Journal of pharmaceutical and biomedical analysis最新文献_第2页

Detection of 11-nor-9-carboxy-hexahydrocannabinol (HHC-COOH) as metabolite of both hexahydrocannabinol (HHC) and Δ9-tetrahydrocannabinol (Δ9-THC) in routine forensic samples

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-22 DOI: 10.1016/j.jpba.2025.116833

Marcel Grapp , Christoph Kaufmann , Andreas Peschel , Meike Potzscher , Marek Dziadosz , Lisa Marquenie , Jörg Teske

{"title":"Detection of 11-nor-9-carboxy-hexahydrocannabinol (HHC-COOH) as metabolite of both hexahydrocannabinol (HHC) and Δ9-tetrahydrocannabinol (Δ9-THC) in routine forensic samples","authors":"Marcel Grapp , Christoph Kaufmann , Andreas Peschel , Meike Potzscher , Marek Dziadosz , Lisa Marquenie , Jörg Teske","doi":"10.1016/j.jpba.2025.116833","DOIUrl":"10.1016/j.jpba.2025.116833","url":null,"abstract":"<div><div>Hexahydrocannabinol (HHC) is a phytocannabinoid that has been known since 1940, but has only recently appeared on the recreational drug market. For the analytical detection of HHC consumption, a GC-MS method for the quantification of cannabinoids was extended and validated by adding (<em>9S</em>)-HHC, (<em>9 R</em>)-HHC, (<em>9S</em>)-carboxy-HHC (HHC-COOH) and (<em>9 R</em>)-HHC-COOH. Both HHC and HHC-COOH epimers were chromatographically separated and the validation data were convincing for forensic toxicological routine analysis. This method was used to analyze 599 serum samples from forensic cases where cannabis use was suspected. Results were classified into three consumption groups: Δ<sup>9</sup>-THC only (n = 574), Δ<sup>9</sup>-THC and HHC (n = 19), and HHC only (n = 6). The concentration in serum was between 0.15 ng/mL to 14.4 ng/mL for (9 <em>R</em>)-HHC and 0.14 ng/mL to 5.76 ng/mL for (9<em>S</em>)-HHC. In all HHC positive samples, (9 <em>R</em>)-HHC-COOH could be detected in concentrations between 1.0 and 314 ng/mL. Of note, in the cases that tested positive for Δ<sup>9</sup>-THC only, (9 <em>R</em>)-HHC-COOH was also unambiguously detected in serum samples as a metabolite not only of HHC but also of Δ<sup>9</sup>-THC. In six urine samples of THC users that were examined by GC-MS and LC-MS/MS both epimers of HHC-COOH and their glucuronides could be detected. (<em>9S</em>)-HHC-COOH was the predominant epimer in urine which was not detected in serum. Results suggest that detection of HHC-COOH epimers alone cannot prove prior HHC consumption. With the data presented, we tentatively recommend a cut-off of 30 % of the (<em>9 R</em>)-HHC-COOH/THC-COOH ratio in serum to distinguish the intake of both substances from the intake of Δ<sup>9</sup>-THC alone.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"261 ","pages":"Article 116833"},"PeriodicalIF":3.1,"publicationDate":"2025-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143696922","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}

引用次数: 0

From bioinformatics to clinical application: A new strategy in CRP detection with peptide aptamer

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-21 DOI: 10.1016/j.jpba.2025.116820

Xiaona Zhao , Tong gong Liu , Hongfang Chen , Xi Chen , Liwen Zhu , Jie Wen , Dayong Gu

{"title":"From bioinformatics to clinical application: A new strategy in CRP detection with peptide aptamer","authors":"Xiaona Zhao , Tong gong Liu , Hongfang Chen , Xi Chen , Liwen Zhu , Jie Wen , Dayong Gu","doi":"10.1016/j.jpba.2025.116820","DOIUrl":"10.1016/j.jpba.2025.116820","url":null,"abstract":"<div><div>C-Reactive protein (CRP) is a key biomarker for evaluating inflammation levels and estimating cardiovascular risk. However, current CRP detection methods rely on monoclonal antibodies (mAb), which possess shortcomings such as a lengthy preparation cycle, high cost, and poor repeatability. To address these challenges, we explored the potential of peptide aptamers as an alternative to mAb for CRP detection. Using some bioinformatics approaches, we designed and optimized peptide aptamers, selecting the dominant peptide aptamer C9m (KWRWRFRLSR) through experimental validation for its specific recognition of CRP. We then established a sandwich ELISA detection system combining C9m with CRP mAb. This system demonstrated a detection limit of 22.275 ng/mL CRP and exhibited excellent specificity, with no cross-reactivity observed with human serum albumin or γ-globulin. The method also showed high reproducibility, with intra- and inter-assay coefficients of variation (CV) less than 15 %, meeting laboratory testing standards. Furthermore, comparison with clinically used immunoturbidimetry revealed high consistency (r = 0.9891).</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"261 ","pages":"Article 116820"},"PeriodicalIF":3.1,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143697999","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}

