Internal Standard an Important Analyte Use in Drug Analysis by Liquid Chromatography Mass Spectrometry- An Article

Internal standard is an external compound which is mixed with targeted analytical solution and matrix as a constant concentration and use for preparing calibration standard curve by using ratio of analyte area and internal standard area with analyte concentration and internal standard concentration. This calibration curve used for quantification of unknown concentration of anlayte of interest. This article provide necessary information about internal standard like its selection procedure, characterization, types and response factor , to all analyst who are connected with drug analysis. This article is more important and I think first article which focuses a clear idea about internal standard use in drug analysis.

Studies of the pharmacokinetics of morroniside and loganin in rat after oral administration of raw and wine-processed Corni fructus by UPLC-QqQ-MS/MS

Purpose: To study the pharmacokinetics of morroniside (MR) and loganin (LG) in rats after oral administration of raw and wine-processed Corni fructus by UPLC-QqQ-MS/MS. Methods: Arctiin (AT) was used as internal standard, and the plasma or tissue samples were extracted twice using ethyl acetate. Electrospray ionization (ESI) negative ion mode was used, and the multiple reaction monitoring mode (MRM) was set in acquisition mode. The extraction and detection method is supported by selectivity, sensitivity, precision, accuracy, stability, extraction, recovery and matrix effect. The non-atrioventricular model of das2.0 software was used to calculate the pharmacokinetic parameters. Results: The absorption rate of MR (PTmax=0.092) and LG (PTmax=0.092) in Corni Fructus after wine-processing was faster in rats. The mean residence time was longer, and distribution of MR (PMRT0~t = 0.294) and LG (PMRT0~t = 0.000) in wine-processed Corni Fructus group increased in liver and kidneys. Conclusion: The proposed method has been successfully validated and is suitable for studying the pharmacokinetics of the two analytes in rats.

Mining the Wheat Grain Proteome

Bread wheat is the most widely cultivated crop worldwide, used in the production of food products and a feed source for animals. Selection tools that can be applied early in the breeding cycle are needed to accelerate genetic gain for increased wheat production while maintaining or improving grain quality if demand from human population growth is to be fulfilled. Proteomics screening assays of wheat flour can assist breeders to select the best performing breeding lines and discard the worst lines. In this study, we optimised a robust LC–MS shotgun quantitative proteomics method to screen thousands of wheat genotypes. Using 6 cultivars and 4 replicates, we tested 3 resuspension ratios (50, 25, and 17 µL/mg), 2 extraction buffers (with urea or guanidine-hydrochloride), 3 sets of proteases (chymotrypsin, Glu-C, and trypsin/Lys-C), and multiple LC settings. Protein identifications by LC–MS/MS were used to select the best parameters. A total 8738 wheat proteins were identified. The best method was validated on an independent set of 96 cultivars and peptides quantities were normalised using sample weights, an internal standard, and quality controls. Data mining tools found particularly useful to explore the flour proteome are presented (UniProt Retrieve/ID mapping tool, KEGG, AgriGO, REVIGO, and Pathway Tools).

Development and Greenness Assessment of HPLC Method for Studying the Pharmacokinetics of Co-Administered Metformin and Papaya Extract

Foods with medical value have been proven to be beneficial, and they are extensively employed since they integrate two essential elements: food and medication. Accordingly, diabetic patients can benefit from papaya because the fruit is low in sugar and high in antioxidants. An RP-HPLC method was designed for studying the pharmacokinetics of metformin (MET) when concurrently administered with papaya extract. A mobile phase of 0.5 mM of KH2PO4 solution and methanol (65:35, v/v), pH = 5 ± 0.2 using aqueous phosphoric acid and NaOH, and guaifenesin (GUF) were used as an internal standard. To perform non-compartmental pharmacokinetic analysis, the Pharmacokinetic program (PK Solver) was used. The method’s greenness was analyzed using two tools: the Analytical GREEnness calculator and the RGB additive color model. Taking papaya with MET improved the rate of absorption substantially (time for reaching maximum concentration (Tmax) significantly decreased by 75% while maximum plasma concentration (Cmax) increased by 7.33%). The extent of absorption reduced by 22.90%. Furthermore, the amount of medication distributed increased (30.83 L for MET concurrently used with papaya extract versus 24.25 L for MET used alone) and the clearance rate rose by roughly 13.50%. The results of the greenness assessment indicated that the method is environmentally friendly. Taking papaya with MET changed the pharmacokinetics of the drug dramatically. Hence, this combination will be particularly effective in maintaining quick blood glucose control.

Eco-Friendly, Simple, Fast, and Sensitive UPLC-MS/MS Method for Determination of Pexidartinib in Plasma and Its Application to Metabolic Stability

Pexidartinib is the first drug approved by the U.S. Food and Drug Administration specifically to treat the rare joint tumor tenosynovial giant cell tumor. In the current study, a validated, selective, and sensitive UPLC-MS/MS assay was developed for the quantitative determination of pexidartinib in plasma samples using gifitinib as an internal standard (IS). Pexidartinib and IS were extracted by liquid-liquid extraction using methyl tert-butyl ether and separated on an acquity BEH C18 column kept at 40 °C using a mobile phase of 0.1% formic acid in acetonitrile: 0.1% formic acid in de-ionized water (70:30). The flow rate was 0.25 mL/min. Multiple reaction monitoring (MRM) was operated in electrospray (ESI)-positive mode at the ion transition of 418.06 > 165.0 for the analyte and 447.09 > 128.0 for the IS. FDA guidance for bioanalytical method validation was followed in method validation. The linearity of the established UPLC-MS/MS assay ranged from 0.5 to 1000 ng/mL with r > 0.999 with a limit of quantitation of 0.5 ng/mL. Moreover, the metabolic stability of pexidartinib in liver microsomes was estimated.

