Journal of Mass Spectrometry and Advances in the Clinical Lab最新文献_第3页

Investigation of potential biomarkers for psoriasis in skin with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and ambient ionization mass spectrometry 基质辅助激光解吸/电离飞行时间质谱法和环境电离质谱法研究皮肤银屑病的潜在生物标志物

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-04-01 DOI: 10.1016/j.jmsacl.2025.04.004

Yi-Wen Hsu , Hung Su , Deng-Chyang Wu , Chi-Wei Lee , Sung-Jen Hung , Jentaie Shiea

Background

Psoriasis is a chronic inflammatory disease with an unclear etiology that affects skin, nails, and joints and often accompanies comorbidities. Recent studies indicate that alterations in metabolites within psoriatic lesions might be linked to inflammation. Studying bioactive lipid mediators or metabolites in skin inflammation and immunity might provide new potential biomarkers and therapeutic prediction factors.

Methods

Lipids and peptides in the scale extracts from psoriasis patients and healthy controls were characterized by thermal desorption-electrospray ionizationmass spectrometry and matrix-assisted laser desorption/ionization time-of-flightmass spectrometry, respectively. Principal component analysis (PCA) was then applied to these data to identify potential differences between psoriasis patients and healthy controls.

Results

Psoriatic plaques show reduced wax esters and triglycerides and a predominant increase in human neutrophil defensins (HNPs), compared to non-lesional sites of psoriatic patients and healthy control. Additionally, when medical treatments were administered to psoriasis patients, levels of HNPs were significantly reduced, suggesting that they may serve as potential biomarkers to evaluate therapeutic efficacy for psoriasis.

Conclusion

Two mass spectrometric techniques were used to rapidly and non-invasively identify and monitor potential biomarkers between psoriasis patients and healthy controls. However, PCA results only showed slight differences, and we intend to broaden the sample base in the future to increase the statistical power of the investigation.

背景银屑病是一种慢性炎症性疾病，病因不明，可累及皮肤、指甲和关节，常伴有合并症。最近的研究表明，银屑病病变内代谢物的改变可能与炎症有关。研究皮肤炎症和免疫中的生物活性脂质介质或代谢物可能提供新的潜在生物标志物和治疗预测因子。方法采用热解吸-电喷雾电离质谱法和基质辅助激光解吸/电离飞行时间质谱法分别对银屑病患者和健康对照鳞片提取物中的胶体和多肽进行表征。然后将主成分分析（PCA）应用于这些数据，以确定牛皮癣患者与健康对照之间的潜在差异。结果与银屑病患者和健康对照组相比，银屑病斑块显示蜡酯和甘油三酯减少，人中性粒细胞防御素（HNPs）明显增加。此外，当银屑病患者接受药物治疗时，HNPs水平显着降低，这表明它们可能作为评估银屑病治疗效果的潜在生物标志物。结论两种质谱技术可快速、无创地鉴定和监测银屑病患者与健康对照者之间的潜在生物标志物。然而，PCA结果只显示出轻微的差异，我们打算在未来扩大样本基础，以增加调查的统计能力。

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引用次数: 0

Quantification of glucagon and oxyntomodulin by protein precipitation-immunoaffinity enrichment-LC-MS/MS 蛋白沉淀-免疫亲和富集- lc -MS/MS定量胰高血糖素和氧合调节素

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-04-01 DOI: 10.1016/j.jmsacl.2025.04.002

Jessica O. Becker , Sara K. Shijo , Huu-Hien Huynh , Katrina L. Forrest , Michael J. MacCoss , Michelle A. Emrick , Elisha Goonatilleke , Andrew N. Hoofnagle

Introduction

Glucagon and oxyntomodulin are peptide hormones differentially released from proglucagon that function in regulating blood glucose. Their overlapping amino acid sequences make the development of specific immunoassays difficult, but the specificity of liquid chromatography-tandem mass spectrometry can be used to distinguish the peptides. We aimed to develop a sensitive and specific mass spectrometric assay that uses non-proprietary reagents and normal-flow liquid chromatography in the simultaneous quantification of both analytes.

