Development and validation of a LC-MS/MS method for quantitation of 3-hydroxypentanoic acid and 3-oxopentanoic acid in human plasma and its application to a clinical study of glucose transporter type I deficiency (G1D) syndrome.
Development and validation of a LC-MS/MS method for quantitation of 3-hydroxypentanoic acid and 3-oxopentanoic acid in human plasma and its application to a clinical study of glucose transporter type I deficiency (G1D) syndrome.
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DOI：
10.1016/j.jpba.2021.114335
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发表时间：
2021-10-25
影响因子：
3.4
通讯作者：
Putnam WC
中科院分区：
文献类型：
--
作者：
Kallem RR;Primeaux S;Avila A;Pascual JM;Putnam WC
在人类和实验动物代谢的底物含有奇数个碳能够燃料的三羧酸循环，如庚酸的兴趣，促使我们开发和验证一种选择性和特定的液相色谱串联质谱方法，同时，定量测定血浆中的酮体副产物3-羟基戊酸和3-氧代戊酸。人血浆样品用含0.2%甲酸的甲醇进行蛋白沉淀。在Phenomenex Luna C18色谱柱上，采用含0.1%甲酸的水和含0.1%甲酸的甲醇的移动的流动相以0.3 mL/min流速进行梯度洗脱，实现色谱分离度。3-羟基戊酸、3-氧代戊酸和舒巴坦（内标）的保留时间分别为3.85、4.23和5.11 min。根据美国食品药品监督管理局指南进行验证。3-羟基戊酸的验证范围为0.078-5 μg/mL，3-氧代戊酸的验证范围为0.156-10 μg/mL。该方法在该范围内具有准确度和精密度，并在人血浆中显示出10倍稀释度。分析物和内标物的回收率>88%。在人血浆中未观察到基质效应。3-羟基戊酸和3-氧代戊酸在包括自动进样器、台式和冻融在内的条件下均稳定，并在-80 °C下表现出长期稳定性。该方法应用于测量接受甘油三酯三庚酸甘油酯（作为庚酸的来源）的受试者的血浆中3-羟基戊酸和3-氧代戊酸浓度，用于葡萄糖转运蛋白I型缺乏症（G1 D）综合征的实验性治疗。
Interest in the human and experimental animal metabolism of substrates containing an odd number of carbons capable of fueling the tricarboxylic acid cycle such as heptanoic acid has motivated us to develop and validate a selective and specific liquid chromatography tandem mass spectrometric method for the simultaneous, quantitative determination of the ketone body byproducts 3-hydroxypentanoic acid and 3-oxopentanoic acid in plasma. Human plasma samples were protein-precipitated with methanol containing 0.2% formic acid. Chromatographic resolution was achieved on a Phenomenex Luna C18 column using gradient elution with mobile phases of water containing 0.1% formic acid and methanol containing 0.1% formic acid at 0.3 mL/min flow rate. The retention times of 3-hydroxypentanoic acid, 3-oxopentanoic acid and sulbactam (internal standard) were 3.85, 4.23, and 5.11 min, respectively. Validation was conducted in accordance with United States Food and Drug Administration guidance. The validated range of 3-hydroxypentanoic acid was 0.078–5 μg/mL and 0.156–10 μg/mL for 3-oxopentanoic acid. The method was accurate and precise over this range and exhibited 10-fold dilution integrity in human plasma. Recovery >88% was achieved for analytes and internal standard. There was no matrix effect observed in human plasma. Both 3-hydroxypentanoic acid and 3-oxopentanoic acid were stable across conditions including autosampler, benchtop and freeze-thaw, as well as demonstrated long-term stability at –80 °C. The method was applied to the measurement of 3-hydroxypentanoic acid and 3-oxopentanoic acid concentrations in plasma from subjects receiving the triglyceride triheptanoin (as a source of heptanoate) for the experimental treatment of glucose transporter type I deficiency (G1D) syndrome.
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DOI：
10.1152/ajpendo.00384.2009
发表时间：
2010-02-01
影响因子：
5.1
作者：
Gu, Lei;Zhang, Guo-Fang;Brunengraber, Henri
通讯作者：
Brunengraber, Henri
DOI：
10.1152/ajpendo.00366.2005
发表时间：
2006-10-01
影响因子：
5.1
作者：
Kinman, Renee P.;Kasumov, Takhar;Brunengraber, Henri
通讯作者：
Brunengraber, Henri
DOI：
10.1152/ajpendo.1995.268.3.e446
发表时间：
1995-03-01
影响因子：
5.1
作者：
LECLERC, J;ROSIERS, CD;BRUNENGRABER, H
通讯作者：
BRUNENGRABER, H
影响因子：
1.8
作者：
Jaochico, Allan;Sangaraju, Dewakar;Shahidi-Latham, Sheerin K.
通讯作者：
Shahidi-Latham, Sheerin K.
影响因子：
29
作者：
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通讯作者：
Lu, Hanzhang