylline was done by RP-HPLC. The Phosphate buffer was pH 3.0 and the mobile phase was optimized with consists of Methanol: Phosphate buffer mixed in the ratio of 70:30 % v/ v. Inertsil C18 column C18 (4.6 x 150mm, 5mm) or equivalent chemically bonded to porous silica particles was used as stationary phase. The detection was carried out using UV detector at 260 nm. The solutions were chromatographed at a constant flow rate of 0.8 ml/min. the linearity range of Montelukast and Acebrophylline were found to be from 100-500 mg/ml of Montelukast and 1-5mg/ml of Acebrophylline . Linear regression coefficient was not more than 0.999.The values of % RSD are less than 2% indicating accuracy and precision of the method. The percentage recovery varies from 98-102% of Montelukast and acebrophylline. LOD and LOQ were found to be within limit. The results obtained on the validation parameters met ICH and USP requirements .it inferred the method found to be simple, accurate, precise and linear. The method was found to be having suitable application in routine laboratory analysis with high degree of accuracy and precision.

A comparative analysis of univariate versus multivariate eco-friendly spectrophotometric manipulations for resolving severely overlapped spectra of vonoprazan and amoxicillin new combination

Vonoprazan and amoxicillin are pharmacological combinations that demonstrate synergistic effects in treating Helicobacter pylori (H. pylori), a global public health concern associated with peptic ulcer disease and gastric cancer. Four spectrophotometric methods were developed, including two univariate techniques (Fourier self-deconvolution and ratio difference) and two multivariate chemometric approaches (partial least squares and principal component regression). These methods provide innovative solutions for effectively resolving and accurately quantifying the overlapping spectra of vonoprazan and amoxicillin. The concentration ranges covered were 3–60 μg ml−1 for vonoprazan and 5–140 μg ml−1 for amoxicillin. To assess the environmental sustainability of the methodologies, various measures such as the Green Analytical Procedure Index (GAPI), National Environmental Method Index (NEMI), Analytical GREEnness Calculator, and Analytical Eco-scale, as well as RGB12 and hexagon toll were implemented. The validation of the developed techniques was carried out in compliance with ICH standards. The present study is highly significant because it is the first time that the mixture has been determined using the current approaches. The comparative analysis demonstrated no significant difference in terms of accuracy and precision compared to reference HPLC method (p = 0.05). The established spectrophotometric methods offer a straightforward, rapid, and cost-effective alternative to complex analytical techniques for determining the vonoprazan and amoxicillin mixture. They show potential for routine analysis in research laboratories and pharmaceutical industries.

Two green and sensitive spectrofluorimetric approaches for determination of Ambroxol and guaifenesin in their single and combined pharmaceutical formulations.

Ambroxol hydrochloride (AMX) and guaifenesin (GFN) are approved drugs utilized to treat coughs through their potent mucolytic and expectorant properties. Due to their massive, combined administration in many illnesses, there is a persistent need for their concurrent estimation in different pharmaceutical formulations. Two sensitive, environmentally friendly spectrofluorimetric methods were developed. AMX was determined using the first method (I) without interference from GFN. This method depends on the quenching of Erythrosine B (EB) native fluorescence at 552 nm after excitation at 527 nm due to the formation of a non-fluorescent AMX-EB ion-pair complex in Britton-Robinson buffer (BRB) solution pH (3.5). The concentration plot is linear over the 0.25-5.0μg/mL range, with a mean percent found value of 99.74%. Method (II) depends on measuring the native fluorescence of aqueous GFN solution at two analytical wavelengths, either 300 or 600 nm, after excitation at 274 nm. Relative fluorescence intensity (RFI)-concentration plots are linear over the ranges of 0.02-0.5 and 0.1-2.0μg/ml, with mean percent found at 99.96% and 99.91% at dual wavelengths, respectively. The proposed methods were successfully applied to assay both drugs in raw materials and different single and combined pharmaceutical formulations. These methods have been thoroughly validated following International Committee on Harmonisation (ICH) guidelines. National Environmental Methods Index, Analytical Eco-Scale, and Green Analytical Procedure Index were used to prove greenness, thereby enhancing their applicability. The proposed techniques provide straightforward, precise, and cost-effective solutions for routine formulation analysis in quality control laboratories.

Analytical Method Development and Validation: Quantitative Estimation for Assay of Revefenacin in Pharmaceutical Dosage form by RP-HPLC method

Revefenacin is a medication for the treatment of chronic obstructive pulmonary disease (COPD). A simple, accurate and precise reversed-phase high-performance-liquid-chromatography (RP-HPLC) method was developed and validated for Revefenacin in bulk and pharmaceutical dosage forms. The method was optimized by selecting chromatographic conditions of 30:70 v/v, 0.2% Perchloric acid solution: Acetonitrile used as a mobile-phase, Inertsil® column ODS-3V (4.6mm* 25 cm, 5μm), injection volume 20μL, flow-rate 1.0ml/min at an ambient temperature of column oven (25°C), auto-sampler temperature 15°C and λmax 242nm. A different column of C18-Kinetex F-5 (4.6mm* 25 cm, 5μm) was also examined, reporting no significant divergence in the results obtained. The method was further validated, giving good precision (%RSD/CV<1.0) and acceptable linearity (r≥0.999) on both columns. The pharmaceutical revefenacin drug offers a high recovery rate of 99.77%. The proposed method is observed as simple, rapid and economical and can be employed for routine analysis in quality control laboratories.

