{"title":"Evaluation of Jacalin lectin sorbents for the extraction of the human chorionic gonadotropin glycoforms prior to analysis by nano liquid chromatography-high resolution mass spectrometry","authors":"Anastasia Goumenou , Christophe Chendo , Audrey Combès , Thierry Fournier , Valérie Pichon , Nathalie Delaunay","doi":"10.1016/j.jpba.2024.116525","DOIUrl":"10.1016/j.jpba.2024.116525","url":null,"abstract":"<div><div>Human chorionic gonadotropin (hCG) is a dimeric, highly glycosylated hormone with a total of 4 N- and 4 O-glycosylation sites in its two subunits, hCGα and hCGβ. Recently, we developed a novel nano liquid chromatography coupled to high resolution mass spectrometry (nanoLC-HRMS) method for the analysis and thus the detection of the intact glycoforms of hCG. Here, a sorbent functionalized with the Jacalin lectin was evaluated in solid-phase extraction (SPE) for its potential to fractionate the hCG glycoforms prior to their nanoLC-HRMS analysis at the intact level, which may facilitate the detection of low-abundance glycoforms and may lead to a more detailed characterization of the hormone glycosylation. A commercial sorbent based on Jacalin immobilized on Sepharose and having a lectin density of 4.5 mg per ml of gel was selected to carry out SPE and its capacity was estimated to be of some tens of μg of hCG per ml of lectin sorbent. Next, the SPE protocol was modified to improve the extraction recoveries. Especially, it was noticed that an extensive pre-conditioning procedure prior to the first use of a cartridge was necessary to remove the residual non-grafted lectins. Indeed, if non-grafted lectins are not eliminated, they may bind a part of hCG glycoforms preventing their retention by the sorbent, leading to low extraction recoveries (around 10 %). With the extensive pre-conditioning procedure, the average extraction recoveries for both hCGα and hCGβ glycoforms were about 50 %, with either recombinant or urinary hCG. Qualitatively, the fractionation of hCG glycoforms between the washing and elution fractions was achieved with the urinary hCG sample by determining the number of glycoforms detected in each fraction. It appears that 12 hCGα glycoforms have a low affinity (detected only in the washing fraction), 1 a low-medium affinity (detected in washing and elution 1 fractions), 16 a medium affinity (detected in washing, elution 1 and 2 fractions), and 12 a high affinity (detected only in elution 1 and 2 fractions). For the hCGβ glycoforms, similarly, 3 have a low affinity and 12 a low-medium affinity. Additionally, the 3 hCGβ glycoforms were detected better. A different behavior was observed with the recombinant hCG sample, which indicates glycosylation differences between the two hCG samples. This shows the potential of lectin-based affinity fractionation before nanoLC-HRMS analysis to better characterize the glycosylation state of hCG at the intact level.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142502441","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Screening and verification of target and molecular docking study of Pien-Tze-Huang in ameliorating alcoholic liver injury in rats","authors":"Shouer Lin , Pingping Wu , Youjia Wu , Liying Huang , Lingyi Huang","doi":"10.1016/j.jpba.2024.116517","DOIUrl":"10.1016/j.jpba.2024.116517","url":null,"abstract":"<div><div>Pien-Tze-Huang (PTH) is a famous traditional Chinese patent medicine with excellent liver-protection effects. However, the mechanism of hepatoprotective action has not yet been entirely elucidated. This study aimed to elucidate the protective mechanism of PTH against alcoholic liver injury in rats from key targets. An alcoholic liver disease (ALD) model in male rats was established, and the rats were treated with PTH given at a prescribed dosage. The hepatoprotective components of PTH and their exposure in the serum of PTH-treated rats were systematically identified. Quantitative proteomics was employed to find differentially expressed proteins. The key targets were screened by bioinformatic analysis and further validated by Western blotting (WB) and molecular docking. Ursodeoxycholic acid, notoginsenoside R1, gypenoside XVII, ginsenoside Rb1, and ginsenoside Re may be important active hepatoprotective components of PTH. A total of 53 differentially expressed proteins that were reversed by PTH were successfully identified in rat liver tissues. Retinol metabolism and the PPAR signaling pathway may play a key role in ameliorating alcohol-induced liver injury after PTH intervention. In particular, protein CYP2, FATCD36, FATP, ACS, and CPT-2 in these two pathways may be key targets for the therapeutic effects of PTH, with the same reversal observed by WB. Molecular docking analysis further revealed that these five proteins exhibited generally stable binding with the five main components of PTH. The hepatoprotective effects of PTH may be exerted through the modulation of key targets within pivotal pathways. This work pioneered a comprehensive screening of the active compounds in PTH and elucidated the mechanisms and targets of their protective effects against alcoholic liver injury, providing a reference for the broader clinical application of PTH.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142502478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xiaoyu Ma , Yongping He , Diya Lv , Xiaofei Chen , Zhanying Hong , Yifeng Chai , Yue Liu