Methods

Bulk plasma proteins were precipitated in ethanol/ammonium hydroxide. Analytes were enriched using monoclonal antibodies generated in-house and analyzed using liquid chromatography-tandem mass spectrometry. A glucagon calibration material was sourced commercially and characterized for purity and concentration by high-performance liquid chromatography-ultraviolet detection and amino acid analysis. Single-point calibration was used to minimize between-day variability.

Results

The novel antibodies performed acceptably (peptide recovery 45–59 %). The assay was precise (<13 %CV) and linear over the range of 1.3–14.7 pM and 1.1–13.7 pM for glucagon and oxyntomodulin, respectively. The glucagon calibration material concentration was determined to be 1.596 mg/g. Tube-type studies supported the use of protease inhibitor tubes at the time of blood draw. Patients with type 1 diabetes had lower concentrations of glucagon when maintained on an insulin pump, but not with injectable insulin.

Conclusion

We have validated a method with a highly detailed standard operating procedure. We have characterized calibration materials to help maintain accuracy and achieve between-day and between-laboratory harmonization. The assay will be beneficial in better understanding α-cell health and glycemic control in diabetes and other diseases.

胰高血糖素和氧合调节素是胰高血糖素原释放的肽激素，具有调节血糖的作用。它们重叠的氨基酸序列使特异性免疫测定的发展变得困难，但液相色谱-串联质谱法的特异性可以用来区分肽。我们的目标是开发一种敏感和特定的质谱分析方法，该方法使用非专有试剂和正常流动液相色谱法同时定量分析这两种分析物。方法用乙醇/氢氧化铵沉淀血浆蛋白。分析物使用内部生成的单克隆抗体进行富集，并使用液相色谱-串联质谱法进行分析。通过高效液相色谱-紫外检测和氨基酸分析对胰高血糖素的纯度和浓度进行了表征。采用单点校准以尽量减少日之间的变化。结果新抗体的多肽回收率为45 ~ 59%，表现良好。测定胰高血糖素和氧同调素的准确度（13% CV）在1.3-14.7 pM和1.1-13.7 pM范围内呈线性。测定胰高血糖素校准物质浓度为1.596 mg/g。管型研究支持在抽血时使用蛋白酶抑制剂管。1型糖尿病患者维持胰岛素泵时胰高血糖素浓度较低，而注射胰岛素时则没有。结论本方法具有非常详细的标准操作规程。我们对校准材料进行了表征，以帮助保持准确性并实现日与实验室之间的协调。该试验将有助于更好地了解糖尿病和其他疾病的α-细胞健康和血糖控制。

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引用次数: 0

Retraction notice to “Identification of metabolites of brexpiprazole in human urine for use in monitoring patient compliance” [Clin. Mass Spectrom. 6 (2017) 21–24] 对“鉴定人尿中brexpiprazole代谢物用于监测患者依从性”的撤回通知[临床。质谱，6 (2017)21-24 [j]

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-04-01 DOI: 10.1016/j.jmsacl.2025.03.001

Jeffrey R. Enders , Sandeep Gunna Reddy , Erin C. Strickland , Gregory L. McIntire

引用次数: 0

Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma LC-MS/MS检测尿同型香草酸和香草酸的方法及其在神经母细胞瘤诊断中的应用

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-04-01 DOI: 10.1016/j.jmsacl.2025.04.007

Lucilla Rossi , Yvette A.H. Matser , Sebastiano Barco , Alessia Cafaro , Federica Pigliasco , Margherita Biondi , Fabrizio Mancin , Maria van der Ham , Monique G.M. de Sain-van der Velden , Shifra Ash , Maja Beck Popovic , André B.P. van Kuilenburg , Massimo Conte , Alberto Garaventa , Godelieve A.M. Tytgat , Giuliana Cangemi

Background

Urinary catecholamine metabolites are well-established biomarkers for neuroblastoma (NB). Homovanillic acid (HVA) and vanillylmandelic acid (VMA) are the most frequently measured metabolites within SIOPEN – Catecholamine Working Group laboratories. Here, we evaluated the performance of a new LC-MS/MS in vitro diagnostic (IVD) kit for HVA and VMA to facilitate inter-laboratory harmonization.