引用次数: 0

Characterization of pneumococcal conjugates in vaccine process development by multi-detection hydrodynamic chromatography

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-20 DOI: 10.1016/j.jpba.2025.116826

Xiujuan Jia , James Z. Deng , Michael A. Winters , Mellie June Paulines , Weidong Tong , Erin Cannon , Mirlinda Biba , Ping Zhuang

{"title":"Characterization of pneumococcal conjugates in vaccine process development by multi-detection hydrodynamic chromatography","authors":"Xiujuan Jia , James Z. Deng , Michael A. Winters , Mellie June Paulines , Weidong Tong , Erin Cannon , Mirlinda Biba , Ping Zhuang","doi":"10.1016/j.jpba.2025.116826","DOIUrl":"10.1016/j.jpba.2025.116826","url":null,"abstract":"<div><div>Pneumococcal conjugate vaccines (PCVs) are developed by conjugating pneumococcal polysaccharides to a carrier protein, such as CRM<sub>197</sub>. Size and molecular weight are important attributes of each monovalent conjugate in a PCV, making accurate monitoring of molecular weights crucial during the conjugation process. While size-exclusion chromatography (SEC) coupled with multi-angle light scattering (MALS), refractive index (RI), and ultraviolet (UV) detectors (SEC-MALS) is the gold standard used for absolute molecular weight characterization, this study presents the development of a multi-detection (MALS, UV and RI) hydrodynamic chromatography (HDC-MALS) method and its utility for comprehensive PCV characterization. The optimized HDC-MALS method is employed for in-depth understanding of vaccine conjugation process and effective characterization of heterogeneous, large conjugates through granular molar mass distribution analysis. Compared to other mild separation techniques such as field flow fractionation (FFF), HDC allows for high mobile phase flow rates without compromising separation efficiency, enabling faster run times that meet the demands of in-process control with rapid turn-around times.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"261 ","pages":"Article 116826"},"PeriodicalIF":3.1,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143682401","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}

引用次数: 0

Development and characterisation of a method for determining N-terminal heterogeneity in the human recombinant follitropin β-subunit

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-19 DOI: 10.1016/j.jpba.2025.116828

Xinyue Hu , Xuezhi Han , Yue Sun , Xiaoming Zhang , Yi Li , Lvyin Wang , Ping Lv , Chenggang Liang , Jing Li

{"title":"Development and characterisation of a method for determining N-terminal heterogeneity in the human recombinant follitropin β-subunit","authors":"Xinyue Hu , Xuezhi Han , Yue Sun , Xiaoming Zhang , Yi Li , Lvyin Wang , Ping Lv , Chenggang Liang , Jing Li","doi":"10.1016/j.jpba.2025.116828","DOIUrl":"10.1016/j.jpba.2025.116828","url":null,"abstract":"<div><div>Recombinant human follicle-stimulating hormone (rhFSH) consists of α- and β subunits linked by non-covalent bonds. Both subunits exhibit N-terminal heterogeneity, with the β-subunit exhibiting heterogeneity to a greater extent. In this study, we developed a comprehensive method for assessing β-subunit N-terminal heterogeneity based on a previously developed peptide mapping approach. The new method, which includes optimised sample preparation and reversed-phase high-performance liquid chromatography, facilitated the separation of integrated (NSCELTNITIAIEK) βL1 and truncated (CELTNITIAIEK) βL1 forms. The identity of the separated integrated and truncated forms was confirmed via tandem mass spectrometry. Optimised sample preparation streamlined the process by simultaneously performing trypsin digestion (with calcium chloride enhancement) and PNGase F deglycosylation for a total preparation time of just 1 day. The enzymatic digestion and sample preparation processes underwent several evaluations, including scaled-up sample preparation, sample preparation variability assessment and extended incubation stability studies. The sample preparation process demonstrated excellent robustness in all evaluations. Results consistently showed approximately 50 % N-terminal truncation of two amino acids in the β-subunit across different manufacturers’ samples, highlighting the utility of the method in assessing the batch-to-batch consistency and comparability of biosimilar products. This integrated approach combines peptide mapping and N-terminal heterogeneity analysis into a single, efficient workflow, thereby significantly enhancing quality control processes and enhancing the safety and efficacy of recombinant FSH products.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"261 ","pages":"Article 116828"},"PeriodicalIF":3.1,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143696921","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}

