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

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。

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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

Rapid identification of SARS CoV-2 omicron sub-variant JN.1 (BA.2.86.1.1) with mass spectrometry 利用质谱法快速鉴定 SARS CoV-2 omicron 亚变体 JN.1 (BA.2.86.1.1)

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.2024.08.003

Henry E. Lanyon, Kevin M. Downard

Objective

The rapid detection and differentiation of strains of the BA.2.86 lineage including the new sub-variant JN.1 (BA.2.86.1.1) is demonstrated employing selected ion monitoring (SIM) and high resolution mass spectrometry.

Methods

A study of a preliminary set of BA.2.86 lineage positive specimens, identified BA.2.86 and BA.2.86.1.1 peptide markers in 62.5 % and 29.1 % of samples.

Results

Peptide-specific markers in the surface spike protein associated with the L455S mutation are confidently detected with high sensitivity in protein and virus digests.

The virus was thus confidently assigned in over 91 % of positive specimens.

Conclusions

A rise in the global prevalence of the JN.1 (BA.2.86.1.1) immune evasive sub-variant, that emerged in late 2023, requires that new strategies and protocols to detect such strains in human specimens are accelerated and implemented.

目的利用选择离子监测（SIM）和高分辨质谱法证明了对 BA.2.86 系菌株（包括新的亚变异株 JN.1（BA.2.86.1.1））的快速检测和区分。结果与 L455S 突变相关的表面尖峰蛋白中的肽特异性标记在蛋白质和病毒消化液中被高灵敏度地检测到。(结论 2023 年末，JN.1（BA.2.86.1.1）免疫逃避亚变异株在全球的流行率上升，这要求加快并实施在人类样本中检测此类毒株的新策略和方案。

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引用次数: 0

Isotope-dilution-LC-MS/MS candidate reference measurement procedure for cefepime in human serum 人血清中头孢吡肟的同位素稀释-LC-MS/MS 候选参考测量程序

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.2024.08.001

Judith Schäffler, Michael Vogeser, Katharina Habler

Background

Reference measurement procedures are an essential element in the standardization and comparability of analytical measurement results in laboratory medicine. No LC-MS/MS-based reference measurement procedure for cefepime in serum has been published previously.

Materials and methods

An isotope-dilution based two-dimensional LC-MS/MS reference measurement procedure for cefepime concentrations in human serum was developed and tested. The value assignment of unknown samples is based on a defined measurement series validation. Six unknown samples can be measured per series. Pass criteria for the run and the samples were determined empirically based on a performance evaluation. For this purpose, a between-run determination of five runs of the defined measurement series with six cefepime samples was carried out and evaluated. The goal was to define rigorous, realistic target limits and minimize measurement uncertainty. The final defined target limits are used for series-based validation and value assignment. The results for the six unknown samples are provided with the associated measurement uncertainty for this series.

Results

The developed and extensively studied measurement procedure for the quantification of cefepime in serum was found to be practicable and fit for its purpose. The between-run mean imprecision of the six cefepime samples was ≤ 2.0 %, for the QCs it was ≤ 2.3 % and the between-run mean inaccuracy of the QCs was within ± 1.1 %.

Conclusion

The novel isotope-dilution-LC-MS/MS measurement procedure in accordance to ISO 15193 can be recommended as candidate reference measurement procedure for the value assignment of cefepime concentrations in human serum.

背景参考测量程序是实验室医学分析测量结果标准化和可比性的基本要素。材料与方法开发并测试了一种基于同位素稀释的二维 LC-MS/MS 参考测量程序，用于测量人血清中头孢吡肟的浓度。未知样品的定值基于确定的测量系列验证。每个系列可测量六个未知样品。运行和样品的通过标准是根据性能评估经验确定的。为此，对包含六个头孢吡肟样品的已定义测量系列的五次运行进行了运行间测定和评估。目的是确定严格、实际的目标限值，并将测量的不确定性降至最低。最终确定的目标限值用于基于系列的验证和赋值。结果经广泛研究发现，所开发的血清中头孢吡肟定量测量程序切实可行，符合其目的。六份头孢吡肟样品的运行间平均不精确度≤ 2.0%，质控品的运行间平均不精确度≤ 2.3%，质控品的运行间平均不准确度在 ± 1.1% 以内。结论符合 ISO 15193 标准的新型同位素稀释-LC-MS/MS 测量程序可推荐用作人血清中头孢吡肟浓度定值的候选参考测量程序。

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引用次数: 0

Impact of internal standard selection on measurement results for long chain fatty acids in blood 内标选择对血液中长链脂肪酸测量结果的影响

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.2024.07.002

John M. Goodwin VII, Heather C. Kuiper, Barrett Brister, Hubert W. Vesper

Introduction

Internal standards correct for measurement variation due to sample loss. Isotope labeled analytes are ideal internal standards for the measurement of fatty acids in human plasma but are not always readily available. For this reason, quantification of multiple analytes at once is most often done using only a single or few internal standards. The magnitude of the impact this has on method accuracy and precision is not well studied for gas chromatography-mass spectrometry systems.

Objective

This study aims to estimate bias and changes in uncertainty associated with using alternative fatty acid isotopologue internal standards for the estimation of similar or dissimilar long chain fatty acids.

Method

Using a previously reported method for the quantification of 27 fatty acids in human plasma using 18 internal standards we obtained estimates of bias and uncertainty at up to three levels of fatty acid concentration.

Results

With some notable exceptions, method accuracy remained relatively stable when using an alternative internal standard (Median Relative Absolute Percent Bias: 1.76%, Median Spike-Recovery Absolute Percent Bias: 8.82%), with larger changes in method precision (Median Increase in Variance: 141%). Additionally, the degree of difference between analyte and internal standard structure was related to the magnitude of bias and uncertainty of the measurement.

Conclusion

The data presented here show that the choice of internal standard used to estimate fatty acid concentration can affect the accuracy and reliability of measurement results and, therefore, needs to be assessed carefully when developing analytical methods for the measurement of fatty acid profiles.

Disclaimer: The findings and conclusions in this report are those of the author(s) and do not necessarily represent the official position of the Centers for Disease Control and Prevention/the Agency for Toxic Substances and Disease Registry. Use of trade names is for identification only and does not imply endorsement by the Centers for Disease Control and Prevention, the Public Health Service, and the US Department of Health and Human Services.

内标可纠正因样品流失而造成的测量差异。同位素标记的分析物是测量人体血浆中脂肪酸的理想内标，但并非总能轻易获得。因此，通常只使用一种或几种内标来同时对多种分析物进行定量。对于气相色谱-质谱联用系统来说，这对方法准确度和精密度的影响程度还没有得到很好的研究。本研究旨在估算使用替代脂肪酸同位素内标估算相似或不同长链脂肪酸时的偏差和不确定性变化。我们使用以前报道过的一种方法，用 18 种内标物对人体血浆中的 27 种脂肪酸进行定量，得出了多达三个脂肪酸浓度水平下的偏差和不确定性估计值。除了一些明显的例外，使用替代内标时，方法的准确度保持相对稳定（相对绝对百分比偏差中值：1.76%，加标回收绝对百分比偏差中值：8.82%），而方法的精确度变化较大（方差增加中值：141%）。此外，分析物和内标结构的差异程度与测量的偏差和不确定性大小有关。本文提供的数据表明，用于估算脂肪酸浓度的内标物的选择会影响测量结果的准确性和可靠性，因此，在开发测量脂肪酸概况的分析方法时需要仔细评估。

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引用次数: 0