Precipitation Step Research Articles

SERS-Driven Ceftriaxone Detection in Blood Plasma: A Protein Precipitation Approach

Accurate detection of antibiotics in biological samples is essential for clinical diagnoses and therapeutic drug monitoring. This research examines how proteins and other substances in blood plasma affect the detection of the antibiotic ceftriaxone using surface-enhanced Raman spectroscopy (SERS). We detected ceftriaxone spiked in blood plasma without sample preparation within the range of 1 mg/mL to 50 µg/mL. By employing a pretreatment approach involving methanol-based protein precipitation to eliminate interfering substances from a spiked blood plasma solution, we could detect ceftriaxone down to 20 µg/mL. The comparative analysis demonstrates that the protein precipitation step enhances the sensitivity of SERS-based detection of drugs in the matrix blood plasma. The insights derived from this study are highly beneficial and can prove advantageous in developing new antibiotic detection methods that are both sensitive and selective in complex biological matrices. These methods can have important implications for clinical treatments.

B-174 Quantification of vitamin C in serum as a biomarker using the Laser Diode Thermal Desorption technique “LDTD”/Mass Spectrometry (Luxon)

Abstract Background Ascorbic acid (AA) or vitamin C, is a crucial substance that is involved in numerous physiological processes in living beings, including collagen production and antioxidant activity. When ascorbic acid is exposed to moisture, air, heat, light and oxygen, it oxidizes and turns into dehydroascorbic acid (DHAA). Our objective is to describe the sample preparation procedure and total quantification of AA and DHAA in human serum sample based on LDTD-MS/MS, and demonstrate its efficiency in quantification, combined with analysis selectivity and rapidity. Methods The Luxon is a sample introduction and ionization source that used a fiber-coupled laser diode to thermally desorb neutral molecules and charge them using a corona needle so they can be analyzed by mass spectrometry. Due to the instability of ascorbic acid, plasma samples were enriched and stabilized with an acidic solution. Sample preparation utilized a phosphate-containing buffer to minimize interferents and help volatize the analyte, followed by a precipitation step using acetonitrile to further clean the extract. Stable isotope-labeled internal standard (13C6 ascorbic acid) and homemade calibration curve and water quality controls were used for quantification. The analysis was performed using a Sciex 6500+ Triple Quad in APCI (Atmospheric Pressure Chemical Ionization) negative mode. Calibration curves ranging from 1 to 100 μg/mL and QCs were prepared in ascorbic acid-depleted serum (kept at room temperature/exposed to light for 4 days). The peak area against the internal standard (IS) ratio was used to normalize the signal. The calibration curves were plotted using the peak area ratio and the nominal concentration of standards. The following parameters were assessed for quantitative analysis: linearity, precision, accuracy, carryover and parallelism. A linear regression model with 1/X2 weighting was used to obtain the calibration curve equation and determine the correlation coefficient (r2). Parallelism was assessed by comparing the correlation coefficient of calibration curves prepared in water to six curves prepared with biological matrixes. Intra and inter assay precision and accuracy were determined by measuring QC samples ranging from low to high concentration in five replicates for 3 consecutive days. Results Calibration curves (n=3) showed excellent linearity. The calibration curve resulted in a linear response throughout the concentration range, the correlation coefficient was 0.99. For each quality control concentrations CVs were <10% complying with desirable specifications for imprecision and accuracy. Intra-assay CVs were 3.0-8.89% and inter-assay CVs were 7.64-9.79% for precision which reflects the agreement between repeated measurements. The results for accuracy showed similar results, intra-assay CVs were 3.83-9.20% and inter-assay CVs were 5.0-9.7%. Conclusions The new LDTD-MS/MS method described here has been validated as an effective technique to reliably and rapidly quantify ascorbic acid. Simple sample preparation, rapid acid stabilization combined with high yield allow analysis without degradation responses.

