Use of dihydroxyglutaric acid or its detection reagent
A technology of dihydroxyglutaric acid and detection reagents, which is applied in the medical field and can solve problems such as inability to predict and stratify
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preparation example Construction
[0094] Preparation of internal standard solution and methoxamine solution
[0095] 1. Weigh a sufficient amount of internal standard 2-chlorophenylalanine powder, add an accurate volume of ultrapure water, oscillate to fully dissolve, then make a 0.3mg / mL aqueous solution, put it in a glass tube, and keep it away from light , and stored in a 4°C refrigerator for later use.
[0096] 2. Weigh a sufficient amount of internal standard heptadecanoic acid powder, add an accurate volume of methanol solution (chromatographically pure grade), oscillate to fully dissolve, and make a 1mg / mL alcohol solution, put it in a glass tube, and keep it away from light , stored at 4°C for later use.
[0097] 3. Weigh an appropriate amount of methoxyamine powder, add an accurate volume of pyridine, shake to dissolve it fully, and make a 15 mg / mL solution, and store it temporarily in the dark at room temperature for later use.
[0098] serum derivatization
[0099] Remove frozen serum from -80°C ...
Embodiment 1
[0112] Embodiment 1 sample collection
[0113] 1. Research object
[0114] 1.1. Experimental example: This study included seven major blood disease diagnosis and treatment centers across the country from 2007 to 2012, including Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, the First Affiliated Hospital of Zhejiang University School of Medicine, and the First Affiliated Hospital of Soochow University. Acute myeloid leukemia confirmed by morphological, immunological and genetic (MIC) examinations Patients, a total of 367 cases, as follows:
[0115] There were 210 males and 157 females. The median age of AML patients was 46 years old, and the age distribution ranged from 15 to 82 years old.
[0116] In addition, other common blood system tumor patients were collected: 150 cases of acute lymphoblastic leukemia, 92 males and 58 females, the median age of the patients was 36 years old, and the age distribution was 13-73 years old; 7 cases of prima...
Embodiment 2
[0123] The configuration of embodiment 2 standard solution and the establishment of standard curve
[0124] Prepare the mother solution of 2-HG standard, and then take an appropriate amount of the mother solution and dilute it into 14 concentrations with different concentrations, and take 100 μL of the diluted solution into the GC high recovery sample bottle, add the internal standard chlorphenylalanine and 10 μL each of heptadecanoic acid was derivatized according to the serum derivatization method, and finally loaded for detection.
[0125] According to the solubility gradient of the standard substance and the corresponding peak area, the regression line of the peak area corresponding to the different solubility of the standard substance is calculated, and the solubility of 2-HG of each sample is determined by the regression line. In order to make the solubility of 2-HG tend to a normal distribution, the 2-HG solubility of all samples was transformed by log2. The converted ...
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