Methods

HVA and VMA and their deuterated internal standards were analyzed with a commercial method, on a ThermoFisher Quantiva LC-MS/MS. Validation was performed first using internal quality control and external quality assessment (IQC and EQA) samples. Next by clinical validation on 120 samples, previously tested by HPLC-ECD. Finally, 36 samples were exchanged between SIOPEN reference laboratories and analyzed by three methods.

Results

Using QCs and EQA the method was validated in a wide calibration range (4.61–830 µmol/L for HVA and 4.44–800 µmol/L for VMA). Intra-day CVs (n = 5) were 7 and 8 % for HVA and 5 and 6 % for VMA for QC low and QC high, respectively; Inter-day CV% were 7 and 3 % for HVA and 2 and 7 % for VMA at QC low and QC high, respectively. Its application to 120 clinical samples confirmed a high diagnostic accuracy. The inter-laboratory quality control assessment showed interchangeable results (p = 0,73 and p = 0.15 for HVA and VMA, respectively).

Conclusion

The LC-MS/MS IVD method could be considered a useful tool for clinical laboratories involved in the measurement of catecholamines, contributing to harmonization efforts.

尿儿茶酚胺代谢物是神经母细胞瘤（NB）公认的生物标志物。高香草酸（HVA）和香草扁桃酸（VMA）是SIOPEN -儿茶酚胺工作组实验室中最常测量的代谢物。在这里，我们评估了一种新的LC-MS/MS体外诊断（IVD）试剂盒对HVA和VMA的性能，以促进实验室间的协调。方法采用商用方法，在ThermoFisher Quantiva LC-MS/MS上对shva和VMA及其氘化内标进行分析。首先使用内部质量控制和外部质量评估（IQC和EQA）样品进行验证。接下来对120个样品进行临床验证，之前通过HPLC-ECD检测。最后，36份样品在SIOPEN参考实验室间交换，采用三种方法进行分析。结果采用质谱仪和EQA验证了该方法的校准范围（HVA为4.61 ~ 830µmol/L， VMA为4.44 ~ 800µmol/L）。QC低和QC高的HVA和VMA的日内cv （n = 5）分别为7%和8%和5%和6%；在QC低和QC高时，HVA的日间CV分别为7%和3%，VMA的日间CV分别为2%和7%。对120个临床样本的应用证实了其较高的诊断准确性。实验室间质量控制评价结果可互换（HVA和VMA分别为p = 0,73和p = 0.15）。结论LC-MS/MS IVD方法可作为临床实验室儿茶酚胺含量测定的有效工具，有助于儿茶酚胺含量的统一。

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引用次数: 0

Quantification of total sBCMA in human plasma by peptide-level immunocapture LC-MS/MS 肽水平免疫捕获LC-MS/MS定量人血浆中总sBCMA

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-04-01 DOI: 10.1016/j.jmsacl.2025.04.006

Johanna Paris , Alexandra Tavernier , Sylvie Bethegnies, Sandrine Descloux, Olivier Fedeli

Background

B-cell maturation antigen (BCMA) is a membrane protein that is overexpressed in multiple myeloma cells and can be targeted with biotherapeutic agents. BCMA is naturally shed by γ-secretase, leading to the formation of soluble BCMA (sBCMA), which circulates in the blood. sBCMA can affect the efficacy of BCMA-targeted therapies and act as a drug sink. Additionally, sBCMA can interfere with pharmacokinetic measurements when BCMA is directly targeted. Therefore, quantification of this biomarker during clinical trials is essential to assess the effective dose and understand pharmacokinetic results. When quantifying sBCMA using ligand binding assays or hybrid assays, the biotherapeutic can interfere with the capture of sBCMA, leading to an underestimation of its levels.

Methods

Samples were denatured, reduced, and alkylated prior to trypsin digestion. sBCMA peptide enrichment was performed using anti-peptide polyclonal antibodies. Reversed-phase chromatographic separation was conducted on a biocompatible C18 column with an analysis time of sixteen minutes per sample. The SCIEX QTRAP 5500 mass spectrometer operated in multiple reaction monitoring mode. The calibration curve was prepared by spiking recombinant sBCMA into monkey plasma.