Synergistic effects of Paenibacillus mucilaginosus and Penicillium pimiteouiense on the extraction of humic substances from lignite

Based on the common step of alkaline extraction-acid precipitation, lignite treatment techniques have been developed to improve extraction efficiency and yield of humic acid (HA). This study explored the combination of bacterium Paenibacillus mucilaginosus (P.m) LT1906 and fungus Penicillium pimiteouiense (P.p) LL2205 strains for pre- and post-treatment of lignite collected from eastern Inner Mongol to enhance HA and fulvic acid (FA) yield and plant growth-promoting activity. The lignite was subjected to P.m followed by HNO3 treatment, HNO3 followed by P.p treatment, and a combination of P.m, HNO3 and P.p treatments, then performed by alkaline extraction-acid precipitation technique. The results indicated that P.m pretreatment increased HA yield by 12.47 %, while P.p posttreatment significantly enhanced FA yield by 13.37 %. Combined bacterial and fungal treatments notably improved the total yields of HA and FA by 19.12 %. Structural analysis showed that P.m pretreatment caused the deformation on the lignite surface and the complete lignite oxidation and leaching reactions, thereby increasing the production of HA, yet had no impact on the elemental composition or chemical structure of HA. P.p posttreatment introduced oxygencontaining functional groups and reduced aromatic components through oxidative depolymerization of lignite, leading to increased FA production and HA seed germination-promoting activity.

Ocular pharmacokinetics of acetazolamide from intravitreal implants by high-performance liquid chromatography coupled to tandem mass spectrometry

Glaucoma, a leading cause of irreversible blindness, affects about 70 million people globally. Its treatment focuses on reducing intraocular pressure. Acetazolamide, a potent anti-glaucoma drug, is currently used only systemically due to low solubility and permeation, which cause severe side effects. Developing topical medications with acetazolamide requires robust analytical methods for its detection in biological samples. In this context, this study aimed to develop a method to quantify acetazolamide in rabbit vitreous humor samples. The method involved a simple, fast, inexpensive, and environmentally friendly protein precipitation step for sample preparation, needing just 50 μL of sample and 200 μL of organic solvent, with adequate recovery. This was combined with high-performance liquid chromatography coupled to tandem mass spectrometry, enabling highly sensitive (LOQ of 5 ng/mL) quantification within only 5 min. The method proved to be selective, precise, and accurate, with well-fitted analytical curves, with no carryover, and no matrix effect impacting reliability. The method was successfully applied to analyze vitreous humor samples from rabbits in pharmacokinetic studies, monitoring drug release from intravitreal implants. Results showed a controlled release profile, with a maximum drug concentration (Cmax) of 426.01 ± 64.57 ng/mL, time to reach Cmax (Tmax) of 28 days, and area under the curve (AUC0–42 and AUC0-∞) of 7722.66 ± 1125.96 ng days/mL and 8998.11 ± 1311.92 ng days/mL, respectively. The device demonstrated significantly slower elimination, ensuring therapeutic levels for an extended period when compared to intravitreal injection.

Design and synthesis of high quantum yield doped carbon nano probe derived from household sources for sensing of the anti-GERD drug pantoprazole

This contribution aims to design and validate a new green, cheap, and fast approach for determining the anti-GERD drug pantoprazole in different matrices. New S and N-doped carbon nanomaterials (S,N-CNMs) have been prepared via microwave irradiation of a mixture of widely available household sources. Remarkably, the utilization of a blend of carbamide and thiocarbamide with table sugar yields S,N-CNMs exhibiting the utmost quantum yield (54 %), hydrophilicity, as well as stable, homogeneous, and diminutive particle size distribution. Fourier transform infrared spectroscopy, transmission electron microscopy, spectrophotometry, and fluorescence spectroscopy were applied to characterize the S,N-CNMs. The S,N-CNMs have been used as a turn-off fluorescence probe to determine pantoprazole via a synergism of the inner filter effect and static quenching mechanisms. The fluorescence quenching is linearly correlated to pantoprazole concentration over the range of 1.0–25.0 µg/mL with a detection limit of 0.16 µg/mL. The developed probe exhibited good selectivity for pantoprazole in the presence of variability of substances. Therefore, it was applied for quality control of pantoprazole in pharmaceutical tablets and vials with an average recovery % of 100.10 ± 0.77 % and 100.33 ± 0.92 %, respectively. Moreover, it was successfully implemented to examine the content uniformity of pantoprazole in tablets. Furthermore, the prepared S,N-CNMs have been successfully used for the analysis of pantoprazole in human plasma after a simple protein precipitation step with a recovery % of 97.88 ± 5.72 %. The greenness and blueness of the developed method have been positively assessed by recent tools showing the eco-friendliness and applicability of the developed method.