引用次数: 0

Application of mass spectrometry for the advancement of PROTACs

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-19 DOI: 10.1016/j.jpba.2025.116829

Yuechen Hao, Baoshuang Zhang, Ruibing Chen

引用次数: 0

Chiral analysis of ketamine enantiomers in human urine and hair: Application to authentic cases of ketamine use

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-18 DOI: 10.1016/j.jpba.2025.116824

Zhen Zhang , Yan Shi , Meiting Lin , Ping Xiang , Liying Zhou , Hejian Wu , Xin Wang

{"title":"Chiral analysis of ketamine enantiomers in human urine and hair: Application to authentic cases of ketamine use","authors":"Zhen Zhang , Yan Shi , Meiting Lin , Ping Xiang , Liying Zhou , Hejian Wu , Xin Wang","doi":"10.1016/j.jpba.2025.116824","DOIUrl":"10.1016/j.jpba.2025.116824","url":null,"abstract":"<div><div>Ketamine, which possesses important anesthesia and antidepressant properties, has been used clinically in its racemate form. In the 1990s, the single enantiomer S-ketamine began to be used for clinical use. In 2019, an antidepressant S-ketamine hydrochloride nasal spray hit the market. Therefore, a method for chiral analysis of ketamine is particularly important, as this could reveal the types of drugs taken by individuals. An ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for the separation of chiral ketamine in human urine and hair in this study. Urine (50 μL) was diluted in 950 μL of methanol. Approximately 20 mg of hair was extracted in methanol by cryogenic grinding. The method was applied to authentic urine, blood, and 45 hair samples. Following the deceased's use of esketamine prior to death, only S-ketamine was found in the blood and urine, suggesting that ketamine does not undergo enantiomeric inversion in <em>vivo</em>. The ratio of R-ketamine to S-ketamine in 45 hair samples from ketamine abusers ranged from 0.809 to 1.43, indicating that the ketamine available on the illegal drug market was predominantly in its racemate form. Furthermore, a significant difference (P = 0.039) was observed in the enantiomeric ratio of ketamine between hair samples from ketamine abusers in China and Myanmar. The enantiomeric ratios found in ketamine sourced from various origins are potentially attributable to region variations and distinct synthetic routes. These findings provide a basis for analyzing ketamine enantiomers.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"261 ","pages":"Article 116824"},"PeriodicalIF":3.1,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143682400","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}

引用次数: 0

Simultaneous determination of phenylalanine and tyrosine levels in human blood obtained by the dried spot technique for monitoring of patients with phenylketonuria

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-18 DOI: 10.1016/j.jpba.2025.116831