Results

The parallelism between the authentic and surrogate matrices, as well as between the endogenous and recombinant proteins, was validated. Comparisons were made between protein and peptide level hybrid assays, with the peptide level approach effectively removing the interference of the biotherapeutic. Additionally, the peptide level immunocapture LC-MS/MS demonstrated ligand tolerance.

Conclusion

The peptide level immunocapture LC-MS/MS analysis eliminated the interference of anti-BCMA biotherapeutics, allowing for the quantification of total sBCMA in clinical samples while achieving a LLOQ of 10 ng/mL.

b -细胞成熟抗原（BCMA）是一种在多发性骨髓瘤细胞中过表达的膜蛋白，可作为生物治疗药物的靶点。BCMA通过γ-分泌酶自然脱落，形成可溶性BCMA (sBCMA)，在血液中循环。sBCMA会影响bcma靶向治疗的疗效，起到药物沉淀的作用。此外，当BCMA直接靶向时，sBCMA会干扰药代动力学测量。因此，在临床试验中对该生物标志物进行量化对于评估有效剂量和了解药代动力学结果至关重要。当使用配体结合测定法或杂交测定法定量sBCMA时，生物疗法会干扰sBCMA的捕获，导致对其水平的低估。方法在胰蛋白酶消化前对样品进行变性、还原和烷基化处理。利用抗多肽多克隆抗体对sBCMA肽进行富集。在生物相容性C18色谱柱上进行反相色谱分离，每个样品的分析时间为16分钟。SCIEX QTRAP 5500质谱仪在多反应监测模式下工作。将重组sBCMA注入猴子血浆制备校准曲线。结果验证了原基质与替代基质、内源蛋白与重组蛋白的相似性。比较蛋白质水平和多肽水平的杂交测定，多肽水平的方法有效地消除了生物治疗的干扰。此外，肽水平免疫捕获LC-MS/MS显示配体耐受性。结论多肽水平免疫捕获LC-MS/MS分析消除了抗bcma生物治疗药物的干扰，可以定量测定临床样品中总sBCMA， LLOQ为10 ng/mL。

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引用次数: 0

A rapid method for determination of rosuvastatin in blood plasma with supported liquid extraction 支撑液体萃取法快速测定血浆中瑞舒伐他汀的含量

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-04-01 DOI: 10.1016/j.jmsacl.2025.04.003

Tjaša Dermota , Mojca Božič Mijovski , Jurij Trontelj

Introduction

Accurate measurement of rosuvastatin in plasma is critical for effective patient management and treatment monitoring following myocardial infarction (MI). Expensive solid-phase extraction (SPE) and time-consuming liquid–liquid extraction (LLE) have been established for quantifying rosuvastatin. Supported liquid extraction (SLE) could offer a rapid, cost-effective alternative.

Objectives

This study aimed to develop and validate a rapid, cost-effective, accurate, and precise method for quantifying rosuvastatin in high-dose plasma samples from patients following MI.

Methods

Rosuvastatin was extracted from EDTA plasma using SLE and quantified with LC-MS/MS with positive electrospray ionization. The method was validated according to ICH M10 guidelines, focusing on selectivity, matrix effect, accuracy, precision, linearity, and carryover. Rosuvastatin-D6 was used as an internal standard. Additionally, thirty plasma samples from patients on high-dose rosuvastatin therapy (20 or 40 mg/day) following MI were analyzed by both LLE and SLE methods and compared.

Results

The method was successfully validated, demonstrating linearity across a range of 0.1 ng/mL to 50 ng/mL. Compared to the LLE method, SLE achieved superior extraction recovery (96.3 % vs. 60 %) and precision (RSD: 11.9 % vs. 13.6 %) at 0.3 ng/mL rosuvastatin, with a lower absolute matrix effect (12.7 % vs. −36.7 %). Accuracy was comparable (109.3 % vs. 92.8 %). Although SLE involves higher initial costs, it significantly enhances throughput, reduces solvent usage, and minimizes contamination and equipment wear.