Comparative Analysis of Digestion Methods for Bile Proteomics: The Key to Unlocking Biliary Biomarker Potential.

Bile's potential to reflect the health of the biliary system has led to increased attention, with proteomic analysis offering deeper understanding of biliary diseases and potential biomarkers. With the emergence of normothermic machine perfusion (NMP), bile can be easily collected and analyzed. However, the composition of bile can make the application of proteomics challenging. This study systematically evaluated various trypsin digestion methods to optimize proteomics of bile from human NMP livers. Bile was collected from 12 human donor livers that were accepted for transplantation after the NMP viability assessment. We performed tryptic digestion using six different methods: in-gel, in-solution, S-Trap, SMART, EasyPep, and filter-aided sample purification, with or without additional precipitation before digestion. Proteins were analyzed using untargeted proteomics. Methods were assessed for total protein IDs, variation, and protein characteristics to determine the most optimal method. Methods involving precipitation surpassed crude methods in protein identifications (4500 vs 3815) except for in-gel digestion. Filtered data (40%) resulted in 3192 versus 2469 for precipitated and crude methods, respectively. We found minimal differences in mass, cellular components, or hydrophobicity of proteins between methods. Intermethod variability was notably diverse, with in-gel, in-solution, and EasyPep outperforming others. Age-related biological comparisons revealed upregulation of metabolic-related processes in younger donors and immune response and cell cycle-related processes in older donors. Variability between methods emphasizes the importance of cross-validation across multiple analytical approaches to ensure robust analysis. We recommend the in-gel crude method for its simplicity and efficiency, avoiding additional precipitation steps. Sample processing speed, cost, cleanliness, and reproducibility should be considered when a digestion method is selected for bile proteomics.

Selective Leaching and Separation of Uranium from Ochre-Umm Greifat, Red Sea Coast, Central Eastern Desert, Egypt

The Ochre-Umm Greifat area is one of the Red Sea areas with high concentrations of iron and zinc, which is formed from hydrothermal solutions as a result of the structural activity that occurred in the Red Sea Zone during the Pleistocene period. These deposits are also accompanied by deposits of low- to high uranium grade. In addition to Zn, Pb, and Cu anomalies, particularly in fault zones and their branches affecting the study area, although there are numerous zinc minerals in the Ocher-Greifat area, uranium minerals are scarce, with only one mineral, compreignacite, being recorded and the majority of the uranium being present as an adsorbed element on iron and/or clay stones. In addition, uranothorite is extremely rare and occurs as fine grains embedded in rocks. A technological sample was taken from an iron-rich clay area in a fault zone and was found to assay 700-ppm uranium. The leachability of uranium from the used sample was investigated using an alkaline solution based on the chemical and mineralogical composition of the used sample. The selected ore is treated with Na2CO3 and NaHCO3 in the presence of H2O2 as oxidant. Many digestion factors are studied and optimized. Under the optimum leaching conditions, the uranium dissolution efficiency is around 84%. For the uranium separation, the pH of the leach liquor is adjusted at 10, then subjected to a solvent extraction step using 4% Aliquat®336/kerosene in the presence of isodecanol as third-phase prevention. The loaded organic solvent was then treated with NaOH/H2O2 solution as a stripping solution. Finally, the resultant solution is subjected to a precipitation step using ammonia solution.

A process to utilize lithium and magnesium resources from low-magnesium salt lake brine through extraction and CO2 stripping

The lithium resources in Tibet's salt lake account for more than 31 % of China's total, featuring a low magnesium-to-lithium ratio with its carbonate-type salt lakes representing a unique resource of the region. When extracting lithium from low-magnesium salt lakes (carbonate-type salt lakes) using a β-diketone system, it is common practice to add alkaline reagents at the front end to precipitate and remove magnesium. This study utilizes the LIX54/TRPO-ADD-1 system for the co-extraction of lithium and magnesium from the brine of the Jiezechaka salt lake, employing CO2 for stepwise stripping to separate lithium and magnesium, using a weak acid instead of a strong acid for stripping to avoid impacting Tibet's fragile ecological environment. By controlling the pH with varying amounts of CO2, an extraction efficiency of 92.54 % for lithium at pH=8.5 and an A/O = 1:5, and complete stripping of magnesium from the loaded organic phase at pH=7 with an A/O = 1:3.2 over three theoretical stages was achieved. This new process omits the front-end magnesium precipitation step, offering a new avenue for the separation of lithium and magnesium in salt lakes and the resource utilization of magnesium.