Michal Kopčil , Roman Kanďár

{"title":"Simultaneous determination of phenylalanine and tyrosine levels in human blood obtained by the dried spot technique for monitoring of patients with phenylketonuria","authors":"Michal Kopčil , Roman Kanďár","doi":"10.1016/j.jpba.2025.116831","DOIUrl":"10.1016/j.jpba.2025.116831","url":null,"abstract":"<div><div>Monitoring patients with phenylketonuria (PKU) requires accurately measuring phenylalanine and tyrosine levels in a small volume of blood samples obtained by the dried blood spot (DBS) technique. The aim was to study selected parameters influencing the quantitative results. Phenylalanine and tyrosine were extracted from DBS samples with methanol, and 5 internal standard introduction techniques were tested. Phenylalanine and tyrosine levels were measured in 6-mm discs punched from DBS, pre-punched 9-mm discs containing the entire DBS sample, and liquid blood by HPLC-MS-MS. Levels in 6-mm discs punched from DBS measured by HPLC-MS/MS were compared with those measured by the HPLC-FLD. The analytical parameters of the method are satisfactory, linearity in the range of 25–1200 μmol/L (LOD, LOQ and LLOQ values 0.2 μmol/L, 0.5 μmol/L and 3.8 μmol/L for phenylalanine, 0.5 μmol/L, 1.5 μmol/L and 5.1 μmol/L for tyrosine), within-run precision 1.8 %–3.7 % for phenylalanine, 1.9 %–2.7 % for tyrosine, between-run precision 4.7 %–5.9 % for phenylalanine, 4.1 %–5.4 % for tyrosine, recovery 93.8 %–100.4 % for phenylalanine and 93.7 %–99.1 % for tyrosine. Good agreement was found between phenylalanine and tyrosine concentrations in 6-mm discs punched from DBS (R = 0.896, p < 0.001, and R = 0.907, p < 0.001, respectively), pre-punched 9-mm discs containing the entire DBS sample (R = 0.960, p < 0.001, and R = 0.950, p < 0.001, respectively) and liquid blood, as well as between phenylalanine and tyrosine concentrations obtained by HPLC-MS/MS and HPLC-FLD (R = 0.968, p < 0.001, and R = 0.984, p < 0.001, respectively). The presented method is suitable for monitoring patients with PKU.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"260 ","pages":"Article 116831"},"PeriodicalIF":3.1,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687076","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}

引用次数: 0

Simultaneous determination of thirteen related substances in lenalidomide API by a stability-indicating RP-HPLC method

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-17 DOI: 10.1016/j.jpba.2025.116823

Zhiling Cao, Maolong Huang, Luye Bi, Ling Zhang, Siyi Xia, Yuhan Sun, Yang Yu, Xudong Yu, Dahua Sh, Fan Xu

{"title":"Simultaneous determination of thirteen related substances in lenalidomide API by a stability-indicating RP-HPLC method","authors":"Zhiling Cao, Maolong Huang, Luye Bi, Ling Zhang, Siyi Xia, Yuhan Sun, Yang Yu, Xudong Yu, Dahua Sh, Fan Xu","doi":"10.1016/j.jpba.2025.116823","DOIUrl":"10.1016/j.jpba.2025.116823","url":null,"abstract":"<div><div>The purpose of this study was to develop and validate a high performance liquid chromatography (HPLC) method for the separation and determination of related impurities and potential genotoxic impurity in the lenalidomide API. Chromatographic separation was performed on a Zorbax SB-AQ column (250 mm × 4.6 mm, 5 μm) at a wavelength of 220 nm using pH 4.0 phosphate buffer-acetonitrile as the mobile phase in gradient elution mode. The validation results demonstrate that the method has acceptable specificity, linearity, accuracy, precision and robustness. The detection limits and quantitation limits ranged from 2 to 17 ng/mL and from 6 to 50 ng/mL, respectively. A linear relationship was observed between the peak area and concentration of lenalidomide and impurities, with a correlation coefficient value of <em>r</em> ≥ 0.999. The formation and chemical structures of oxidation and base degradation products of lenalidomide have also been confirmed. The newly developed HPLC method is suitable for use in quality-control laboratories for qualitative and quantitative assessment of thirteen kinds of related substances in the lenalidomide API.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"260 ","pages":"Article 116823"},"PeriodicalIF":3.1,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687078","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}

引用次数: 0

In vitro metabolism and metabolite identification of eutylone using rat liver microsomes 利用大鼠肝脏微粒体对丁酮进行体外代谢和代谢物鉴定

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-17 DOI: 10.1016/j.jpba.2025.116827

Alexandre Barcia Godoi , Natalícia de Jesus Antunes , Leonardo Costalonga Rodrigues , Aline Franco Martins , José Luiz Costa