Conclusion

This study validates SLE as a superior method for quantifying rosuvastatin in plasma, outperforming LLE in recovery, reproducibility, and automation. SLE offers greater accuracy and reliability, making it ideal for high-throughput applications.

准确测量血浆瑞舒伐他汀对心肌梗死（MI）后患者的有效管理和治疗监测至关重要。建立了昂贵的固相萃取法（SPE）和耗时的液液萃取法（LLE）来定量瑞舒伐他汀。支持液体萃取（SLE）是一种快速、经济的替代方法。目的建立并验证一种快速、经济、准确、精确的方法，用于急性心肌梗死患者高剂量血浆样品中瑞舒伐他汀的定量。方法采用SLE法从EDTA血浆中提取瑞舒伐他汀，并采用电喷雾正离子化LC-MS/MS定量。根据ICH M10指南对该方法进行验证，重点关注选择性、基质效应、准确度、精密度、线性和结转。以瑞舒伐他汀d6为内标。此外，采用LLE和SLE方法对心肌梗死后接受高剂量瑞舒伐他汀治疗（20或40 mg/天）的患者的30份血浆样本进行分析和比较。结果该方法在0.1 ng/mL ~ 50 ng/mL范围内呈线性关系。与LLE法相比，0.3 ng/mL瑞舒伐他汀的SLE提取回收率（96.3% vs. 60%）和精密度（RSD: 11.9% vs. 13.6%）更高，绝对基质效应更低（12.7% vs.−36.7%）。准确率相当（109.3%对92.8%）。虽然SLE涉及较高的初始成本，但它显著提高了产量，减少了溶剂的使用，并最大限度地减少了污染和设备磨损。结论本研究验证了SLE作为血浆中瑞舒伐他汀定量的优越方法，在回收率、重现性和自动化方面优于LLE。SLE提供更高的准确性和可靠性，使其成为高通量应用的理想选择。

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引用次数: 0

Development of an isotope dilution gas chromatography − mass spectrometry candidate reference measurement procedure for glucose in human serum 建立一种同位素稀释气相色谱-质谱法测定人血清中葡萄糖的候选参考方法

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-04-01 DOI: 10.1016/j.jmsacl.2025.04.005

Komal Dahya , Heather C. Kuiper , Sarah W. Kingsley , Uliana Danilenko , Hubert W. Vesper

Introduction

Diabetes is the seventh leading cause of death in the United States, impacting over 37 million people. Accurate glucose measurements are critical for effective diabetes management. A reliable candidate reference measurement procedure (cRMP) for assessing the analytical performance of glucose tests performed in patient care is essential for ensuring measurement accuracy.

Methods

We have developed a gas chromatography-mass spectrometry (GC–MS)-based cRMP for glucose in human serum. In this procedure, glucose is measured as the aldononitrile acetate derivative and quantitated using a ¹³C₆-glucose internal standard.

Results

Analytical selectivity was achieved through chromatographic separation and monitoring the quantitation ion/confirmation ion ratios in samples. With bias ranging from −0.79 % to 0.67 % for eight levels of serum-based certified reference materials from the National Institute of Standards and Technology (NIST) and Laboratoire national de métrologie et d’essais (LNE) and total CVs of 1.11 %, 0.68 % and 0.74 % at the low, medium, and high glucose concentration levels, respectively, the cRMP provided excellent accuracy and precision. The calibration curve was linear throughout the 13.51–378.21 mg/dL [0.75–21 mmol/L] measurement range (R² = 0.9999), with a mean slope of 270.73 (95 % CI, 270.19 to 271.27) and an intercept of 0.021 (95 % CI, −0.157 to 0.199). The limit of detection was 0.25 mg/dL (0.014 mmol/L) and the limit of quantitation was 0.83 mg/dL (0.046 mmol/L).

Conclusion

The described GC–MS method, with metrological traceability to the International System of Units (SI), provides highly accurate and precise measurements of glucose in human serum.