Modulation of physico-chemical and technofunctional properties of quinoa protein isolate: Effect of precipitation acid

The typically low solubility and gelation capacity of plant proteins can impose challenges in the design of high-quality plant-based foods. The acid used during the precipitation step of plant protein isolate extraction can influence protein functionality. Here, acetic acid and citric acid were used to extract quinoa protein isolate (QPI) from quinoa flour, as these acids are more kosmotropic than the commonly used HCl, promoting the stabilisation of the native protein structure. While proximate analysis showed that total protein was similar for the three isolates, precipitation with kosmotropic acids increased soluble protein, which correlated positively with gel strength. Microstructure analysis revealed that these gels contained a less porous protein network with lipid droplet inclusions. This study shows that the choice of precipitation acid offers an opportunity to tailor the properties of quinoa protein isolate for application, a strategy that is likely applicable to other plant protein isolates.

NdFeB Magnets Recycling via High-Pressure Selective Leaching and the Impurities Behaviors

Global concerns about climate change are driving increased demand of electric vehicles for sustainable transportation and turbines in emerging energy solutions, where permanent magnets (PMs) and rare earth elements (REEs) play a critical role. However, global REEs recycling rates are only 3% and 8% for light and heavy REEs, respectively. This work proposes an effective approach to separate the REEs and iron via high-pressure selective leaching by low-concentrated nitric acid from the end-of-life NdFeB magnet and investigates the impurities behavior during the leaching and precipitation steps. The results from the optimized leaching conditions demonstrated over 95% REEs leaching efficiency with less than 0.3% Fe dissolution. Approximately 70% of Al and B were leached as well, while other elements (Co, Ni, Cu) had leaching efficiencies below 40%, leaving a hematite rich residue. Adjusting the pH removes Al and Fe in leachate but minimally affects Cu, Co, and Ni. Na2S addition is more effective against transition metals, but both methods result in around 10% REEs loss. Direct oxalate precipitation is suggested for the obtained leachate, which can yield over 97.5% REEs oxides with approximately 1.0% alumina, which is acceptable for magnet remanufacturing due to the aluminum content commonly found in magnets. The technology developed in this study offers opportunities for closed-loop recycling and remanufacturing of PMs, benefiting the environment, economy, and supply chain security.Graphical

Upcycling spent Ni–Cd battery for selective synthesis of nanoparticles to harness into electro-conductive thin films: An eco-friendly approach for e-waste management

Inspired by Sustainable Development Goal-12 (responsible consumption and production) to reduce e-waste, an environment friendly extraction of Cd, Ni and Co from Ni–Cd spent batteries have been reported here using ammonium chloride as a chlorinating agent for the first time. Multiple parameters i.e. dose of chlorinating agent, roasting time, and temperature has been optimized that resulted in extraction of 99.93 % Cd, 90.1 % Ni and 84.81 % Co. Metal enriched leachate was then subjected to unique precipitation steps that resulted in synthesis of Cd nanoparticles, Co3O4·NiO composite nanoparticles, and NiO nanoparticles, characterized through XRD and FESEM-EDX. Synthesized nanoparticles were further employed in the fabrication of electro-conductive nano thin films via Layer by Layer (LbL) technique. Film buildup of six novel architectures was monitored using an Ellipsometer. Homogenous film growth was tailored by optimizing ratio, concentration and dipping time of nanoparticles and polymer solutions. Conductivity studies revealed that imbedding synthesized nanoparticles impart significant conductive behavior in thin films with maximum conductivity of 1.8 × 10−4 S/cm, in addition to exhibiting appreciable resistance against 0.1 M HCl, 0.1 M NaOH and acetone. Contact angle and AFM images revealed that the films were slightly wettable and homogenous respectively. Conclusively not only Cd, Ni and Co were successfully recycled from Ni–Cd spent batteries employing greener technique but were also harnessed into robust nano-thin films with focus towards electro-conductivity based applications.

