Journal of Mass Spectrometry and Advances in the Clinical Lab最新文献

A liquid chromatography-high-resolution mass spectrometry method for separation and identification of hemoglobin variant subunits with mass shifts less than 1 Da

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-01-01 DOI: 10.1016/j.jmsacl.2025.01.002

Ainslie Chen , Ryan M. Aquino , Hector A. Vidal , Carolyn V. Wong , Ruben Y. Luo

Background

Identification of hemoglobin (Hb) variants is valuable in clinical testing. A common issue with conventional methods for identifying Hb variants is their subpar ability to provide structural breakdowns of the variants. Reports have surfaced of high-resolution mass spectrometry (HR-MS) methods that improve on traditional methods; however, ambiguities may arise without separation of Hb subunits prior to HR-MS analysis.

Methods

We report a liquid chromatography-high-resolution mass spectrometry (LC-HR-MS) method to separate several pairs of normal and variant Hb subunits with mass shifts of less than 1 Da and successfully identify them in intact-protein and top-down analyses. LC separation was facilitated by a C4 reversed-phase column.

Results

Seven heterozygous Hb variant samples (Hb C with α-thalassemia trait, Hb E, Hb D-Punjab, Hb G-Accra, Hb G-Siriraj, Hb Tarrant, and Hb G-Waimanalo) were selected to demonstrate the LC separation of Hb variant and normal subunits with mass shifts of less than 1 Da. The analytes could be explicitly observed in the deconvoluted MS¹ mass spectra. The top-down analysis matched the amino acid sequences of the correct Hb variant subunits.

Conclusions

The LC-HR-MS method described can effectively separate and identify Hb subunits, especially when the variant subunits have mass deviations of less than 1 Da from their corresponding normal subunits. With further evaluation to prove the clinical utility, the HR-MS methods including CE-HR-MS have the potential to complement or partially replace conventional methods of Hb variant identification in clinical laboratories.

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引用次数: 0

High-resolution mass spectrometry measurement of N-terminal carbamylated hemoglobin as a potential marker for chronic diseases with elevated blood urea levels

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-01-01 DOI: 10.1016/j.jmsacl.2025.01.001

Fangjun Chen , Priscilla S.-W. Yeung , Carolyn V. Wong , Ruben Y. Luo

Introduction

N-terminal carbamylated hemoglobin (CarHb) reflects long-term blood urea levels and has potential as a marker for chronic kidney disease (CKD) and other chronic conditions with elevated blood urea levels. A liquid chromatography-high-resolution mass spectrometry (LC-HR-MS) method was developed to measure CarHb.

Methods

Apparent CarHb/Hb ratios were calculated from the peak area ratios of carbamylated to native N-terminal peptides digested from hemoglobin alpha and beta subunits. Blood samples from healthy individuals, CKD patients, and chronic obstructive pulmonary disease (COPD) patients were analyzed.

Results

The apparent CarHb/Hb ratios were significantly higher in CKD and COPD patients compared to healthy individuals. However, no significant differences were observed between the CKD and COPD patient groups.

Conclusions

In this study, an LC-HR-MS method was developed for quantifying the apparent CarHb/Hb ratios and exploring their potential for clinical diagnostic applications. CarHb is a promising marker for monitoring kidney diseases and other chronic conditions with elevated blood urea levels. Beyond CarHb, the use of other carbamylated proteins as clinical diagnostic and prognostic markers can be explored.

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引用次数: 0

Development and clinical utility of an ultra performance liquid chromatography − tandem mass spectrometry assay for monitoring omadacycline and tigecycline in severe bacterial infections 开发超高效液相色谱-串联质谱测定法，用于监测严重细菌感染中的奥美沙星和替加环素，并提高其临床实用性

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2024-11-01 DOI: 10.1016/j.jmsacl.2024.11.001

Chang Wang , Bingfeng Luo , Wenqing Liu , Chen Jia , Haile Chen , Jingjing Ma , Xia Song , Xingfang Ji , Aijia Cao , Yinliang Bai , Wen Qiu

Objective

We aimed to develop a rapid, simple, and precise ultra performance liquid chromatography − tandem mass spectrometry (UPLC-MS/MS) technique for simultaneous measurement of omadacycline (OMA) and tigecycline (TGC) in the bloodstream of individuals suffering from serious bacterial infections.