{"title":"Optimization of metabolomics pretreatment method of cholangiocarcinoma cells based on ultrahigh performance liquid chromatography coupled with mass spectrometry","authors":"Xiaoyu Ma , Yongping He , Diya Lv , Xiaofei Chen , Zhanying Hong , Yifeng Chai , Yue Liu","doi":"10.1016/j.jpba.2024.116508","DOIUrl":"10.1016/j.jpba.2024.116508","url":null,"abstract":"<div><div>Metabolomics intends to maximize the quantity of available metabolites for the global metabolome, which largely depends on sample pretreatment protocols. However, there are few studies that comprehensively examined the effects of extraction and reconstitution solvents on metabolome coverage of adherent mammalian cells. In this study, the human cholangiocarcinoma TFK-1 cells were chosen as a cell model, and eight extraction solvents and five reconstitution solvents were used for the pretreatment based on ultrahigh performance liquid chromatography coupled with mass spectrometry (UPLC/MS). The coverage, reproducibility, and stability of the data were norms to evaluate the effectiveness of different extraction solvents and reconstitution solvents. Based on the number of metabolites, the mean Euclidean distance (ED<sub>MEAN</sub>) in the principal component analysis (PCA) 3D score plots and the relative standard deviation (RSD) distribution of metabolites, it was demonstrated that MeOH-CHCl<sub>3</sub>-H<sub>2</sub>O (8:1:1, v/v/v) was the optimal extraction solvent and MeOH-H<sub>2</sub>O (1:1, v/v) or H<sub>2</sub>O was superior to other reconstitution solvents for RP column analysis, and the extraction solvent MeOH-ACN-H<sub>2</sub>O (2:2:1, v/v/v) and the reconstitution solvents ACN-H<sub>2</sub>O (4:1, v/v) or MeOH-H<sub>2</sub>O (1:1, v/v) provide the best performance for HILIC column analysis. The optimized pretreatment methods explored in this study expand the coverage of polar and non-polar metabolites and improve the reproducibility and stability of the metabolic data, which can be applied to UPLC/MS-based global metabolomics study on cholangiocarcinoma cells, potentially providing better extraction solvents and reconstitution solvents for other adherent mammalian cells with similar chemical and physical properties.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142468219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yingfei Wang , Shijiao Zhang , Yingying Ge , Chunxia Miao , Benrong Liu , Tao Yang , Xiangjun Qiu , Wenchao Ou
{"title":"Serum untargeted metabolomics analysis of mice after myocardial infarction affected by qiliqiangxin capsule","authors":"Yingfei Wang , Shijiao Zhang , Yingying Ge , Chunxia Miao , Benrong Liu , Tao Yang , Xiangjun Qiu , Wenchao Ou","doi":"10.1016/j.jpba.2024.116516","DOIUrl":"10.1016/j.jpba.2024.116516","url":null,"abstract":"<div><div><em>Qiliqiangxin</em> (QLQX) capsule consists of 11 herbs, namely Huang qi (<em>astragalus membranaceus</em>), Ren shen (g<em>inseng</em>), Fu zi (<em>radix aconiti carmichaeli</em>)<em>,</em> Dan shen (<em>salvia miltiorrhiza</em>), Ting li zi (<em>lepidium seed</em>), Ze xie (<em>rhizoma alismatis</em>), Yu zhu (<em>radix</em> polygonati of<em>ficinalis</em>), Gui zhi (<em>cassia twig</em>), Hong hua (<em>carthamus tinctorious</em>), Xiang jia Pi (<em>cortex periploca</em>e), Chen Pi (<em>pericarpium citri reticulatae</em>), and it is a standardized commercial formula designed to address yang deficiency and to restore the balance of qi in the heart. QLQX is also known to invigorate the blood and promote the circulation of the blood and to promote the use of fluids to relieve water retention and edema, and can be used in cardiovascular diseases such as mild to moderate congestive heart failure resulting from coronary artery disease and hypertension. The further research on the effect of QLQX on cardiac function in mice after myocardial infarction was manipulated. QLQX was given to mice in myocardial infarction model by gavage with appropriate dosage and the samples were analyzed at the end of the animal experiments through the UHPLC-Q-Exactive LC-MS. The liquid mass spectrometry was used to collect and followed by further analysis of