Process Design for Direct Production of Battery Grade Nickel Sulfate

The clean energy transition has increased the global demand of nickel sulfate used in the Li-ion batteries. A short-term solution is to refine the nickel sulfate product from nickel intermediates. In the long-term, new direct nickel sulfate production technologies are needed. This research focused on the modeling-based concept development of a novel direct hydrometallurgical nickel sulfate process consisting of chemical leaching, impurity removal by precipitation, solvent extraction, and crystallization as an alternative to the conventional nickel sulfate production route via a nickel matte intermediate. The conventional process route with the studied nickel concentrate had lower chemical consumption and waste production compared to direct hydrometallurgical process where approximately 60% of iron was leached consuming oxygen, and the following iron precipitation step consuming calcium carbonate resulted in a high amount of iron precipitate together with gypsum. However, hydrometallurgical alternatives are often suitable for lower ore grades or volumes and can recover copper as by-product metal. The biggest impacts on carbon footprint from chemical consumption in the direct hydrometallurgical process were generated in iron precipitation and oxygen use in leaching. With the studied nickel concentrate, pyrrhotite played a key role in both oxygen use and iron precipitation. In the leaching step, 68% of total oxygen consumption was related to pyrrhotite leaching, while in iron removal 73% of total iron originated from pyrrhotite. Thus, especially pyrrhotite removal prior to leaching needs to be developed to reduce the carbon dioxide footprint, when the pyrrhotite content in the material is high.Graphical

Isolation and purification of a monoclonal antibody from a cell culture supernatant by multistage precipitation and solid-liquid extraction

A process of multistage pH mediated precipitation was developed for capture and purification of a monoclonal antibody (mAb) from a Chinese hamster ovary cell culture (CHO) harvest. The mAb was separated from low molecular mass impurities (LMWI) and high molecular weight impurities (HMWI) by a sequence of two precipitation steps or by a sequence of precipitation and solid-liquid extraction (SLE). In the former approach, HMWI were precipitated from the harvest at pH 5 in the presence of polyethylene glycol (PEG), whereas the obtained supernatant was subsequently subjected to precipitation at pH 8 to obtain the mAb of 96 % chromatographic purity in a precipitate form. In the latter approach, the mAb was precipitated from the harvest at pH 8 in the presence of PEG to isolate the mAb from LMWI, then the precipitate obtained was subjected to SLE to extract the mAb of 99 % purity in a form of liquid solution. The coupling of precipitation at pH 8 and SLE allowed effective removal of DNA from 9260 ng/mL in the feed to 1.4 ng/mL in the product of two-stage SLE, and host cell proteins (HCP) from 300 µg/mL to 0.07 µg/mL, which was not possible by the coupling of two precipitation stages at pH 5 and 8. Therefore, the combination of precipitation and SLE can be a promising technique for capture of mAbs from CHO cell harvests.

A simple method for the determination of acyclovir concentrations in human plasma using high-performance liquid chromatography

BackgroundAcyclovir is an anti-viral medication given to treat herpes simplex and herpes zoster infection. In some severe conditions such as herpes encephalitis, acyclovir is administered intravenously. However, high acyclovir doses may cause acute kidney injury and low acyclovir dose may predispose the patient to inadequate exposure to acyclovir which could be fatal in some conditions. In such cases, the acyclovir plasma concentrations will potentially guide the diagnosis and management of the kidney injury. In this study, we provide a simple and time-efficient method for analyzing acyclovir in human plasma using high-performance liquid chromatography (HPLC).ResultsThe process starts with a single protein precipitation step by adding acetonitrile to deproteinize 300 µL of plasma. The chromatographic separation conditions consist of a mobile phase of water: methanol (97:3, v/v), a flow rate of 1 mL/min, a run time of 17 min, and a detection wavelength of 254 nm. The calibration curve was linear over the range of (0.70–60 mg/L) (r2 ˃ 0.99). The retention times of acyclovir and the internal standard were around 15 and 12 min, respectively. The intra-day and inter-day analysis of acyclovir in plasma using this method exhibited accuracy and precision of less than 7%, which lies within the acceptable range. Different greenness assessment tools confirmed that the proposed method is eco-friendly.ConclusionThe proposed method of analysis of acyclovir in the plasma using HPLC is simple, green and accurate method. This method could be applied in clinical settings where monitoring acyclovir concentrations is essential as it has wide range of the concentrations that could be detected.