{"title":"In vitro metabolism and metabolite identification of eutylone using rat liver microsomes","authors":"Alexandre Barcia Godoi , Natalícia de Jesus Antunes , Leonardo Costalonga Rodrigues , Aline Franco Martins , José Luiz Costa","doi":"10.1016/j.jpba.2025.116827","DOIUrl":"10.1016/j.jpba.2025.116827","url":null,"abstract":"<div><div>The determination of toxicokinetic parameters allows the characterization of the toxicological profile of a wide range of substances. Several models have been employed to <em>in vitro</em> kinetic evaluations, in which liver microsomes stands out due to its simple handling and obtaining. Eutylone is a New Psychoactive Substance (NPS) classified as a synthetic cathinone with stimulant effects on central nervous system. This substance was initially synthesized for pharmaceutical applications but ultimately became subject to recreational use, with constant seizures worldwide. Herein, the <em>in vitro</em> metabolism and the production of eutylone phase I and phase II metabolites was investigated using rat liver microsomes (RLM). Eutylone presented a low metabolic stability showing an <em>in vitro</em> elimination half-life (t<sub>1/2</sub>) of 2.27 min. The unbound fractions of eutylone in microsomal (f<sub>u-mic</sub>) and plasma (f<sub>u-p</sub>) proteins were 0.93 and 0.15, respectively. A sigmoidal profile defined by Hill equation were observed, allowing kinetic parameters calculations. The Hill coefficient (H) was 1.21, <em>in vitro</em> maximum velocity (V<sub>max</sub>) was 19.40 μmol/mg/min, substrate concentration at half V<sub>max</sub> (S<sub>50</sub>) was 4.78 μM, intrinsic maximum clearance (CL<sub>max, in vitro</sub>) was 3.36 mL/min/mg, <em>in vivo</em> intrinsic clear-ance (CL<sub>int, in vivo</sub>) was 8.20 mL/min/kg, hepatic clearance (CL<sub>H</sub>) was 1.29 mL/min/kg, and hepatic extraction rate (E<sub>H</sub>) was 0.02. Eight eutylone metabolites were identified, four produced by phase I reactions and four by phase I followed by phase II reactions. Demethylenation and O-glucuronidation were pivotal in eutylone’s metabolism. These findings provide valuable information about the metabolism of eutylone, allowing practical implications for evaluating its safety and toxicity.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"260 ","pages":"Article 116827"},"PeriodicalIF":3.1,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687065","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}

引用次数: 0

Transitioning from concept to application: Comprehensive analytical validation of antibody-based smart sampling 从概念到应用的过渡：基于抗体的智能采样的全面分析验证

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-03-17 DOI: 10.1016/j.jpba.2025.116825

Ago Mrsa , Ridja Ishaq, Laila Brandoli, Trine Grønhaug Halvorsen , Léon Reubsaet

{"title":"Transitioning from concept to application: Comprehensive analytical validation of antibody-based smart sampling","authors":"Ago Mrsa , Ridja Ishaq, Laila Brandoli, Trine Grønhaug Halvorsen , Léon Reubsaet","doi":"10.1016/j.jpba.2025.116825","DOIUrl":"10.1016/j.jpba.2025.116825","url":null,"abstract":"<div><div>Smart sampling, introduced in 2018, is a technique that integrates crucial sample preparation steps for LC-MS protein analysis directly into the sampling process, improving efficiency. Currently most studies in this area are limited to proof of concept with a brief evaluation. Therefore, our objective is to fully validate the utility of smart samplers for quantitative analysis of bioactive proteins in biological matrices. This study builds on previous work where a paper-based antibody sampler was developed, and human chorionic gonadotropin (hCG) was used as a model protein. hCG is both a biomarker, a drug, and on the doping list for male athletes. Initially, the fabrication of the sampler was optimized, resulting in a reduction of antibody amount per sampler from 50 µg to 5 µg, reducing manufacturing cost while maintaining acceptable performance. It was also shown that removing the sampler before trypsination, but after reduction and alkylation greatly improved signal output while reducing the peptide background, suggesting a cleanser extract was achieved. The optimized smart sampler was validated for hCG in human serum using EMA’s ICH M10 guidelines on bioanalytical methods as a guide. Three calibration curves (seven levels) were made in the concentration range of 0.5 ng/mL to 75 ng/mL all displaying excellent linearity (r<sup>2</sup> ≥ 0.9955). The accuracy and precision both for the between-run (precision ≤ 10 % CV and accuracy 91–105 % (n = 3)) and within-run (precision ≤ 14 % CV and accuracy 94–105 % (n = 5)) runs were all well within the guideline's acceptance limits. The samplers also showed to be stable for at least ten weeks at room temperature. The optimization and validation of the antibody sampler detailed in this paper marks a notable progression, advancing the concept of \"smart sampling\" a step closer from the lab to practical application.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"260 ","pages":"Article 116825"},"PeriodicalIF":3.1,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687066","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}

引用次数: 0