Methods

All analytes were extracted using a 0.2 % formic acid–water dilution and acetonitrile plasma protein precipitation. The quantification was performed by electrospray ionization-triple quadrupole mass spectrometry with selected reaction monitoring and positive ion mode detection. Tetracycline was used as an internal standard in this experiment, with the mobile phase composed of water (with 0.1 % formic acid) and acetonitrile (using gradient elution) flowing at a rate of 0.35 ml/min, and the column temperature set at 30 °C. Each individual analysis was completed in under 3.5 min.

Results

The method was validated based on FDA recommendations, including the assessment of extraction recovery (92.65–101.72 %) and matrix effects (86.22–91.12 %). The standard curve ranges for both OMA and TGC are 0.025 µg/mL to 2.5 µg/mL. The plasma samples were found to be consistent after undergoing three rounds of freezing and thawing at room temperature for 24 h, being placed in an automated sample injector for 24 h, and then frozen for 45 days. Clinical cases were used to demonstrate the application of the therapeutic drug monitoring (TDM) assay, showing how an analytical test can quickly provide information on antibiotic levels in patients and impact their treatment.

Conclusion

Multiplex UPLC-MS/MS assays for the simultaneous measurement of plasma OMA and TGC concentrations are the ideal choice for clinically TDM applications.

目的我们旨在开发一种快速、简便、精确的超高效液相色谱-串联质谱（UPLC-MS/MS）技术，用于同时测定严重细菌感染患者血液中的奥美拉唑（OMA）和替加环素（TGC）。方法使用 0.2 % 甲酸-水稀释液和乙腈血浆蛋白沉淀提取所有分析物。采用电喷雾电离-三重四极杆质谱法进行定量，并进行选择反应监测和正离子模式检测。实验中使用四环素作为内标，流动相为水（含 0.1 % 甲酸）和乙腈（使用梯度洗脱），流速为 0.35 ml/min，柱温设定为 30 °C。结果根据 FDA 的建议对该方法进行了验证，包括评估萃取回收率（92.65%-101.72%）和基质效应（86.22%-91.12%）。OMA 和 TGC 的标准曲线范围为 0.025 µg/mL 至 2.5 µg/mL。血浆样本在室温下经过三轮冷冻和解冻 24 小时后，放入自动进样器中 24 小时，然后冷冻 45 天，结果一致。临床病例展示了治疗药物监测 (TDM) 分析的应用，说明了分析检验如何快速提供患者体内抗生素水平的信息，并对治疗产生影响。

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引用次数: 0

Tandem mass spectrometry of serum cholestanoic (C27) acids – Typical concentration ranges and application to the study of peroxisomal biogenesis disorders 血清胆甾烷酸（C27）的串联质谱法--典型浓度范围及在过氧化物酶体生物生成紊乱研究中的应用

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2024-11-01 DOI: 10.1016/j.jmsacl.2024.10.005

Wujuan Zhang , Monica Narvaez Rivas , Kenneth D.R. Setchell

Background

Primary bile acid synthesis is impaired in peroxisomal disorders, leading to the accumulation of long-chain bile acids, specifically dihydroxycholestanoic and trihydroxycholestanoic acids. Quantification of the diastereoisomers of these C₂₇ bile acids is essential for the differential diagnosis of these disorders.

Methods

High-performance liquid chromatography electrospray ionization-tandem mass spectrometry with stable-isotope dilution was used to quantify all eight diastereoisomers of cholestanoic acids in serum. Clinical ranges were established for patients with and without cholestatic liver disease, as well as for those with peroxisomal disorders.

