Current Pharmaceutical Analysis - Volume 19, Issue 9, 2023

Quality by Design (QbD) is a systematic approach for improvement that stresses product and process and begins with a predetermined objective, as recommended by the USFDA and International Council Harmonization (ICH). Regulatory bodies frequently highlight the use of ICH quality criteria, which include Q8, Q9, Q10, and Q11. The differentiation between the traditional and QbD helps to study the risk assessment and technique for developing new products. There are a few steps involved in pharmaceutical and Analytical QbD. Various factors were used for the study of QbD, such as Analytical Target Product Profile (ATPP), Risk Assessment Quality Design Space, Control Strategy, etc. Critical Quality Attribute (CQA) may be understood and analyzed via a way of means of understanding the goods and technique and risk evaluation is useful for effective verbal exchange among FDA and industry, research/improvement and production, and amongst a couple of production sites inside the company. Life-cycle management of analytical procedure begins off evolving with the establishment of ATP and maintains until the approach is in use. The design of the experiment (DoE) involves the Q8 guidelines. DoE has been used in the rational development and optimization of analytical methods. Culture media composition, mobile phase composition, flow rate, and time of incubation are input factors (independent variables) that may be screened and optimized using DoE. Process analytical technology is implemented for the understanding and identification of developing a product and techniques. There are various benefits and applications of QbD in the pharmaceutical industry.

Aims: The aim of this project was to develop a QC friendly and efficient method of protein species of origin identification to replace more costly mass spectrometric based methods currently being used for this testing.Background: NMR relaxation measurements with proteins in aqueous solutions exploit the fast chemical exchange between water and exposed NH and OH protons of amino acid side chains in the folded protein structure unique to each biologic drug. Implementation of this technique has led to routine testing for authentication and forensics of biopharmaceuticals, determination of moisture content in lyophilized protein formulations and aggregation of proteins in solution. For small molecule applications, TD-NMR can detect if solvents are received neat or tainted with moisture, impurities, or denaturants.Objective: The objective of this study was to evaluate the ability of NMR Relaxation measurements to differentiate between sources of Albumin proteins as a rapid QC test. Evaluation of differences in molecular mobility between components in a solution as reflected in the longitudinal (T1) and transverse (T2) relaxation times of protons demonstrate that NMR relaxation techniques can distinguish between different albumin sources of origin.Methods: Representative albumin proteins from differing sources of origin were studied. Using bovine serum albumin (BSA) as the target species of origin, NMR relaxation techniques as well as chemometric modeling were used to evaluate the use of this technique for protein source of origin identification.Results: NMR Relaxation using benchtop instrumentation showed that the bovine albumin species of origin can be distinguished from porcine, chicken egg white and sheep sources of origin. Goat albumin selectivity remained questionable and BSA cannot be distinguished from human or rabbit sources of origin within the representative variability.T2 transverse relaxation results were significantly more discriminating for protein source identification than the T1 longitudinal relaxation result by itself. The T1 longitudinal relaxation result did not contribute significantly to this investigation. However, fusing the T1 data with the T2 transverse relaxation results and using larger data sets merit further evaluation in the hope of achieving additional selectivity.Conclusion: While additional lots are needed for more definitive results, this preliminary evaluation of using NMR Relaxation demonstrated the capability for the source of origin species discrimination and identification using benchtop NMR instrumentation.

Objective: This study aimed to investigate the main metabolites and metabolic pathways of oleraisoquinoline in rats, a new alkaloid isolated from Portulaca oleracea L., and test its antioxidation and anticholinesterase effects.Methods: Ultra-high-performance liquid chromatography-electrospray coupled with quadrupole time-of-flight mass spectrometry (UHPLC-ESI-Q-TOF/MS) was applied to study the metabolism of oleraisoquinoline. Furthermore, 1,1-diphenyl-2-picrylhydrazyl assay and modified Ellman's method were used to test the antioxidation and anticholinesterase effects of oleraisoquinoline, respectively.Results: The metabolism results of oleraisoquinoline showed, after its administration through the tail vein of rats, 4 metabolites in the plasma samples, 17 metabolites in the urine sample, and 2 metabolites in the feces sample. The main metabolic pathways were hydrolyzation, oxidation, hydroxylation, sulfonation, glucuronidation, acetylation, and methylation. Additionally, IC50 values of antioxidant and anticholinesterase activities were 13.819 ± 0.005 μM and 10.551 ± 0.069 μM, respectively.Conclusion: 21 metabolites were found in the rat's plasma, urine, and feces samples, and the metabolic pathways included hydrolyzation, oxidation, hydroxylation, sulfonation, glucuronidation, acetylation, and methylation; among them, sulfonation was the main metabolic reaction. Meanwhile, oleraisoquinoline also showed extremely good antioxidant and anticholinesterase activities.

Background: Bellidifolin (BEL) has a decent enemy of myocardial fibrosis impact, and its preparation into nano-micelles can build security and great biocompatibility in vitro and in vivo. The pharmacokinetic assessment of BEL can be utilized as the reason for the security and viability of BEL in clinical use.Objective: This research aimed to establish an effective UPLC-MS/MS strategy for assuring BEL in rodent plasma and concentrating on its pharmacokinetics in vivo.Methods: Luteolin was utilized as an internal standard (IS). Chromatographic separation was accomplished utilizing a UPLC HSS T3 column (2.1 æ#151;100 mm, 1.8 μm) section using a mobile phase of 0.1% acetonitrile (A) and 0.1% formic acid in water (B) with gradient elution. Electrospray ionization (ESI) coupled mass spectrometry was applied in various response checking (MRM) modes with negative ionization.Results: The pharmacokinetic behaviour of bellidifolin nano-micelles in vivo showed that the peak concentration (Cmax) was 1666.19±479.92 μg/L, the time to peak (Tmax) was 0.167 h, and the apparent elimination half-life (t1/2) was 7.60±3.58 h. The plasma clearance rate (CL/F) was 1.15±0.48 L/h/kg, the apparent volume of distribution (V/F) was 14.38±11.04, the area under the curve (AUC) was 8292.57±4193.13 μg/L*h, and the mean retention time (MRT) was 9.70±4.55 h.Conclusion: The method was successfully applied to the plasma pharmacokinetics of bellidifolin nano-micelles after intragastric administration to rats.

Introduction: Th study aims to establish the chromatographic fingerprint of Zhachong Shisan pills (ZSP), and to explore its active components and mechanism of analgesic and anti-inflammatory action.Method: First, the Personal Compound Database and Library (PCDL) of ZSP was constructed through literature mining, and then the chemical composition and fingerprint of methanol extractions from 8 batches of ZSP were studied using High-Performance Liquid Chromatography with Quadrupole Time-Of-Flight tandem Mass Spectrometry (HPLC-QTOF-MS) technology. Furthermore, network pharmacology was used to explore the active compounds, potential targets, and signal pathways of analgesic and anti-inflammatory activity of ZSP. A total of 102 compounds were detected in positive and negative ion mode.Results: Fifty-six characteristic peaks in positive ion mode and 78 characteristic peaks in negative ion mode were confirmed as common peaks in the fingerprint of ZSP. Through network pharmacology research, the effective components, key targets, and signal pathways of ZSP for the treatment of cerebral apoplexy by analgesic and anti-inflammatory effects were all analyzed.Conclusion: This study explained the substance basis of the analgesic and anti-inflammatory effects of Zhachong Shisan Pills, and explores its possible mechanisms, providing ideas for rational clinical medication and in-depth pharmacological research.