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Urine treatment method and method for detecting urine methylmalonic acid through gas chromatography and mass spectrometry

A treatment method, gas chromatography technology, applied in the field of urine treatment methods and urine organic acids, can solve problems such as difficult diagnosis, nerve damage, delay in treatment timing, etc., and achieve simple and easy operation, high precision and accuracy, The effect of saving sample processing time

Active Publication Date: 2015-10-21
深圳爱湾医学检验实验室
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0020] In summary, in view of the high incidence of methylmalonic acidemia in organic acid metabolism diseases, which will cause irreversible neurological damage and other serious consequences in children, the sensitivity index of tandem mass spectrometry filter paper dry blood spot detection has the same relationship with propionic acidemia. Similarly, the specificity is not high, and it is not easy to diagnose; as the current main detection method, GC-MS combined with urine organic acid detection takes a long time, which may delay the best time for treatment

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  • Urine treatment method and method for detecting urine methylmalonic acid through gas chromatography and mass spectrometry
  • Urine treatment method and method for detecting urine methylmalonic acid through gas chromatography and mass spectrometry
  • Urine treatment method and method for detecting urine methylmalonic acid through gas chromatography and mass spectrometry

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Embodiment 1

[0050] The treatment of embodiment 1 urine

[0051] The urine of normal newborns and children with methylmalonic acidemia was collected and processed according to the following steps: (1) Measure the creatinine content in the urine by UV spectrophotometry, and take a urine sample equivalent to 0.2 mg of creatinine; ( 2) Add the internal standard 2-phenyl-3-hydroxypropionic acid (tropine acid) and 2.0 ml of frozen absolute ethanol, fully shake and mix with an automatic shaker; (3) use a centrifuge at 4000 rpm for 5 minutes to remove protein; (4) transfer the supernatant to complete dryness with a nitrogen blower at 60°C; (5) add bis(trimethylsilyl)trisium to the dried residue at 70°C Fluoroacetamide (BSTFA):trimethylchlorosilane (TMCS) 99:1 silylation reagent was derivatized for 30 minutes. The entire process takes about 1 hour.

Embodiment 2

[0054] Embodiment 2 detects processed urine

[0055] The urine samples pretreated as above are respectively detected by the existing GC-MS urine organic acid detection method and the GC-MS urine methylmalonic acid detection method of the present invention.

[0056] The instrumental conditions of the existing GC-MS urine organic acid detection method are as follows:

[0057] 1.GC part

[0058] Injection mode: split mode;

[0059] Split ratio: 10:1;

[0060] Inlet temperature: 280℃;

[0061] Heating program: hold 100°C for 4 minutes, ramp up to 280°C at a ramp rate of 4°C / min, hold for 11 minutes

[0062] Flow control mode: constant linear velocity;

[0063] 2. MS part

[0064] Ion source temperature: 200℃;

[0065] Interface temperature: 280℃;

[0066] Sampling method: Scan;

[0067] Interval: 0.50s;

[0068] Scan speed: 1000;

[0069] start m / z: 50;

[0070] End m / z: 500.

[0071] The detected pattern is as figure 1 and figure 2 shown, figure 1 For normal neonat...

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Abstract

The invention discloses a urine treatment method. The urine treatment method comprises the following steps that a certain number of urine samples are taken, an interior label and frozen absolute ethyl alcohol are added, the mixture is fully oscillated and mixed uniformly, centrifugal treatment is carried out for removing protein, supernatant liquor is taken, nitrogen blowing is carried out until the supernatant liquor is fully dried, and derivatization reagent is added for reaction. The invention further discloses a method for detecting urine methylmalonic acid through gas chromatography and mass spectrometry. The method includes the urine treatment method and optimizes the instrument condition of gas chromatography and mass spectrometry detection after the treatment. By the adoption of the urine treatment and detection method, time for urine sample treatment is greatly saved, and organic acid, especially methylmalonic acid can be detected easily, fast and accurately.

Description

technical field [0001] The invention relates to a urine treatment method and a urine organic acid, especially a detection method of urine methylmalonic acid. Background technique [0002] Methylmalonic acidemia (MMA), also known as methylmalonic aciduria, is an autosomal recessive metabolic disorder and the most common type of organic acidemia. The cause of the disease is due to obstacles in the metabolic process of the conversion of methylmalonyl-CoA to succinyl-CoA, which leads to the accumulation of organic acids such as methylmalonyl-CoA, methylmalonic acid, and propionic acid in the body, causing Metabolic acidosis damages the nervous system. In severe cases, it can cause ketoacidosis, hypoglycemia, hyperammonemia, hyperglycinemia, etc. The mortality rate of neonates and infants is very high. [0003] However, due to the complex clinical manifestations and non-specific symptoms of organic acid metabolism diseases, clinical diagnosis is difficult, and mass spectrometry ...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
Inventor 杨江涛
Owner 深圳爱湾医学检验实验室
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