Results

The assay was linear over the range of 20–2,500 ng/mL, and intra- and inter-assay imprecision was <20 % CV. The mean (±SEM) serum concentration of total C₂₇ bile acids in 20 adult controls was low (0.007 ± 0.004 μmol/L). In non-cholestatic, moderately cholestatic, and severely cholestatic patients, total C₂₇ bile acids measured 0.015 ± 0.011, 0.129 ± 0.034, and 0.986 ± 0.249 μmol/L, respectively. In contrast, patients with confirmed peroxisomal disorders (n = 49) exhibited concentrations >10-fold higher (14.06 ± 2.59 μmol/L). Patients with heterozygous mutations in PEX genes had low concentrations of serum C₂₇ bile acids. In all groups, the (25S)- and (25R)-diastereomers were present in a ratio of 0.3. In cases of 2-methylacyl-CoA racemase deficiency, serum total C₂₇ bile acids were markedly elevated (10.61 ± 0.92 μmol/L) and comprised exclusively the (25R)-diastereoisomer.

Conclusions

This tandem mass spectrometric assay quantifies all diastereoisomers of the C₂₇ cholestanoic acids in serum and was used to establish typical clinical concentration ranges. The method is applicable to the diagnosis of peroxisomal disorders and differentiates 2-methylacyl-CoA racemase deficiency from other peroxisomal biogenesis disorders.

背景过氧化物酶体失调时初级胆汁酸合成受损，导致长链胆汁酸积累，特别是二羟基胆甾烷酸和三羟基胆甾烷酸。方法采用高效液相色谱电喷雾离子化-串联质谱法和稳定同位素稀释法对血清中胆汁酸的所有八种非对映异构体进行定量分析。结果该测定在 20-2,500 纳克/毫升范围内呈线性关系，测定内和测定间的不精密度为 CV 值的 20%。20 名成人对照组血清中总 C27 胆汁酸的平均（±SEM）浓度较低（0.007 ± 0.004 μmol/L）。在非胆汁淤积型、中度胆汁淤积型和重度胆汁淤积型患者中，总 C27 胆汁酸的浓度分别为 0.015 ± 0.011、0.129 ± 0.034 和 0.986 ± 0.249 μmol/L。相比之下，确诊为过氧化物酶体紊乱的患者（n = 49）的浓度高出 10 倍（14.06 ± 2.59 μmol/L）。PEX基因发生杂合突变的患者血清中C27胆汁酸的浓度较低。在所有组别中，(25S)-和(25R)-非对映异构体的比例均为 0.3。在 2-甲基乙酰-CoA消旋酶缺乏症病例中，血清中 C27 胆汁酸总量明显升高（10.61 ± 0.92 μmol/L），且完全由 (25R)- 非对映异构体组成。该方法适用于过氧物酶体疾病的诊断，并能将2-甲基酰-CoA外消旋酶缺乏症与其他过氧物酶体生物生成疾病区分开来。

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引用次数: 0

Research letter: Pilot study of a prototype fully automated mass spectrometry system for routine clinical laboratory use 研究信：用于常规临床实验室的全自动质谱分析系统原型的试点研究

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2024-11-01 DOI: 10.1016/j.jmsacl.2024.10.004

Michael Vogeser , Katharina Habler , Antje Reuter , Christian Schneider-Thauern

引用次数: 0

Method validation of an inductively coupled plasma mass spectrometry (ICP-MS) assay for the analysis of magnesium, copper and zinc in red blood cells 电感耦合等离子体质谱法（ICP-MS）分析红细胞中镁、铜和锌的方法验证。

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2024-10-13 DOI: 10.1016/j.jmsacl.2024.10.003

Nazmin Bithi , Daniel Ricks , Brandon S. Walker , Christian Law , Kamisha L. Johnson-Davis

Background

Laboratory measurements of trace elements such as magnesium (Mg), copper (Cu), and zinc (Zn) in red blood cells (RBCs) are essential for assessing nutritional status and diagnosing metal toxicity. The purpose of this study was to develop and validate an ICP-MS method for quantifying these elements in RBCs.