the corresponding metabolites and metabolic pathways using metabolomics analysis. As a result, 9 differential metabolites were identified, with 15 being up-regulated and 4 down-regulated following intervention with QLQX. Then the metabolic pathways by KEGG enrichment pathway bubble diagram was analyzed, and 4 metabolic pathways were obtained, and combined with the metabolites that had been screened and analyzed together, finally the two differential metabolites, 2,5-Dihydroxybenzenesulfonic Acid and o-Cresol sulfate were found. The Glycerophospholipid metabolism pathway was closely related to the remaining seven differential metabolites, and the pathway might be an important pathway related to the effects of QLQX on cardiac function in mice.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142432582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Piaopiao Qiu , Aihua Xia , Xinying Yang , Lin Yi , Yilan Ouyang , Yiming Yao , Haiying Liu , Liang Li , Zhenqing Zhang
{"title":"Metabolomic analysis reveals the potential of fucosylated chondroitin sulfate from sea cucumber in modulating metabolic homeostasis","authors":"Piaopiao Qiu , Aihua Xia , Xinying Yang , Lin Yi , Yilan Ouyang , Yiming Yao , Haiying Liu , Liang Li , Zhenqing Zhang","doi":"10.1016/j.jpba.2024.116509","DOIUrl":"10.1016/j.jpba.2024.116509","url":null,"abstract":"<div><div>In this study, we prepared four derivatives of fucosylated chondroitin sulfate (FCS): full-length FCS (flFCS) from <em>Holothuria leucospilota</em>, low molecular weight FCS (lmFCS) derived from flFCS, and their de-branched counterparts, de-branched flFCS (d-flFCS) and de-branched lmFCS (d-lmFCS) via controlled acid treatment. Following structural verification using various analytical techniques, we applied targeted metabolomics to examine the impact of FCS on nutritional efficacy and its structure-activity relationship. Analysis of 225 plasma and feces samples from 75 mice revealed a positive correlation between metabolomic shifts and increased weight gain, underscoring FCS’s potential to enhance nutrient absorption and promote growth. The observed linear relationship between the levels of short-chain fatty acids in plasma and feces suggests that FCS may facilitate catabolic activities in the gastrointestinal tract. The comparative study of different FCS derivatives on mouse growth and metabolic homeostasis regulation led to the conclusion that FCS exhibits greater biological activity with a higher degree of branching and larger molecular weight.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142446663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xiaodong Wu , Ling Qin , Miao Song , Chuanming Zhang , Jingjing Guo , Zheng Yang , Zhixiang Gao , Min Qiu
{"title":"Metagenomics combined with untargeted metabolomics to study the mechanism of miRNA-150-5p on SiO2 -induced acute lung injury","authors":"Xiaodong Wu , Ling Qin , Miao Song , Chuanming Zhang , Jingjing Guo , Zheng Yang , Zhixiang Gao , Min Qiu","doi":"10.1016/j.jpba.2024.116515","DOIUrl":"10.1016/j.jpba.2024.116515","url":null,"abstract":"<div><div>Acute lung injury is a significant global health issue, and its treatment is becoming a hot topic of the researchers. To investigate the feasibility of miRNA-150–5p tail vein injection in the treatment of SiO<sub>2</sub>-induced acute lung injury through the regulation of gut microbiota and serum metabolites based on multiomics technology. Twenty-four mice were randomly divided into the control, SiO<sub>2</sub> and miRNA-150–5p intervention groups. The SiO<sub>2</sub> and miRNA-150–5p intervention groups received a single intranasal dose of 100 µL 4 % SiO<sub>2</sub> suspension. Meanwhile, the miRNA-150–5p intervention group was administered with two tail vein injections of miRNA-150–5p (15 nmol each per mouse) on the day of successful modelling and on the third day post modelling. Metagenomics and metabolomics techniques were used to measure gut microbiota and serum metabolites, respectively. Tail vein injection of miRNA-150–5p improved SiO<sub>2</sub>-induced acute lung injury and reduced the secretion of inflammatory factors interleukin (IL)-6, tumour necrosis factor-α and IL-1β. These conditions altered the structure of gut microbiota, which resulted in the notable modulation of eight species at the species level. In addition, tail vein injection of miRNA-150–5p considerably reduced the levels of substances, such as phosphatidylethanolamine, phosphatidylcholine and phosphatidylinositol, in the glycerophospholipid metabolism and glycosylphosphatidylinositol–anchor biosynthesis pathways. Tail vein injection of miRNA-150–5p can alleviate acute lung injury. Combined metagenomics and untargeted metabolomics revealed the miRNA-150–5p-mitigated SiO<sub>2</sub>-induced acute lung injury that occurred through the regulation of gut microbiota and serum metabolites.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142441583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