Characterization of ready-mixed plasters for the conservation of architectural heritage

Nowadays, the use of ready-mixed mortars is widespread in the choice of materials for the conservation of architectural heritage. These are dry mixtures, which contain binders, aggregates and additives and which require only the addition and mixing with water. Their use makes it possible to avoid errors in the ratios of the components, limits waste and guarantee a constant product quality. On the other hand, there are no standards that require the declaration of all the components in the mixture, which makes difficult to select the most appropriate materials to use on historical substrates. Indeed, the presence of certain additives could alter other properties of the mortars or increase the probability of developing forms of degradation. In addition, the mixtures on the market are frequently labelled as environmentally, human and animal health friendly; but the actual sustainability of these mixtures is difficult to ascertain, because there are still few companies that study the Life Cycle Assessment (LCA) and publish the Environmental Product Declaration (EPD) of their products. In order to deepen the knowledge of this class of materials, this study proposes the characterization of five ready-mixed plasters, all labelled green by the companies. Analyses (X-ray diffraction, infrared spectroscopy, thermal analysis) have been carried out firstly to assess the compatibility of the mixtures with historical materials, to compare the results obtained with what is stated by the companies in the product data sheets, and to assess whether, through routine laboratory analyses, it is possible to certify the sustainability of these materials.

Green and High Throughput Assay Using 96-Microwell Base to Determine Metformin Hydrochloride in the Tablet Dosage Form.

Metformin (MET) is an oral antidiabetic drug widely used as the primary treatment for type 2 diabetes mellitus (T2DM). While various spectrophotometric assays exist for determining MET in pharmaceutical formulations, they often have limited throughput for quality control purposes. This study describes the validation of a 96-microwell plate spectrophotometer method using charge-transfer complexes (CTCs) with chloranilic acid (CLA) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) for the quality control and detected of MET. This reaction was carried out in 96-microwell plates, and the absorbance of the colored complexes of CLA and DDQ were measured at 530 nm and 460 nm, respectively, using an absorbance microplate reader. This study aims to identify and quantify the use of a 96-microwell plate spectrophotometer analytical technique for assessing complicated formulations. The method was successfully used for the quantification of MET in the tablet dosage form. The results showed good correlation coefficients (0.996 and 0.997) with CLA and DDQ, respectively. The present method showed high precision with RSD % not exceeding 2.17%. The accuracy of the method was obtained by recovery percentage, with percentage values less than ±5%. The Analytical Greenness Metric (AGREE) was used to evaluate greenness of the assays. The result show that the microwell assay method is greenness and suitable for handling large samples on a daily used with high throughput analysis. The use of the 96-microwell-plate method is superior to the existing method in terms of simplicity of the procedure, the low economic cost, and its consumption of low amounts of reagents and organic ethanol solvent, making it an environmentally friendly method. Therefore, these advantages make them suitable and rapid alternatives method to current methods for routine metformin analysis in quality control laboratories.

O impacto da apresentação clínica, presença de SIRS e disfunção de órgãos em cadelas com piometra

ABSTRACT: This retrospective study evaluated the impact of the clinical presentation of pyometra on the morbidity and mortality of the affected bitches. The medical records of 47 bitches surgically treated for pyometra between May and December 2017 were evaluated. The admission data were analyzed to classify the animals according to the clinical presentation of pyometra, presence of systemic inflammatory response syndrome (SIRS) and organic dysfunctions (cardiovascular, neurological, renal, and hemostatic). The outcomes were recorded for up to 14 days. The overall mortality rate was 13%. Open and closed pyometra were identified in 62% (n= 29) and 38% (n= 18) of the cases, respectively. The SIRS was present in 44 (94%) bitches, and 27 (57%) had at least one organ dysfunction. The most prevalent dysfunction was hemostatic, followed by neurological, cardiovascular, and renal dysfunction. The clinical presentation of pyometra and the presence of SIRS did not correlate with each other, or with the presence of organ dysfunction and mortality. However, cumulative organ dysfunction (r=0.532, P=<0.001) and the presence of neurological (r= -0.371; P=0.012) and renal dysfunction (r=0.303; P=0.041) were correlated with mortality. Thus, the identification of organ dysfunction through physical examination and routine laboratory analysis proved to be more effective in identifying and predicting the critically ill patients than the SIRS criteria or clinical presentation of the disease.

Development and validation of a test system for determining the polymorphism in the DGKH and PPP1R1C genes associated with body weight of sheep

Relevance. National sheep breeding is lagging behind other branches of animal husbandry considering using of modern DNA technologies. The search for new genes — potential candidates associated with economically significant traits — is relevant for a more complete disclosure of the genetic potential of sheep. An earlier genome-wide association study showed that the DGKH and PPP1R1C genes had a certain effect on body weight of sheep at birth and at the age of 90 days. In this regard, a more detailed study of polymorphism in the DGKH and PPP1R1C genes can deepen understanding of growth and development processes in domestic sheep, so we have chosen these genes as targets for the experiment.Methods. Primers and probes were selected for amplification of a fragment with targeted SNPs in the DGKH and PPP1R1C genes with a length of 68 nucleotide pairs based on the reference DNA sequence on the 10th (NC_056063.1) and the 2nd chromosome (NC_056055.1) sheep represented in NCBI. To determine polymorphism, real-time PCR-based test systems were developed. Genotypes were determined using a multiparametric graph. The test systems were tested on 147 sheep of the southern meat breed.Results. The developed test systems for promising DGKH and PPP1R1C genes allowed to clearly determine sheep genotypes based on using the PCR-RT. All three genotypes (T/T, C/T, and C/C) were identified in the DGKH gene. No homozygous genotype (T/T) was identified in the PPP1R1C gene in the studied population. The test systems are suitable for routine DNA analysis in molecular genetic laboratories.