Methods

Packed RBCs were aliquoted and diluted in an alkaline diluent solution containing internal standards, 0.1 % Triton X-100, 0.1 % EDTA, and 1 % ammonium hydroxide. The resulting diluted specimen was analyzed using inductively coupled plasma mass spectrometry (ICP-MS) to quantitatively determine the levels of Mg, Cu, and Zn. The method underwent validation for accuracy, precision, method comparison, linearity, analytical sensitivity, and carryover. Additionally, retrospective data were analyzed, and non-parametric reference intervals were calculated.

Results

Accuracy and linearity fell within the expected range of ≤±15 % for all analytes. Within-run, between-run, and total imprecision were ≤15 % coefficient of variation. All other validation experiments met the established acceptance criteria. Retrospective data analysis was conducted on patient samples using the method. The application of Tukey’s HSD test for multiple comparisons revealed statistically significant mean differences (p < 0.05) in Mg, Cu, and Zn concentrations between all pairwise groups of age and sex, except for the mean Cu concentration in adult males versus females and the mean Mg concentrations in adult versus minor males.

Conclusions

The presented method was successfully validated and met the criteria for clinical use. Retrospective data analysis of patient results demonstrated the method’s suitability for assessing nutritional deficiency and toxicity.

背景：红细胞（RBC）中镁（Mg）、铜（Cu）和锌（Zn）等微量元素的实验室测量对于评估营养状况和诊断金属中毒至关重要。本研究的目的是开发并验证一种 ICP-MS 方法，用于定量检测红细胞中的这些元素：方法：等分包装好的红细胞，用含有内标、0.1 % Triton X-100、0.1 % EDTA 和 1 % 氢氧化铵的碱性稀释液稀释。稀释后的样本使用电感耦合等离子体质谱法（ICP-MS）进行分析，以定量测定镁、铜和锌的含量。该方法经过了准确度、精密度、方法比较、线性度、分析灵敏度和携带率的验证。此外，还对回顾性数据进行了分析，并计算了非参数参考区间：结果：所有分析物的准确度和线性度均在预期范围内（≤±15%）。运行内、运行间和总不精密度的变异系数均小于 15%。所有其他验证实验均符合既定的验收标准。使用该方法对患者样本进行了回顾性数据分析。采用 Tukey's HSD 检验进行多重比较，结果显示平均差异有显著统计学意义（p 结论：该方法成功地验证了 "雌激素水平 "和 "雌激素水平"：该方法已成功通过验证，并符合临床使用标准。对患者结果的回顾性数据分析表明，该方法适用于评估营养缺乏和毒性。

{"title":"Method validation of an inductively coupled plasma mass spectrometry (ICP-MS) assay for the analysis of magnesium, copper and zinc in red blood cells","authors":"Nazmin Bithi , Daniel Ricks , Brandon S. Walker , Christian Law , Kamisha L. Johnson-Davis","doi":"10.1016/j.jmsacl.2024.10.003","DOIUrl":"10.1016/j.jmsacl.2024.10.003","url":null,"abstract":"<div><h3>Background</h3><div>Laboratory measurements of trace elements such as magnesium (Mg), copper (Cu), and zinc (Zn) in red blood cells (RBCs) are essential for assessing nutritional status and diagnosing metal toxicity. The purpose of this study was to develop and validate an ICP-MS method for quantifying these elements in RBCs.</div></div><div><h3>Methods</h3><div>Packed RBCs were aliquoted and diluted in an alkaline diluent solution containing internal standards, 0.1 % Triton X-100, 0.1 % EDTA, and 1 % ammonium hydroxide. The resulting diluted specimen was analyzed using inductively coupled plasma mass spectrometry (ICP-MS) to quantitatively determine the levels of Mg, Cu, and Zn. The method underwent validation for accuracy, precision, method comparison, linearity, analytical sensitivity, and carryover. Additionally, retrospective data were analyzed, and non-parametric reference intervals were calculated.</div></div><div><h3>Results</h3><div>Accuracy and linearity fell within the expected range of ≤±15 % for all analytes. Within-run, between-run, and total imprecision were ≤15 % coefficient of variation. All other validation experiments met the established acceptance criteria. Retrospective data analysis was conducted on patient samples using the method. The application of Tukey’s HSD test for multiple comparisons revealed statistically significant mean differences (p < 0.05) in Mg, Cu, and Zn concentrations between all pairwise groups of age and sex, except for the mean Cu concentration in adult males versus females and the mean Mg concentrations in adult versus minor males.</div></div><div><h3>Conclusions</h3><div>The presented method was successfully validated and met the criteria for clinical use. Retrospective data analysis of patient results demonstrated the method’s suitability for assessing nutritional deficiency and toxicity.</div></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"34 ","pages":"Pages 21-27"},"PeriodicalIF":3.1,"publicationDate":"2024-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11513474/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142523609","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Liquid chromatography-mass spectrometric method for the simultaneous analysis of branched-chain amino acids and their ketoacids from dried blood spot as secondary analytes for the detection of maple syrup urine disease 液相色谱-质谱法同时分析作为检测枫糖尿病辅助分析物的干血斑中支链氨基酸及其酮酸的方法