糖尿病是美国第七大死因，影响了超过3700万人。准确的血糖测量对有效的糖尿病管理至关重要。一种可靠的候选参考测量程序（cRMP）用于评估在患者护理中进行的葡萄糖测试的分析性能，对于确保测量准确性至关重要。方法建立了以气相色谱-质谱（GC-MS）为基础的血清葡萄糖定量分析方法。在这个过程中，葡萄糖被测量为乙腈乙酸酯衍生物，并使用13c6 -葡萄糖内标进行定量。结果通过色谱分离和监测样品的定量/确认离子比，实现了分析选择性。来自美国国家标准与技术研究所（NIST）和美国国家实验室（LNE）的8个水平的血清认证标准物质的偏倚范围为- 0.79%至0.67%，在低、中、高葡萄糖浓度水平下的总cv分别为1.11%、0.68%和0.74%，cRMP提供了极好的准确度和精密度。在13.51 ~ 378.21 mg/dL [0.75 ~ 21 mmol/L]测量范围内，校准曲线呈线性关系（R2 = 0.9999），平均斜率为270.73 (95% CI, 270.19 ~ 271.27)，截距为0.021 （95% CI, - 0.157 ~ 0.199）。检测限为0.25 mg/dL (0.014 mmol/L)，定量限为0.83 mg/dL （0.046 mmol/L）。结论所建立的气相色谱-质谱联用方法可溯源至国际单位制（SI），对人血清中葡萄糖的测定具有较高的准确度和精密度。

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引用次数: 0

Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocin 一种高灵敏度、定量LC-MS/MS检测血浆催产素的方法的开发和验证

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-02-22 DOI: 10.1016/j.jmsacl.2025.02.002

E. Grifnée , A. Mackowiak , J. Demeuse , M. Schoumacher , L. Huyghebaert , W. Determe , T. Dubrowski , P. Massonnet , S. Peeters , G. Scantamburlo , E. Cavalier , C.Le Goff

Introduction

Oxytocin is a 9-amino acid peptide that serves as neuromodulator in the human central nervous system. This peptide is implicated in the regulation of diverse behaviors and plays a significant role in positive social interaction. Currently, oxytocin levels are measured using immunoassays. However, these methods have several limitations that can lead to false results and erroneous interpretation. Given the remarkably low endogenous level of oxytocin in human plasma (low ng/L levels), we developed and rigorously validated a novel and highly sensitive LC-MS/MS method for oxytocin quantification in plasma.

Methods

Oxytocin was initially extracted using solid-phase extraction with an Oasis HLB 30 mg plate and then subjected to LC-MS/MS analysis. PBS-0.1 % BSA served as surrogate matrix for the preparation of validation samples and the calibration curve, ensuring no endogenous interference. The validation design followed the Clinical Laboratory Standards Institute guidelines. Precision, accuracy, and measurement uncertainty were determined using single-nested analysis of variance and e.noval software.

Results

A lower limit of quantification of 1 ng/L was achieved. The method was validated for oxytocin concentrations ranging from 1 ng/L to 75 ng/L, with precision (coefficient of variation) below 10 %, accuracy ranging from 94 % to 108 %, and measurement uncertainty below 15 %.

Conclusion

In this work, we developed and validated a highly sensitive LC-MS/MS method for the quantification of oxytocin in plasma. Our novel methodology is well-suited for clinical applications.

催产素是一种9个氨基酸的肽，在人类中枢神经系统中起神经调节剂的作用。这种肽与多种行为的调节有关，在积极的社会互动中起着重要作用。目前，催产素水平是通过免疫分析法来测量的。然而，这些方法有一些局限性，可能导致错误的结果和错误的解释。鉴于人血浆中催产素的内源性水平非常低（低ng/L水平），我们开发并严格验证了一种新的、高灵敏度的LC-MS/MS定量血浆中催产素的方法。方法采用Oasis HLB 30 mg平板固相萃取法提取催产素，然后进行LC-MS/MS分析。以pbs - 0.1%牛血清白蛋白为替代基质制备验证样品和校准曲线，确保无内源干扰。验证设计遵循临床实验室标准协会的指南。精密度、准确度和测量不确定度采用单套方差分析和e.noval软件确定。结果达到了1 ng/L的定量下限。该方法在1 ~ 75 ng/L的浓度范围内验证，精密度（变异系数）在10%以下，准确度在94% ~ 108%之间，测量不确定度在15%以下。结论建立并验证了血浆中催产素的高灵敏度LC-MS/MS定量方法。我们的新方法非常适合临床应用。

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引用次数: 0

Barcoded labels as potential source of zinc contamination in trace metal testing 条形码标签是痕量金属检测中锌污染的潜在来源

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-02-21 DOI: 10.1016/j.jmsacl.2025.02.003

Difei Sun , Donna Sealy , Dorothy Truong , Danijela Konforte

引用次数: 0

MRM-based LC-MS method for accurate C-peptide quantitation 基于mrm的LC-MS方法用于c肽的准确定量

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Mass Spectrometry and Advances in the Clinical Lab

Pub Date : 2025-02-12 DOI: 10.1016/j.jmsacl.2025.02.001

Will Grothoff, Ivan Khodakivskyi, Aleks Shin, Randie Little, Shawn Connolly, Kuanysh Kabytaev

Introduction

C-peptide secretion mirrors beta-cell function and has emerged as a valuable clinical biomarker for diabetes mellitus. C-peptide measurements can provide estimates of insulin secretory capacity, aiding in clinical decision-making and differentiation between diabetes types. Unfortunately, C-peptide assays are still not standardized, which may limit their practical clinical use. We have developed an MRM-based LC-MS method that demonstrated accuracy close to our reference method.

Objective

To develop and validate a mass spectrometry method for accurate quantitation of C-peptide.

Method

A serum sample was spiked with isotope-labeled C-peptide as a standard. The enrichment process involved protein precipitation with methanol, solid-phase extraction, and anion exchange for C-peptide enrichment followed by Glu-C digestion. The peptide LGGGPGAGSLQPLALE was quantitated using MRM in positive ion mode. The calibration process includes C-peptide CRM material to ensure a complete traceability chain for the measurement.

Results

The assay exhibited linearity across a wide range of C-peptide concentrations and a limit of quantitation of 0.058 nmol/L. The inter-day imprecision was less than 9.6 % CV, and the intra-day imprecision was less than 8.9 % CV. Spiking with bilirubin, triglycerides, and hemoglobin demonstrated no interference, except for triglycerides at very high levels. The method exhibited a strong correlation to the C-peptide reference method (r2 = 0.95).

Conclusion

The developed mass spectrometry method has demonstrated accurate results in C-peptide quantitation and can serve as a supplemental method to the existing C-peptide reference method. This ensures sustained stability over time and ultimately refines the existing reference system.

c肽分泌反映了β细胞的功能，已成为糖尿病的一种有价值的临床生物标志物。c肽测量可以提供胰岛素分泌能力的估计，帮助临床决策和区分糖尿病类型。不幸的是，c肽检测仍然没有标准化，这可能限制了它们的实际临床应用。我们开发了一种基于mrm的LC-MS方法，其精度接近我们的参考方法。目的建立并验证c肽的质谱测定方法。方法用同位素标记的c肽加标法测定血清样品。富集过程包括甲醇沉淀蛋白质、固相萃取、阴离子交换富集c肽，然后进行Glu-C消化。LGGGPGAGSLQPLALE肽在正离子模式下用MRM定量。校准过程包括c肽CRM材料，确保测量有完整的可追溯链。结果该方法在较宽的c肽浓度范围内呈线性，定量限为0.058 nmol/L。日间不精密度小于9.6% CV，日间不精密度小于8.9% CV。与胆红素、甘油三酯和血红蛋白一起注射没有干扰，除非甘油三酯水平很高。该方法与c肽参考方法具有较强的相关性（r2 = 0.95）。结论建立的质谱分析方法对c肽的定量结果准确，可作为现有c肽参比法的补充。这确保了随着时间的推移持续稳定，并最终完善了现有的参考系统。

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引用次数: 0