Development and Validation of a Simple and Reliable HPLC-UV Method for Determining Gemcitabine Levels: Application in Pharmacokinetic Analysis.

Background and Objectives: Gemcitabine has been used to treat various solid cancers, including, since 1997, metastatic pancreatic cancer. Here, we developed an HPLC-UV method to determine serum gemcitabine levels and use it in pharmacokinetic studies. Materials and Methods: The analysis was performed after a single protein precipitation step on a reversed-phase column, isocratically eluted with sodium phosphate buffer and methanol. For the pharmacokinetic study, NOD/SCID mice received a single dose of gemcitabine at 100 mg/kg by either subcutaneous (SC) or intraperitoneal (IP) administration. Blood samples were collected at 5, 15, and 30 min and 1, 2, 4, and 6 h after the administration of gemcitabine for further analysis. Results: The duration of the analysis was ~12.5 min. The calibration curve was linear (r2 = 0.999) over the range of 1-400 μM. The mean recovery of GEM was 96.53% and the limit of detection was 0.166 μΜ. T1/2, Tmax, Cmax, AUC0-t, and clearance were 64.49 min, 5.00 min, 264.88 μmol/L, 9351.95 μmol/L*min, and 0.0103(mg)/(μmol/L)/min, respectively, for the SC administration. The corresponding values for the IP administration were 59.34 min, 5.00 min, 300.73 μmol/L, 8981.35 μmol/L*min and 0.0108(mg)/(μmol/L)/min (not statistically different from the SC administration). Conclusions: A simple, valid, sensitive, and inexpensive method for the measurement of gemcitabine in serum has been developed. This method may be useful for monitoring gemcitabine levels in cancer patients as part of therapeutic drug monitoring.

Characteristics of mildly refined soy protein fractions for cleaner soy‐food development

SummaryIn recent years, the markets for plant‐based products and plant protein have experienced a remarkable global expansion. The demand for plant‐based products, ranging from meat substitutes and dairy alternatives to plant‐based snacks and beverages, has surged exponentially. Soy protein, among all the plant protein resources, held a significant market share in the plant‐based industry, reflecting its popularity and widespread use. The selection of an appropriate soy fractionation process is essential for the development of cleaner and more sustainable soy‐based food products. This study compared three different soy protein fractions (SPFs) obtained through mild fractionation processes, each with unique chemical compositions and functionalities. Aqueous ethanol washing process resulted in the fractions with the highest protein purity and water holding capacity, while the simplified aqueous extraction method coupled with either an alkaline solubilisation step (SAE‐Alkaline) or an acid precipitation step (SAE‐Acid) yielded protein fractions with superior solubility, thermal stability, and gel strength. The choice of fractionation route has significant implications for the production of protein‐rich ingredients, and prioritising sustainable processing methods can contribute to the development of innovative and environmentally friendly soy‐based foods.

Determination of succinic acid and lactic acid in pigs' serum, intestinal contents, and meat by ultrahigh-performance liquid chromatography-tandem mass spectrometry.

Succinic acid and lactic acid have been associated with diarrhea in weaned piglets. The level of succinic acid and lactic acid in serum, meat, and intestinal contents is important to elucidate the mechanism of diarrhea in weaned piglets. A facile method was developed for the quantification of succinic acid and lactic acid in pigs' serum, intestinal contents, and meat using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC/MS/MS). The serum samples underwent protein precipitation with methanol. The meat and intestinal contents were freeze-dried and homogenized using a tissue grinding apparatus. Methanol-water mixture (80:20, v/v) was used for homogenizing the meat, while water was used for homogenizing the intestinal contents. An additional step of protein precipitation with acetonitrile was required for the intestinal contents. The resulting solution was diluted with water before being analyzed by UHPLC/MS/MS. Separation of succinic acid and lactic acid could be achieved within 3min using a Kinetic XB-C18 column. The coefficients of variation for peak areas of succinic acid and lactic acid were less than 5.0%. The established method demonstrated good linearity as indicated by correlation coefficients exceeding 0.996. Additionally, satisfactory recoveries ranging from 88.58% to 108.8% were obtained. The detection limits (RS/N = 3) for succinic acid and lactic acid were determined to be 0.75 ng/mL and 0.02 μg/mL, respectively. This method exhibited high sensitivity, simplicity in operation, and small sample weight, making it suitable for quantitative determination of succinic acid and lactic acid in pigs' serum, intestinal contents, and meat. The method developed will provide valuable technical support in studying the metabolic mechanisms of succinic acid and lactic acid in pigs.