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2024-10-10 DOI: 10.1016/j.jmsacl.2024.10.001

Arya Raveendran , Ashutosh Gupta , Leslie E. Lewis , Krishnananda Prabhu , Sudheer Moorkoth

Background

Maple syrup urine disease (MSUD) is an aminoacidopathy caused by a defective branched-chain alpha-ketoacid dehydrogenase complex, leading to the accumulation of branched-chain amino acids (BCAAs) and their respective keto acids (BCKAs). A comprehensive test was developed to measure BCAAs and BCKAs using LC-MS from dried blood spot (DBS) samples for the diagnosis and prevention of MSUD in newborns and infants.

Methods

Analytes were extracted from DBS using a methanol:0.1 % v/v formic acid solution (75:25) containing internal standards and analyzed on a Luna PFP column (150 mm × 4.6 mm, 3 µm) at a flow rate of 0.3 mL/min. The method was validated for linearity, accuracy, precision, recovery, carry-over, matrix effect, hematocrit, blood volume, and punch position effects. Biomarker stability in the matrix and stock solution was assessed. Correlation with the plasma method was determined using Pearson’s correlation coefficient and Bland-Altman analysis. The method established reference ranges for the Udupi district population in South India.

Results

The method demonstrated linearity (r² > 0.99), with a lower limit of detection at 2 µM (BCAA) and 1 µM (BCKA), and acceptable recovery of QC samples. Hematocrit, blood volume, punch position, and storage condition effects were within acceptable limits. Correlation and Bland-Altman analysis showed strong interconvertibility between plasma and DBS assays. Reference ranges for leucine, isoleucine, valine, KIC, KIV, and KMV were established.

Conclusion

The developed DBS method, requiring no derivatization and involving simple sample preparation with short run times, is a cost-effective and reliable approach for the confirmatory diagnosis of MSUD.

背景枫糖浆尿病（MSUD）是由支链α-酮酸脱氢酶复合物缺陷引起的一种氨基酸病，导致支链氨基酸（BCAA）及其相应酮酸（BCKAs）的积累。方法用含内标物的甲醇：0.1 % v/v 甲酸溶液（75:25）从干血斑（DBS）样品中提取分析物，然后用 Luna PFP 色谱柱（150 mm × 4.6 mm, 3 µm）分析，流速为 0.3 mL/min。对该方法的线性、准确度、精密度、回收率、携带率、基质效应、血细胞比容、血容量和打孔位置效应进行了验证。评估了生物标记物在基质和储备溶液中的稳定性。采用皮尔逊相关系数和 Bland-Altman 分析法确定了与血浆法的相关性。结果该方法具有线性关系（r2 > 0.99），检测下限为 2 µM（BCAA）和 1 µM（BCKA），质控样品的回收率可接受。血细胞比容、血容量、打孔位置和储存条件的影响均在可接受的范围内。相关性和 Bland-Altman 分析表明血浆和 DBS 检测之间具有很强的相互转换性。结论：所开发的 DBS 方法无需衍生化，样品制备简单，运行时间短，是确诊 MSUD 的一种经济、可靠的方法。