Development and Validation of a Bioanalytical UHPLC-MS/MS Method Applied to Murine Liver Tissue for the Determination of Indocyanine Green Loaded in H-Ferritin Nanoparticles

Indocyanine green (ICG) is one of the most commonly used fluorophores in near-infrared fluorescence-guided techniques. However, the molecule is prone to form aggregates in saline solution with a limited photostability and a moderate fluorescence yield. ICG was thus formulated using protein-based nanoparticles of H-ferritin (HFn) in order to generate a new nanostructure, HFn-ICG. In this study, an ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) system was employed to develop and validate the quantitative analysis of ICG in liver tissue samples from HFn-ICG-treated mice. To precipitate HFn, cold acetone in acidic solution at pH 5.0 was used. The processed liver samples were injected into the UHPLC-MS/MS system for analysis using the positive electrospray ionization mode. Chromatographic separation was achieved on a Waters Acquity UPLC® HSS T3 Column (1.8 μm, 2.1 × 100 mm) with 0.1% formic acid and acetonitrile as the mobile phase with gradient elution. The selected reaction monitoring transitions of m/z 753 →m/z 330 and m/z 827 →m/z 330 were applied for ICG and IR-820 (the internal standard, IS), respectively. The method was selective and linear over a concentration range of 50–1,500 ng/ml. The method was validated for sensitivity, accuracy, precision, extraction recovery, matrix effect, and stability in liver tissue homogenates. ICG extraction recoveries ranged between 85 and 108%. The intra- and inter-day precisions were less than 6.28%. The method was applied to a bio-distribution study to compare the amount of ICG levels from mice treated with HFn-ICG and free ICG. The analyses of the homogenate samples from the two types of treatment showed that the concentration levels of ICG is approximately six-fold higher than those of free ICG (1,411 ± 7.62 ng/ml vs. 235 ± 26.0 ng/ml) at 2 h post injection.

Adjuvant measures and genetic evaluations to improve results of hair transplantation

Follicular unit extraction (FUE) has evolved dramatically as the most recent advancement in surgical hair restoration as it leaves a tiny scar and creates natural and pleasing results. This study aims to show the effectiveness of adjuvant measures and genetic evaluations in improving outcomes. Prospective analysis of 271 male patients with androgenic alopecia who underwent hair transplantation with FUE technique between August 2015 and February 2020 at our center was conducted. The mean age was 35.93 ±4.40 years. At one year postoperatively, patients were asked to fill up a questionnaire which included their satisfaction level, need for 2nd session, and complications. Informed written consent was obtained from all patients. Also, blood samples were provided from patients before the operation. RNA extraction and cDNA synthesis were performed using the RNX-Plus kit (Cinnagen, Iran) and Vivantis kit (Malaysia). Amplification of SRD5A2 and GAPDH genes (as internal standard) for measuring gene expression was performed by real-time PCR. Data were analyzed using the statistical package for social science SPSS V. 23. In the last 156 cases, the addition of 40 mg of Triamcinolone to the LA solution led to a dramatic reduction of the incidence of postoperative oedema, from 40% to 9%. Adding three sessions of PRP at 2nd, 4th and 6th months postoperatively resulted in an increased patient satisfaction rate with better hair density and thickness where the rate of highly satisfied patients increased from 64.5% to 83.7%. The addition of 40 mg Triamcinolone to the LA solution was highly effective in reducing postoperative oedema. Three sessions of PRP at 2nd, 4th and 6th months postoperatively were recommended. The results of SRD5A2 gene expression showed that the expression of this gene in satisfied (P = 0.049) and dissatisfied (P = 0.028) patients were significantly higher than highly satisfied patients, which means that the SRD5A2 gene expression had an essential role in the successfulness of hair transplantation. The increased expression of this gene could reduce the response to hair transplantation.

The Strategic Imperative of Quality Assurance

This chapter focuses on the fundamental question: What is the strategic importance of quality assurance (QA) for higher education institutions (HEIs)? Divided into four main segments, the chapter begins by defining QA within the context of a continuum, ranging from entirely voluntarily QA—as a part of the HEI internal standard procedures—to more ‘prescriptive' models, as mandated by ranking and accreditation bodies. Secondly, the chapter engages in identifying specific strategic functions of QA in light of two inherent objectives of HEIs: competitiveness and sustainable impact. Next, the chapter showcases the role of QA played in advancing the American University in Cairo (AUC) School of Business to the renowned triple-crown accreditation status (AACSB, EQUIS, AMBA), while highlighting both challenges encountered and implementable solutions. Last of all, the chapter discusses potential future changes to QA, given the ‘new normal' of the higher education landscape.