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Method for assaying methylmalonic acid, methylcitric acid and/or homocysteine by liquid chromatography tandem mass spectrometry

A technology for homocysteine ​​and methylmalonic acid, applied in the field of liquid chromatography tandem mass spectrometry, can solve the problems of simultaneous detection of methylmalonic acid, long detection time, large amount of detection samples, etc., and achieve operational And the effect of clear response, short detection time and saving sample volume

Inactive Publication Date: 2017-08-25
浙江博圣生物技术股份有限公司
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Problems solved by technology

[0008] The technical problem to be solved by the present invention is that in the prior art, methylmalonic acid, methylcitric acid and homocysteine ​​cannot be detected at the same time, and the amount of detection samples is large and the detection time is long. Method for the detection of methylmalonate, methylcitrate and / or homocysteine ​​in urine, plasma or blood spots by mass spectrometry

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  • Method for assaying methylmalonic acid, methylcitric acid and/or homocysteine by liquid chromatography tandem mass spectrometry
  • Method for assaying methylmalonic acid, methylcitric acid and/or homocysteine by liquid chromatography tandem mass spectrometry

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Embodiment 1

[0082]Example 1 Method for detecting methylmalonic acid, methylcitrate and homocysteine ​​in blood spots by liquid chromatography tandem mass spectrometry

[0083] see figure 1 , Example 1 provides a specific implementation process for detecting methylmalonic acid, methylcitric acid and homocysteine ​​in blood spots by liquid chromatography tandem mass spectrometry.

[0084] (1) Pretreatment-derivatization reaction

[0085] Select one well of a 96-well plate, and punch four small blood spots with a diameter of 3 mm. Add 150ul extractant solution (DTT solution containing 3d-methylmalonic acid, 3d-methylcitric acid and 8d-homocysteine), incubate and shake for 0.5 hour, let stand for 1min. Transfer the liquid to another clean new 96-well plate, and dry it with nitrogen at 40°C. Add 100 μl of n-butanol hydrochloride, shake at a constant temperature of 65° C. and 600 rpm / min, and react for 15 minutes. Let stand for 3mins, blow dry with nitrogen at 40°C. Add 100 μl of 30% acet...

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Abstract

The invention discloses a method for assaying methylmalonic acid, methylcitric acid and / or homocysteine by liquid chromatography tandem mass spectrometry. The assay method includes sample pretreatment (derivation reaction) and assay of to-be-assayed sample by liquid chromatography tandem quadrupole mass spectrometry, a cation mode is adopted, and multiple-reaction monitoring (MRM) is adopted as a scanning mode. The method provided by the invention can simultaneously or respectively assay three types of substances with great polarity difference, i.e. methylmalonic acid, methylcitric acid and homocysteine, in a sample, the amount of the used sample is small, the assay time is short, the flux is high, the assay precision is high, substance coverage is wide, and the cost is low.

Description

technical field [0001] The invention relates to the technical field of liquid chromatography tandem mass spectrometry detection, in particular to a method for detecting methylmalonic acid, methylcitric acid and / or homocysteine ​​in urine, plasma or blood spots by liquid chromatography tandem mass spectrometry. Background technique [0002] Methylmalonate is a bypass product of methylmalonyl-CoA under abnormal metabolism in the decomposition pathway of valine, methionine, threonine, isoleucine, cholesterol and odd-numbered chain fatty acids. The metabolism of methylmalonyl-CoA caused by the decreased activity of methylmalonyl mutase or methylcobalamin caused by the mutation is blocked, resulting in the abnormal accumulation of metabolites such as methylmalonate and methylcitrate, causing Brain, liver, kidney and other multiple organ damage, the above conditions are clinically classified as methylmalonic acidemia. [0003] Methylmalonic acidemia is an autosomal recessive gene...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/7266
Inventor 王军娟赵珺吴敏航黄成刚俞晓敏
Owner 浙江博圣生物技术股份有限公司
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