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引用次数: 0

Development and validation of an LC-MS/MS method for highly concentrated tacrolimus and cyclosporine samples prepared from pharmaceutical products to assess drug loss from feeding tubes 针对从药品中制备的高浓度他克莫司和环孢素样品开发和验证 LC-MS/MS 方法，以评估输液管中的药物流失情况

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2024-10-09 DOI: 10.1016/j.jmsacl.2024.10.002

Yi Xiao , Mari Ishak Gabra , Edward Leung

Introduction

Tacrolimus and cyclosporine are common immunosuppressants utilized post-organ transplantation to manage allograft rejection. Both have narrow therapeutic indices and are frequently measured to support dose adjustments. Although nasogastric tubes are commonly used to provide nutritional support and serve as a route for immunosuppressant administration, they were never validated for such purposes.

Objective

To develop and validate a liquid chromatography – tandem mass spectrometry (LC-MS/MS) method for highly concentrated tacrolimus and cyclosporine samples prepared from pharmaceutical products to support the validation of feeding tube administration of these immunosuppressants.

Methods

The method involved stepwise dilutions with dimethyl sulfoxide before analysis using online sample preparation and LC-MS/MS. It was validated in a CLIA-certified clinical laboratory that measures immunosuppressants by LC-MS/MS and is designed to support clinical studies evaluating drug loss from feeding tubes.

Results

The method was linear between 6.8 µg/mL and 75 µg/mL for tacrolimus, and between 0.9 mg/mL and 10 mg/mL for cyclosporine, with r² > 0.99 and total precision <5 % at all QC levels. The method demonstrated good recovery using cyclosporine Certified Reference Material, tacrolimus European Pharmacopeia Reference Standard, and prepared pharmaceutical products. Minimal matrix effects were observed.

Conclusion

An analytical method was developed and validated for in vitro studies with simulated administration of tacrolimus or cyclosporine to assess loss during drug administration using feeding tubes.

导言他克莫司和环孢素是器官移植后用于控制异体移植排斥反应的常用免疫抑制剂。这两种药物的治疗指数都很窄，需要经常测量以支持剂量调整。虽然鼻胃管通常用于提供营养支持并作为免疫抑制剂的给药途径，但它们从未被验证用于此类目的。目标开发并验证一种液相色谱-串联质谱（LC-MS/MS）方法，用于检测从药品中制备的高浓度他克莫司和环孢素样品，以支持这些免疫抑制剂的喂管给药验证。结果该方法在他克莫司6.8微克/毫升至75微克/毫升、环孢素0.9毫克/毫升至10毫克/毫升之间呈线性关系，在所有质控水平下，r2为0.99，总精密度为5%。使用环孢素标准物质、他克莫司欧洲药典标准物质和制备的药品，该方法的回收率良好。结论开发并验证了一种分析方法，该方法适用于他克莫司或环孢素模拟给药的体外研究，以评估使用输液管给药过程中的药物损失。

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引用次数: 0

Rapid and sensitive liquid chromatographic–tandem mass spectrometric methods for the quantitation of dolutegravir in human plasma and breast milk 快速灵敏的液相色谱-串联质谱法定量检测人血浆和母乳中的多鲁曲韦

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2024-09-19 DOI: 10.1016/j.jmsacl.2024.09.001

Ashley R. Rackow , Aashish Pandey , Amelia L. Price , Mark A. Marzinke

Background

Dolutegravir (DTG) is part of a first-line antiretroviral therapy (ART) for HIV management in drug-naïve individuals and is recommended for the treatment of HIV during pregnancy. Robust analytical tools to quantify DTG are necessary to support clinical trials that characterize its multi-compartment drug distribution.

