Journal of Chromatography B最新文献

{"title":"LC-MS analysis of chiral amino acids in human urine reveals D-amino acids as potential biomarkers for colorectal cancer","authors":"Wenchan Deng ,&nbsp;Chundan Ye ,&nbsp;Wei Wang ,&nbsp;Rongrong Huang ,&nbsp;Cheng Guo ,&nbsp;Yuanjiang Pan ,&nbsp;Cuirong Sun","doi":"10.1016/j.jchromb.2024.124270","DOIUrl":"10.1016/j.jchromb.2024.124270","url":null,"abstract":"<div><p>Colorectal cancer (CRC) is a common malignant tumor in the gastrointestinal tract. Changes in amino acid metabolites have been implicated in tumorigenesis and disease progression. Biomarkers on the basis of chiral amino acids, especially D-amino acids, have not been established for early diagnosis of CRC. Quantification of chiral amino acids, especially very low concentrations of endogenous D-amino acids, is technically challenging. We report here the quantification of L- and D-amino acids in urine samples collected from 115 CRC patients and 155 healthy volunteers, using an improved method. The method of chiral labeling, liquid chromatography, and tandem mass spectrometry enabled separation and detection of 28 amino acids (14 L-amino acids, 13 D-amino acids and Gly). Orthogonal partial least squares discriminant analysis identified 14 targeted variables among these chiral amino acids that distinguished the CRC from the healthy controls. Binary logistic regression analysis revealed that D-α-aminobutyric acid (D-AABA), L-alanine (L-Ala), D-alanine (D-Ala), D-glutamine (D-Gln) and D-serine (D-Ser) could be potential biomarkers for CRC. A receiver operating characteristic curve analysis of combined multi-variables contributed to an area under the curve (AUC) of 0.995 with 98.3 % sensitivity and 96.8 % specificity. A model constructed with D-AABA, D-Ala, D-Gln, and D-Ser achieved an AUC of 0.988, indicating important contributions of D-amino acids to the association with CRC. Further analysis also demonstrated that the metabolic aberration was associated with age and the development of CRC, D-methionine (D-Met) was decreased in CRC patients with age over 50, and D/L-Gln in patients at stage IV was higher than patients at stage I. This study provides the signature of D-amino acids in urine samples and offers a promising strategy for developing non-invasive diagnosis of CRC.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1245 ","pages":"Article 124270"},"PeriodicalIF":2.8,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141911142","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":"Evaluation of pharmacokinetic herb-drug interaction of diabecon and losartan by UHPLC-MS/MS","authors":"V.V.S. Prasanna Kumari Rayala,&nbsp;Rushikesh Shantilal Shinde,&nbsp;Abhay Kumar,&nbsp;P. Radhakrishnanand","doi":"10.1016/j.jchromb.2024.124267","DOIUrl":"10.1016/j.jchromb.2024.124267","url":null,"abstract":"<div><p>The diabecon is an ayurvedic herbal formulation that contains a mixture of herbs traditionally used as antidiabetic which is reported in the ayurvedic pharmacopeia of India and Indian Materia medica. The diabetic population has a common co-morbidity of hypertension for which losartan drug is commonly used for the treatment of hypertension. However, there is a lack of research on the pharmacokinetics interaction between diabecon and losartan. This research aims to investigate the influence of diabecon on the pharmacokinetics of losartan drugs in rats by establishing a highly sensitive ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry (UHPLC-MS/MS) method. The method was validated according to the USFDA guidelines and was applied for the pharmacokinetic study. The lowest concentration of losartan detection in rat plasma was found to be 1 ng/mL and the accuracy and precision were within the linear range (1–1500 ng/mL). The results revealed that after 28 days of dosing diabecon, it has altered the pharmacokinetic parameters like area under the curve (AUC_0-t), drug clearance (Cl/F), and rate of elimination (Ke) of losartan, which may happen due to enzyme induction. Although there was a change in the pharmacokinetic parameters of losartan when administered in the presence of diabecon, it was found to be nonsignificant in rats (p &gt; 0.05). According to the best of our knowledge, this is the first article that discusses the pharmacokinetic herb-drug interaction between diabecon and losartan.