David M. Saylor, Robert M. Elder, Kaleb Duelge, Nimesh P.R. Ranasinghe Arachchige, David D. Simon, Samanthi Wickramasekara, Joshua A. Young
{"title":"Inter-laboratory study for extraction testing of medical devices","authors":"David M. Saylor, Robert M. Elder, Kaleb Duelge, Nimesh P.R. Ranasinghe Arachchige, David D. Simon, Samanthi Wickramasekara, Joshua A. Young","doi":"10.1016/j.jpba.2024.116496","DOIUrl":"10.1016/j.jpba.2024.116496","url":null,"abstract":"<div><div>Biocompatibility evaluation of medical devices often relies on chemical testing according to ISO 10993–18 as a critical component for consideration. However, the precision associated with these non–targeted chemical characterization assessments has not been well established. Therefore, we have conducted a study to characterize intra–laboratory (repeatability) and inter–laboratory (reproducibility) variability associated with chemical testing of extractables from polymeric materials. To accomplish this, this study focused on two polymers, each with nine chemicals that were intentionally compounded into the materials. Eight different laboratories performed extraction testing in two solvents and subsequently characterized the extracts using gas chromatography and liquid chromatography methods. Analysis of the resulting data revealed the central 90 % range for the repeatability and reproducibility relative standard deviations are (0.09, 0.22) and (0.30, 0.85), respectively, for the participating laboratory methods. This finding implies that if the same sample was tested by two different laboratories using the same extraction conditions, there is 95 % confidence for 95 % of systems that the test results could exhibit differences up to 240 %. While the study was not designed to evaluate the relative impact of specific underlying factors that may contribute to variability in quantitation, the data obtained suggest the variability associated with analytical method alone is a substantial contribution to the overall variability. The relatively large reproducibility limits we observed may have significant implications where variability in extraction measurements can impact aspects of biocompatibility risk evaluation, such as exposure dose estimation and chemical equivalence assessments.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142432583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hesham M. El-Sayed , Dalia M. Rasheed , Engy A. Mahrous , Basma M. Eltanany , Zeinab M. Goda , Laura Pont , Fernando Benavente , Essam Abdel-Sattar
{"title":"Metabolomics analysis of Cucumis melo var. flexuosus organs in correlation to its anti-inflammatory activity aided by chemometrics","authors":"Hesham M. El-Sayed , Dalia M. Rasheed , Engy A. Mahrous , Basma M. Eltanany , Zeinab M. Goda , Laura Pont , Fernando Benavente , Essam Abdel-Sattar","doi":"10.1016/j.jpba.2024.116512","DOIUrl":"10.1016/j.jpba.2024.116512","url":null,"abstract":"<div><div>Snake melon (<em>Cucumis melo</em> var. <em>flexuosus</em>, CM) is a gourd with health-promoting nutritional traits and unexplored phytochemicals. This study aims to comprehensively investigate the phytoconstituents in the fruits, leaves, roots, seeds, and stems of CM, using liquid chromatography-quadrupole time-of-flight tandem mass spectrometry. Consequently, 118 metabolites were identified, encompassing phenolic compounds, flavonoids, megastigmanes, lignans, cucurbitacins, and fatty acids. Multivariate data analysis revealed differences in the metabolite composition of CM organs and correlated these variations with the potential <em>in-vitro</em> anti-inflammatory properties assessed against RAW 264.7 macrophages through the down-regulation of cyclo-oxygenase-Ⅱ, nuclear factor-kappa B, and tumor necrosis factor-<em>α</em>. The results indicated that leaf and seed extracts showed the highest anti-inflammatory activity due to their enrichment in several flavonoids, phenolic glycosides, and a megastigmane. These findings emphasize the health benefits of CM organs as potential functional foods and functional food by-products, serving as a natural source for developing new anti-inflammatory agents.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142423402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hyeon-Jeong Lee , Jiin Hwang , Yoondam Seo , Gahyeon Lee , Hwa Jeong Lee , Hophil Min