Methods

Potassium EDTA (K₂EDTA) plasma or whole breast milk was spiked with DTG and an isotopically labeled internal standard. Samples were prepared via protein precipitation prior to LC–MS/MS analysis. The assays were validated in accordance with regulatory recommendations.

Results

Analytical measuring ranges for DTG quantitation in plasma and breast milk were 100–10,000 ng/mL and 0.500 to 1000 ng/mL, respectively. Inter-assay precision and accuracy were 2.73 % to 3.41 % and −10.6 % to −5.37 % for plasma, and 4.24 % to 12.4 % and −5.63 % to 7.49 % for breast milk, respectively. DTG was stable for three freeze–thaw cycles and for at least 72 h at room temperature in matrix (plasma or breast milk). Additionally, whole blood was stable for 24 h at room temperature and 2 h under conditions of extended heat and humidity. Matrix effects for DTG in plasma and breast milk ranged from 101 % to 108 % and 78.2 % to 99.3 %, respectively. Quantitation in remnant plasma samples yielded measurable concentrations within the primary linearity of the assay.

Conclusions

Methods to quantify DTG in human plasma and breast milk have been developed and validated. These assays were designed to satisfy all criteria for implementation in clinical and clinical trial settings.

背景Dolutegravir（DTG）是一线抗逆转录病毒疗法（ART）的一部分，用于治疗药物过敏者的艾滋病，并被推荐用于妊娠期艾滋病的治疗。方法在乙二胺四乙酸钾（K2EDTA）血浆或全母乳中添加 DTG 和同位素标记的内标。在进行 LC-MS/MS 分析之前，通过蛋白质沉淀法制备样品。结果血浆和母乳中 DTG 定量的分析测量范围分别为 100-10,000 纳克/毫升和 0.500-1000 纳克/毫升。血浆的测定间精密度和准确度分别为 2.73 % 至 3.41 % 和 -10.6 % 至 -5.37 %，母乳的测定间精密度和准确度分别为 4.24 % 至 12.4 % 和 -5.63 % 至 7.49 %。在基质（血浆或母乳）中，DTG 在三次冻融循环和室温下至少 72 小时内都是稳定的。此外，全血在室温下稳定 24 小时，在高温高湿条件下稳定 2 小时。血浆和母乳中 DTG 的基质效应分别为 101 % 至 108 % 和 78.2 % 至 99.3 %。残余血浆样本中的定量结果显示，可测量的浓度在检测方法的主要线性范围内。这些检测方法的设计符合在临床和临床试验环境中实施的所有标准。

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引用次数: 0

Understanding isotopes, isomers, and isobars in mass spectrometry 了解质谱中的同位素、同分异构体和同分异构体

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2024-08-01 DOI: 10.1016/j.jmsacl.2024.08.002

Katharina Habler, Arber Rexhaj, Manuela Adling-Ehrhardt, Michael Vogeser

Mass spectrometry (MS) is a versatile analytical tool used in various fields such as biochemistry, pharmacology, omics, and clinical analysis for determining and quantifying compounds based on their molecular mass and structure through the mass-to-charge ratio. While MS offers high specificity and selectivity, it encounters challenges including matrix effects, in-source fragmentation, and other interferences caused by natural isotopic abundance, as well as isomeric and isobaric compounds. These challenges can impede accurate qualitative and quantitative analysis. Visual aids such as graphical illustrations can help elucidate the chemical differences and similarities among isotopes, isomers, and isobaric compounds.

质谱（MS）是一种多功能分析工具，可用于生物化学、药理学、组学和临床分析等多个领域，通过质量电荷比，根据化合物的分子质量和结构对其进行测定和量化。虽然质谱具有高度的特异性和选择性，但它也会遇到各种挑战，包括基质效应、源内碎裂、由天然同位素丰度以及同分异构体和同位化合物引起的其他干扰。这些挑战会妨碍准确的定性和定量分析。图解等可视化辅助工具有助于阐明同位素、同分异构体和等压化合物之间的化学异同。

引用次数: 0