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1245 ","pages":"Article 124267"},"PeriodicalIF":2.8,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141911140","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":"Innovative disposable in-tip cellulose paper (DICP) device for facile determination of pesticides in postmortem blood samples: A proof-of-concept study","authors":"Rajeev Jain ,&nbsp;Mukesh Kumar Singh ,&nbsp;Nemat Ali ,&nbsp;Mohammad Rashid Khan ,&nbsp;Atul Bajaj ,&nbsp;Mohana Krishna Reddy Mudiam","doi":"10.1016/j.jchromb.2024.124268","DOIUrl":"10.1016/j.jchromb.2024.124268","url":null,"abstract":"<div><p>Accurately identifying and quantifying toxicants is crucial for medico-legal investigations in forensic toxicology; however, low analyte concentrations and the complex samples matrix make this work difficult. Therefore, a simplified sample preparation procedure is crucial to streamline the analysis to minimize sample handling errors, reduce cost and improve the overall efficiency of analysis of toxicants. To address these challenges, an innovative disposable in-tip cellulose paper (DICP) device has been developed for the extraction of three pesticides viz. Chlorpyrifos, Quinalphos and Carbofuran from postmortem blood samples. The DICP device leverages cellulose paper strips housed within a pipette tip to streamline the extraction process, significantly reducing solvent usage, time, and labor while maintaining high analytical accuracy. The extraction of pesticides from postmortem blood using the DICP device involves a streamlined process characterized by adsorption and desorption. The diluted blood samples were processed through the DICP device via repeated aspirating and dispensing calyces to adsorb the pesticides onto the cellulose paper. The adsorbed pesticides are then eluted using acetone, which is collected for GC–MS analysis. The method was meticulously optimized, achieving a limit of quantification in the range of 0.009–0.01 µg mL⁻¹. The intra-day and inter-day precisions were consistently less than 5 % and 10 %, respectively, with accuracy ranging from 94–106 %. Relative recoveries for the analytes were observed to be between 60 % and 93.3 %, and matrix effects were determined to be less than 10 %. The method’s sustainability was validated with a whiteness score of 98.8, an AGREE score of 0.64, a BAGI score of 70 and ComplexMoGAPI score of 77. Applicability was demonstrated through successful analysis of real postmortem blood samples and proficiency testing samples, highlighting its potential utility in forensic toxicology.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1245 ","pages":"Article 124268"},"PeriodicalIF":2.8,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141911141","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":"Volatile and non-volatile metabolite profiling of under-explored crop Piper colubrinum Link. with chromatography mass spectrometry approach and its biochemical diversity from medicinally valued Piper species","authors":"D. Sruthi ,&nbsp;T. John Zachariah","doi":"10.1016/j.jchromb.2024.124260","DOIUrl":"10.1016/j.jchromb.2024.124260","url":null,"abstract":"<div><p><em>Piper colubrinum</em> Link. is an underexplored crop regarding its metabolites and therapeutic attributes. Current study aimed to identify the possible volatile and non-volatile metabolites of <em>P. colubrinum</em> fruit and studied its metabolite diversity with medicinally valued <em>Piper</em> species <em>viz. P. nigrum</em> L.<em>, P. longum</em> L. and <em>P. chaba</em> Hunter. The volatile constituents of <em>P. colubrinum</em> essential oil by GC–MS revealed the presence of sesquiterpenes as the major contribution. The sesquiterpenes α-muurolol (12.5 %) and β-caryophyllene (11.3 %) were the predominant volatile components. Few aliphatic compounds like n-heptadecane and trace amounts of monoterpenes (α- and β-pinene and α-terpineol) were also identified from this crop. The fatty acid profiling by GC–MS revealed mainly oleic acid (41.3 %) followed by palmitic and linoleic acids. HPLC analysis demonstrated that the major pungent alkaloid piperine was found to be trace (0.04 %) in <em>P. colubrinum</em>. The LC-QTOF-MS/MS profiling of the chloroform extract of the <em>P. colubrinum</em> revealed the presence of non-volatile constituents including phenolic and alkaloid compounds. Ferulic acid, rosmarinic acid, salicylic acid, kaempferol-5-glucoside, 5-methoxysalicylic acid, apigenin-7-galactoside, kaempferide-3-glucoside, luteolin, kaempferol, apigenin and scutellarein-4′-methyl ether were the phenolic compounds whereas piperlonguminine was the alkaloid compound identified. Finally, the biochemical parameters of this crop were compared with that of <em>P. nigrum, P. longum</em> and <em>P. chaba</em> and average linkage cluster dendrogram revealed that <em>P. colubrinum</em> was biochemically distinct from other three <em>Piper</em> species.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1245 ","pages":"Article 124260"},"PeriodicalIF":2.8,"publicationDate":"2024-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142040813","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":"Digital identification and adulteration analysis of Pulsatilla Radix and Pulsatilla Cernua based on “digital identity” and UHPLC-QTOF-MSE","authors":"Xian rui Wang ,&nbsp;Jia ting Zhang ,&nbsp;Wen guang Jing ,&nbsp;Ming hua Li ,&nbsp;Xiao han Guo ,&nbsp;Xian long Cheng ,&nbsp;Feng Wei","doi":"10.1016/j.jchromb.2024.124257","DOIUrl":"10.1016/j.jchromb.2024.124257","url":null,"abstract":"<div><p>Under the background of digitalization of traditional Chinese medicine (TCM), to realize the quick identification and adulteration analysis of <em>Pulsatilla Radix</em> (PR), adhering to digital conviction, this study conducted UHPLC-QTOF-MS^E analysis on PR and its adulterant-<em>Pulsatilla Cernua</em> (PC) from different batches and based on digital conversion, the shared ions were extracted from different batches of PR and PC as their “ions representation”, respectively. Further, the data set of unique ions of PR relative to PC and PC relative to PR were screened out as the “digital identities” of PR and PC respectively. Further, above the “digital identities” of PR and PC were used as the benchmarks for matching and identifying to feedback give a matching credibility (MC). The results showed that based on the “digital identities” of PR and PC, the digital identification of two herbal samples can be realized efficiently and accurately at the individual level with the MC≥70.00 %, even if 5 % of PC in the mixed samples can still be identified efficiently and accurately. The study is of great practical significance for improving the identification efficiency of PR and PC, cracking down on adulterated and counterfeit drugs, and strengthening the quality control of PR. In addition, it has important reference significance for developing non-targeted digital identification of herbal medicines at the individual level based on UHPLC-QTOF-MS^E and the “digital identity”, which was beneficial to the construction of digital Chinese medicine and digital quality control.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1244 ","pages":"Article 124257"},"PeriodicalIF":2.8,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141873789","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":"A sensitive, expandable AQC-based LC-MS/MS method to measure amino metabolites and sphingolipids in cell and serum samples","authors":"Xiaotian Li ,&nbsp;Shuwei Tian ,&nbsp;Isabelle Riezman ,&nbsp;Yujiao Qin ,&nbsp;Howard Riezman ,&nbsp;Suihan Feng","doi":"10.1016/j.jchromb.2024.124256","DOIUrl":"10.1016/j.jchromb.2024.124256","url":null,"abstract":"<div><p>Sphingolipids are a major lipid species found in all eukaryotes. Among structurally complex and diversified lipids, sphingoid bases have been heavily linked to various metabolic diseases. However, most current LC-MS-based methods lack the sensitivity to detect low-abundant sphingoid bases. The 6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) derivatization reagent, which efficiently forms covalent bonds with amino groups, has been widely used for amino acid detection. Nevertheless, the commonly used reverse-phase HPLC method for amino acid analysis is not suitable for amphipathic sphingolipids. To address this issue, we report a robust reverse-phase HPLC-MS/MS method capable of separating and detecting hydrophilic amino acids and sphingoid bases in a single run with high sensitivity. This method is also inclusive of other amino metabolites with an expandable target list. We tested this method under various conditions and samples, demonstrating its high reproducibility and sensitivity. Using this approach, we systematically analyzed human serum samples from healthy individuals, dyslipidemia, and type II diabetes mellitus (T2DM) patients, respectively. Two sphingolipids and five amino acids were identified with significant differences between the control and T2DM groups, highlighting the potential of this method in clinical studies.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1245 ","pages":"Article 124256"},"PeriodicalIF":2.8,"publicationDate":"2024-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141846087","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":"Characterization of chemical constituents and metabolites in rat plasma after oral administration of Ainsliaea fragrans Champ by using UHPLC-QTOF-MS/MS","authors":"Zhaojun Wang ,&nbsp;Min Yao ,&nbsp;Hui Ouyang,&nbsp;Mingzhen He,&nbsp;Wentong Zhao,&nbsp;Wei Wei,&nbsp;Yushun Cui,&nbsp;Shilin Yang,&nbsp;Guoyue Zhong,&nbsp;Yulin Feng,&nbsp;Junmao Li","doi":"10.1016/j.jchromb.2024.124259","DOIUrl":"10.1016/j.jchromb.2024.124259","url":null,"abstract":"<div><p><em>Ainsliaea fragrans</em> Champ, a strong heat-clearing and detoxifying traditional Chinese medicine, has been effectively used for treating chronic cervicitis, endometritis, pelvic inflammatory diseases, and other conditions caused by damp heat. It shows a good effect in the treatment of cervicitis and has broad clinical application prospects. Nevertheless, there is no comprehensive study on its in vivo and in vitro chemical analysis. UHPLC-QTOF-MS/MS combined with the non-targeted characteristic filter analysis were used to conjecture and characterize the chemical components and in vivo metabolites of rats following oral administration of <em>Ainsliaea fragrans</em> Champ. In this study, A total of 85 compounds were identified in <em>Ainsliaea fragrans</em> Champ, including 29 flavonoids, 14 sesquiterpenoids, 25 chlorogenic acids, and 17 other compounds. In the plasma of rats after administration of <em>Ainsliaea fragrans</em> Champ, 160 compounds were deduced (19 prototype compounds and 141 metabolites). The 141 metabolites consist of 50 flavonoids, 80 phenolic acids and 11 Chlorogenic acids. The related metabolic pathways mainly involved demethylation, reduction, sulfonation, decarboxylation, hydroxylation, methylation, and glucuronide conjunction. In summary, the chemical components and metabolites of <em>Ainsliaea fragrans</em> Champ were comprehensively identified by using a rapid and accurate analysis method, which laid a foundation for dissecting its bioactive substances. In addition, it provides a scientific basis for the in-depth study of the material basis of <em>Ainsliaea fragrans</em> Champ efficacy and theoretical support for illustrating the mechanism of medical action and its clinical application.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1244 ","pages":"Article 124259"},"PeriodicalIF":2.8,"publicationDate":"2024-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141839591","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":"Membrane chromatography for AAV full capsid enrichment: Process development to scale up","authors":"Julio Huato Hernandez,&nbsp;Kurt Boenning,&nbsp;Aydin Kavara,&nbsp;Mark Schofield","doi":"10.1016/j.jchromb.2024.124258","DOIUrl":"10.1016/j.jchromb.2024.124258","url":null,"abstract":"<div><p>The recent FDA approval of several adeno-associated virus (AAV)-based gene therapies is driving demand for AAV production. One of the biggest AAV manufacturing challenges is removing “empty” capsids, which do not contain the gene of interest. Anion exchange chromatography has emerged as the leading solution for scalable full capsid enrichment. Here we develop a process for the baseline separation of empty and full AAV capsids using anion exchange membrane chromatography. This process development approach utilized AAV serotypes 8 and 9 and traverses initial screening of separation conditions up to manufacturing-scale processes. Process development of a two-step elution was performed via response surface DoE, exploring conductivity and the length of the first elution step. The results from response surfaces were used to construct statistical models of the process operating space. These models provide optimal conditions for recovery and purity, both of which can exceed 70 %. Model predictions were then validated at small scale prior to scale-up. We present the results from our scale-up purification and show that purity and yield are consistent with the results obtained from the response surface model.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1244 ","pages":"Article 124258"},"PeriodicalIF":2.8,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141851873","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":"Development and application of UPLC-Q-Orbitrap HRMS methods for the determination of eight metabolites of p-chloronitrobenzene in human urine","authors":"Peng Wang ,&nbsp;Qiuliang Xu ,&nbsp;Hua Zou ,&nbsp;Ming Liu ,&nbsp;Xiangjing Gao ,&nbsp;Hong Ren ,&nbsp;Junlin Wang ,&nbsp;Yiyao Cao","doi":"10.1016/j.jchromb.2024.124254","DOIUrl":"10.1016/j.jchromb.2024.124254","url":null,"abstract":"<div><p>Accurate, reliable, and sensitive methods for the determination of eight metabolites of p-chloronitrobenzene (p-CNB) were developed based on ultra-performance liquid chromatography − quadrupole − orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS). The free and conjugated forms of metabolites were determined before and after urine samples were hydrolyzed with acid. Subsequently, three solid phase extraction steps were used for concentration and purification. The calibration curves of the eight metabolites exhibited good linearity with an R² of &gt;0.999, and the precision was good as well, with the coefficient of variations of intra-day and inter-day being lower than 7.0 % and 8.5 %, respectively. Analytical accuracy for all metabolites varied within ranges of 76.0–102.9 %, and the limit of detection and limit of quantification of all the metabolites varied within ranges of 0.2–7.7 μg/L and 0.6–25.6 μg/L in human urine, respectively. In addition, the application potential of the proposed methods were evaluated by applying them to the determination of metabolites in the urine of workers exposed to p-CNB, and these results showed that these methods were accurate, reliable, and sensitive, which makes them an excellent choice for detecting the metabolites of p-CNB in the urine of exposed workers.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1244 ","pages":"Article 124254"},"PeriodicalIF":2.8,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141776150","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":"Simultaneous determination of total homocysteine, methionine, methylmalonic acid and 2-methylcitric acid in dried blood spots by ultra-performance liquid chromatography–tandem mass spectrometry","authors":"Zhenzhen Hu ,&nbsp;Lingwei Hu ,&nbsp;Chao Zhang ,&nbsp;Xiaoshan Yin ,&nbsp;Yu Zhang ,&nbsp;Kexin Fang ,&nbsp;Benqing Wu ,&nbsp;Xinwen Huang","doi":"10.1016/j.jchromb.2024.124253","DOIUrl":"10.1016/j.jchromb.2024.124253","url":null,"abstract":"<div><p>Homocysteine, methionine, methylmalonic acid and 2-methylcitric acid are clinically relevant markers in the methionine, propionate, and cobalamin metabolism. This study aimed to develop and validate an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneously determining total homocysteine, methionine, methylmalonic acid and 2-methylcitric acid in dried blood spots. Three 3.2 mm discs were punched from each calibrator, quality control, and sample dried blood spot into a 96-well U-plate. Each sample was spiked with internal standards and extracted. Then the supernatant was transferred to another 96-well U-plate. After nitrogen drying, the dried residues were reconstituted, centrifuged, and the resulting supernatant was transferred to another 96-well plate for analysis. The method was performed using UPLC-MS/MS within 3 min, validated according to guidance documents, and applied to 72 samples from confirmed patients with methionine, propionate, and cobalamin metabolism disorders. The UPLC-MS/MS method provided satisfactory separation of the four analytes. The R² values were ≥ 0.9937 for all analytes. The recoveries ranged from 94.17 to 114.29 %, and the coefficients of variation for intraday and interday precision were 0.19 % to 5.23 % and 1.02 % to 6.89 %, respectively. No significant carry-over was detected for the four analytes, and most of confirmed samples exhibited biomarker patterns characteristic of the relevant disorders. A simple and fast UPLC-MS/MS method was successfully developed, validated, and applied to clinical samples for the simultaneous determination of total homocysteine, methionine, methylmalonic acid, and 2-methylcitric acid in dried blood spots.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1244 ","pages":"Article 124253"},"PeriodicalIF":2.8,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141776151","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}