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Urine treatment method and method for detection of methylmalonic acid in urine by gas chromatography-mass spectrometry

A technique of urine methylmalonic acid and gas chromatography, applied in the directions of measuring devices, instruments, scientific instruments, etc., can solve the problems of difficult diagnosis, nerve damage, delayed treatment timing, etc., and achieves simple and easy operation and high accuracy and accuracy, the effect of saving sample processing time

Active Publication Date: 2017-03-01
深圳爱湾医学检验实验室
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  • Summary
  • 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 detection of methylmalonic acid in urine by gas chromatography-mass spectrometry
  • Urine treatment method and method for detection of methylmalonic acid in urine by gas chromatography-mass spectrometry
  • Urine treatment method and method for detection of methylmalonic acid in urine by gas chromatography-mass spectrometry

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Embodiment 1

[0050] The processing of embodiment 1 urine

[0051] The urine of normal neonates and children with methylmalonic acidemia was collected, and processed according to the following steps: (1) the creatinine content in the urine was measured by ultraviolet spectrophotometry, and a urine sample equivalent to 0.2 mg of creatinine was taken; 2) Add the internal standard 2-phenyl-3-hydroxypropionic acid (tropic acid) and 2.0ml of frozen absolute ethanol, fully oscillate and mix evenly with an automatic vibrator; (3) Centrifuge at 4000rpm for 5 minutes to remove protein; (4) transfer the supernatant, and blow it with a nitrogen blower until it is completely dry at 60°C; (5) add bis(trimethylsilyl) tris to the dry residue at 70°C Fluoroacetamide (BSTFA): trimethylchlorosilane (TMCS) 99:1 silylating reagent for 30 minutes derivatization reaction. The whole process takes about 1 hour.

Embodiment 2

[0054] Embodiment 2 detects the processed urine

[0055] The urine sample pre-treated as above is 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 instrument 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] Injection port temperature: 280°C;

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

[0062] Flow control mode: constant line speed;

[0063] 2. MS part

[0064] Ion source temperature: 200°C;

[0065] