Detection kit based on tandem mass spectrum non-derivatization method

A detection kit and tandem mass spectrometry technology, which is applied in the field of tandem mass spectrometry derivatization detection kits, to achieve the effects of reducing social burden, improving population quality, and simple operation

Active Publication Date: 2015-05-27
上海敬元投资(集团)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a tandem mass spectrometry derivatization detection kit to solve the defects in the existing LC-MS / MS detection

Method used

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  • Detection kit based on tandem mass spectrum non-derivatization method
  • Detection kit based on tandem mass spectrum non-derivatization method
  • Detection kit based on tandem mass spectrum non-derivatization method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Embodiment 1, determination of extract formula

[0066] Take the same quality control blood sheet, respectively use the extracts of the formulations described in Table 2, and detect the content of amino acids, carnitine and acylcarnitine according to the above-mentioned tandem mass spectrometry derivatization method, the mobile phase is methanol: water: formic acid volume ratio is 1000 :10:0.5.

[0067] Table 2 (volume ratio)

[0068] Extract

[0069] Results: The extraction solution is methanol: water: formic acid with a volume ratio of 1000:1~50:0.01~2, which can achieve ideal results (Table 3, 4), proper hemolysis and protein precipitation (Figure 8), and the recovery rate and accuracy rate can be achieved. It meets the quality control requirements of the US CDC. If it exceeds the range of the formula, such as formula 5 and 6, the hemolysis is too light or too heavy, resulting in a decrease or increase in the recovery rate of some amino acids or acylcarni...

Embodiment 2

[0076] Embodiment 2, determination of mobile phase formulation

[0077] Take the same quality control blood piece, and detect the amino acid content according to the derivatization method of the above-mentioned tandem mass spectrometry, except that the mobile phase is different, the mobile phase of the formula described in Table 5 is used respectively, and the extraction solution is methanol: water: formic acid volume ratio is 1000:10: 1. At the same time, use the non-derivatization method kit from PerkinElmer (NeoBase) to detect the same quality control blood film, and obtain the 95% confidence limit of each target component (PE kit 95% confidence limit)

[0078] Table 5 (volume ratio)

[0079]

[0080]

[0081] Results: The ratio of acid to water in the mobile phase affects the ionization of the analytes. The mobile phase is methanol: water: formic acid (1000:1~50:0.01~2) to obtain ideal detection results, and the relative balance of recovery rates of various amino a...

Embodiment 3

[0085] Example 3. A detection kit for tandem mass spectrometry derivatization method

[0086] 3.1 A tandem mass spectrometry derivatization detection kit comprising:

[0087] 1 bottle of isotope-labeled amino acid reference substance mixture;

[0088] 1 bottle of isotope-labeled acetylcarnitine reference substance mixture;

[0089] Extract (volume ratio of methanol: water: formic acid is 1000:1:0.1) 1 bottle (150ml);

[0090] Mobile phase (methanol: water: formic acid volume ratio is 1000:1:0.1) 1 bottle (400ml);

[0091] 10 U-bottom heat-resistant 96-well extraction plates;

[0092] V-bottom 96-well detection plate 10 pieces;

[0093] Sticky extraction plate film 20 pieces;

[0094] 20 pieces of aluminum foil for detection board.

[0095] 3.2 A tandem mass spectrometry derivatization detection kit comprising:

[0096] 1 bottle of isotope-labeled amino acid reference substance mixture;

[0097] 1 bottle of isotope-labeled acetylcarnitine reference substance mixture;

...

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Abstract

The invention relates to the technical field of chemical analysis and detection, and in particular relates to a detection kit based on a tandem mass spectrum non-derivatization method. The detection kit comprises an isotope labeled amino acid standard substance mixture, an isotope labeled acyl carnitine standard substance mixture, an extract liquor and a flow phase. The detection kit based on the tandem mass spectrum non-derivatization method is characterized in that the extract liquor consists of methyl alcohol, water and saturated monocarboxylic acid, wherein the volume ratio of the methyl alcohol to the water to the saturated monocarboxylic acid is 1000: (1-50): (0.01-2). The detection kit based on the tandem mass spectrum non-derivatization method provided by the invention is high in flexibility and accuracy rate, is simple to operate, and has important significance for improving the diagnosis level of metabolic diseases, lowering the burden of society and families and improving the quality of population.

Description

technical field [0001] The invention relates to the technical field of chemical analysis and detection, in particular to a tandem mass spectrometry derivatization detection kit. Background technique [0002] A variety of amino acids and acylcarnitines in blood are the marker products of amino acid, organic acid and fatty acid metabolism in the human body. Changes in amino acid spectrum and acylcarnitine spectrum can occur in the blood of metabolic diseases. Liquid phase tandem mass spectrometry (LC- MS / MS) has been applied in the screening of newborn genetic metabolic diseases and the diagnosis of clinical metabolic diseases. [0003] Tandem mass spectrometry (MS / MS) means that two mass spectrometers are connected in series to perform secondary mass spectrometry detection. Using the tandem mass spectrometry technology of ultrasensitivity, high specificity, high selectivity and rapid detection, it can detect a The samples are analyzed for dozens of metabolites. Through the a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/04G01N30/02
Inventor 龚振华
Owner 上海敬元投资(集团)有限公司
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