Quantitative analysis method for phosphatidylcholine in serum through capillary electrophoresis-mass spectrum and application
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A phosphatidylcholine and capillary electrophoresis technology, which is applied in the field of quantitative analysis to achieve the effects of high recovery rate and detection sensitivity, good accuracy and high accuracy
Active Publication Date: 2018-05-11
WUHAN INST OF PHYSICS & MATHEMATICS CHINESE ACADEMY OF SCI
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[0007] However, there is no report on the use of CE-MS to analyze phospholipids in serum to determine its association with the onset of Alzheimer's disease
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Embodiment 1
[0049] Based on the previous research results and a large number of experiments, this embodiment proposes a method for quantitative analysis of phosphatidylcholine in serum by capillary electrophoresis coupled with mass spectrometry (CE-MS), including the following steps:
[0050] S1. Treatment and optimization of serum samples:
[0052] Take 2μL of serum samples from the serum samples of 1114F (control group, female) and 1098F (experimental group, female) respectively, add 50ng PC standard (17:0 / 14:1, MW 717.996) as internal standard, and use PBS (1×) was diluted to 500 μL, mixed with 3 mL of chloroform / methanol=1 / 2 (v / v), vortexed and incubated on ice for 10 minutes. Add 1mL of chloroform and 1.3mL of ultrapure water, vortex at low temperature and centrifuge at 1750g, 10°C for 10 minutes, it can be seen that the interface between the two phases is clear, transfer the lower phase to a new glass tube, blow dry with n...
Embodiment 2
[0105] The application of a capillary electrophoresis-mass spectrometry (CE-MS) method for quantitative analysis of phosphatidylcholine in serum in the detection of Alzheimer's disease, the process is: (in the quantitative study of PC and LPC, we also analyzed phospholipid species at the level of individual differences).
[0106] First use the precursor ion scanning mode (Pre@184.1) to find the characteristic paired precursor ion of m / z 184.1 in the serum samples of the control group and the experimental group, and then use these 15 pairs of characteristic paired parent ion-product ion pairs within 12 weeks Four groups of serum samples collected at different times were subjected to multi-stage reaction monitoring (MRM) relative quantification. Precursor ion scan results such as Figure 10 As shown, combined with the LIPID MAPS database, except for the same structure, 13 PCs and LPCs contained in serum can be deduced (Table 8).
[0107] Table 8 Precursor ion scan mode identif...
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Abstract
The invention discloses a quantitative analysis method for phosphatidylcholine in serum through capillary electrophoresis-mass spectrum and application. The quantitative analysis method includes the steps of S1, serum sample treating, wherein a serum sample is taken and added into a phosphatidylcholine internal standard substance, methyl alcohol is directly added, incubation on ice is carried out,centrifugation is carried out, supernatant is transferred, the product is dried with nitrogen and stored, and is dissolved with the methyl alcohol before CE-MS quantitative analysis, supernatant is removed through centrifugation, and capillary electrophoresis sample feeding is prepared, wherein the phosphatidylcholine internal standard substance is phosphatidylcholine 17:0 / 14:1, MW 717.996; S2, capillary electrophoresis condition conducting; S3, mass spectrum condition conducting; S4, quantitative analysis, wherein according to the concentration and the peak area of a standard solution, linear regression is carried out, a linear equation is obtained, and then according to the liner equation and the peak area obtained through detection, the concentration of the phosphatidylcholine and theconcentration of lysophosphatidylcholine in the serum sample are calculated. The application of the method is the application to detecting alzheimer disease. Relative quantitative analysis is carriedout on the PC / PLC in the serum in a multistage reaction monitoring mode (MRM), a large quantity of interfering ions are excluded, the detection limit is 0.0001 ng / mu L, the reproducibility is good, and the recycling rate and the detection sensitivity are high.
Description
technical field [0001] The invention belongs to the technical field of quantitative analysis, and more specifically relates to a method for quantitatively analyzing phosphatidylcholine in serum by CE-MS, and also relates to a method for quantitatively analyzing phosphatidylcholine in serum by capillary electrophoresis coupled with mass spectrometry (CE-MS). Use of the alkaline method. Background technique [0002] Alzheimer's disease (Alzheimer disease, AD), also known as Alzheimer's disease, senile dementia, senile dementia, dementia, etc., is a persistent neurological dysfunction and the most common form of dementia. cause. The most prominent symptom early on is amnesia, usually manifested by progressive short-term memory loss, while long-term memory is relatively unaffected by the condition. As the disease worsens, the patient's language ability, spatial discrimination ability, and cognitive ability will gradually decline. Alzheimer's disease is mainly divided into fam...
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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62G01N27/447
Inventor 胡锐杨运煌张许刘买利
Owner WUHAN INST OF PHYSICS & MATHEMATICS CHINESE ACADEMY OF SCI
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