Apixaban Pharmacokinetics and Bioequivalence of Two Tablet Formulations: A Randomized, Open-Label, Crossover Study, Fasting Condition in Healthy Indonesian Volunteers.

The present study aimed to assess the bioequivalence of a new apixaban generic with reference formulation. Twenty-six healthy volunteers were recruited for an open-label, balanced, randomized, 2-treatment, 2-sequence, 2-period, single oral dose study. Following overnight fasting, each volunteer received 5mg of apixaban test and reference formulations as single doses, separated by a 1-week washout period. Twenty blood samples were collected at predose and multiple time points between 0.5 and 72 hours after dosing. A validated ultra-performance liquid chromatography-tandem mass spectrometry detection method following a protein precipitation step was implemented to determine apixaban concentrations. Noncompartmental analysis was used to derive the pharmacokinetic parameters, which were then compared between the test and reference products using a multivariate analysis of variance. The pharmacokinetic parameters of the test product were not statistically different from the reference product, and the 90% confidence intervals of apixaban natural log-transformed area under the concentration-time curve from time 0 to infinity, area under the concentration-time curve from time 0 to the last measurable concentration, and maximum concentration were within 80%-125% based on the bioequivalence acceptance range criteria. The test and reference formulations of apixaban are bioequivalent in healthy subjects under fasting conditions.

Hydrometallurgical Method of Producing Lithium Perrhenate from Solutions Obtained during the Processing of Li-Ion Battery Scrap

The work presents the research results regarding the development of an innovative technology for the production of lithium perrhenate. The new technology is based entirely on hydrometallurgical processes. The source of lithium was solutions created during the processing of Li-ion battery masses, and the source of rhenium was perrhenic acid, produced from the scraps of Ni-based superalloys. The research showed that with the use of lithium carbonate, obtained from post-leaching solutions of Li-ion battery waste and properly purified (by washing with water, alcohol, and cyclic purification with CO2), and perrhenic acid, lithium perrhenate can be obtained. The following conditions: room temperature, time 1 h, 30% excess of lithium carbonate, and rhenium concentration in the acid from 20 g/dm3 to 300 g/dm3, allowed to produce a compound containing a total of 1000 ppm of metal impurities. The developed technology is characterized by the management of all aqueous waste solutions and solid waste and the lack of loss of valuable metals such as rhenium and lithium after the initial precipitation step of lithium carbonate.

Process Development of a Macrocyclic Peptide Inhibitor of PD-L1.

This article outlines the process development leading to the manufacture of 800 g of BMS-986189, a macrocyclic peptide active pharmaceutical ingredient. Multiple N-methylated unnatural amino acids posed challenges to manufacturing due to the lability of the peptide to cleavage during global side chain deprotection and precipitation steps. These issues were exacerbated upon scale-up, resulting in severe yield loss and necessitating careful impurity identification, understanding the root cause of impurity formation, and process optimization to deliver a scalable synthesis. A systematic study of macrocyclization with its dependence on concentration and pH is presented. In addition, a side chain protected peptide synthesis is discussed where the macrocyclic protected peptide is extremely labile to hydrolysis. A computational study explains the root cause of the increased lability of macrocyclic peptide over linear peptide to hydrolysis. A process solution involving the use of labile protecting groups is discussed. Overall, the article highlights the advancements achieved to enable scalable synthesis of an unusually labile macrocyclic peptide by solid-phase peptide synthesis. The sustainability metric indicates the final preparative chromatography drives a significant fraction of a high process mass intensity (PMI).