{"title":"Simultaneous detection of myostatin-targeting monoclonal antibodies in dried blood spots and plasma using liquid chromatography-tandem mass spectrometry with field asymmetric ion mobility spectrometry","authors":"Hyeon-Jeong Lee , Jiin Hwang , Yoondam Seo , Gahyeon Lee , Hwa Jeong Lee , Hophil Min","doi":"10.1016/j.jpba.2024.116518","DOIUrl":"10.1016/j.jpba.2024.116518","url":null,"abstract":"<div><div>Transforming growth factor-β superfamily members, such as myostatin, growth/differentiation factor 11, and activin A, negatively regulate skeletal muscle mass. Inhibitors targeting these cytokines or activin receptor type IIB have the potential to treat muscular diseases and enhance physical performance. However, because of their effects on muscle mass and potential misuse, they are strictly prohibited in sports. Given the high potential for misuse as a doping agent in sports, effective analytical methods for these prohibited antibodies targeting these specific cytokines or their receptor are critically needed. In this study, we aimed to develop and validate a multitarget method to detect the prohibited transforming growth factor-β superfamily-targeting monoclonal antibodies, such as landogrozumab, domagrozumab, and the activin receptor type IIB-targeting antibody, bimagrumab, in human plasma and dried blood spot (DBS) samples using liquid chromatography-tandem mass spectrometry. Antibodies were purified from both the DBS and plasma samples using protein G magnetic beads and field-asymmetric ion mobility spectrometry (FAIMS) to minimize interference, followed by liquid chromatography-tandem mass spectrometry analysis. The validation process included tests for specificity, selectivity, linearity, limit of detection (LOD), limit of identification, precision, recovery, carryover effect, and matrix effect. The LODs for the target antibodies were identical in both DBS and plasma samples at 0.1 µg/mL for landogrozumab heavy and light chains, as well as 0.25 µg/mL for the domagrozumab light chain and 0.25 µg/mL for the bimagrumab heavy chain. However, the heavy chain of domagrozumab exhibited an LOD of 0.5 µg/mL in DBS and 1 µg/mL in plasma. The analytical method demonstrated strong linearity, with R² values greater than 0.99 for both plasma and DBS, and no carryover effect. Precision (CV%) was below 15 % at both middle (1 or 5 µg/mL; specific to the heavy chain of domagrozumab in plasma) and high (10 µg/mL) concentrations and was less than 20 % at the LOD. The selectivity and specificity indicated no interference in the analysis of target mAbs in different blood samples. Recovery was 31.6–49.8 % for DBS and 51.4–85.3 % for plasma, with no significant matrix effect. This study provides an effective method for doping analysis and novel protein detection.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142423389","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhiru Zhang , Rong Yue , Yibo Wang , Lizhou Ma , Miao Wang , Yu Chen
{"title":"To explore the mechanism of gypenosides in the treatment of liver injury in rats based on GC-MS metabolomics and bile acid metabolism pathway","authors":"Zhiru Zhang , Rong Yue , Yibo Wang , Lizhou Ma , Miao Wang , Yu Chen","doi":"10.1016/j.jpba.2024.116506","DOIUrl":"10.1016/j.jpba.2024.116506","url":null,"abstract":"<div><div>Gynostemma pentaphyllum is a herbaceous vine of Cucurbitaceae family, and its principal pharmacological components, gypenosides (GPs), have been proved to be effective in various liver diseases. However, the mechanisms of GPs on liver injury are still to be studied for further. This investigation utilized the CCl<sub>4</sub>-induced liver injury rat model (LI) to comprehensively explore the mechanism of action of GPs in the treatment of chemical liver injury by comparing the metabolomic changes in four groups rats. In this study, the therapeutic efficacy of GPs in a liver injury rat model induced by weekly gavage of CCl<sub>4</sub> was evaluated by inflammatory factors, oxidative damage indexes, and histopathological sections. Then, GC-MS technology was used to identify the metabolic profile of GPs in treating liver injury. Finally, the content variation of metabolites (BAs and SCFAs) was measured to elucidate the mechanism of GPs in the treatment of CCl<sub>4</sub>-induced liver injury. After 8 weeks of administration, GPs effectively reduced the degree of LI and appeared a substantial tendency of reversing in the levels of MDA, GSH, CYP7E1, CYP7A1 and CYP27A1. Untargeted metabolomics suggested that GPs may play a role in BAs and SCFAs metabolism. Targeted metabolomics and ELISA confirmed the key role of GPs in increasing SCFAs levels and regulating BAs metabolism. Overall, this study indicated that GPs can alleviate CCl<sub>4</sub>-induced liver injury. And GPs may exert beneficial effects on LI by affecting their metabolites (SCFAs and BAs).</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142441585","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}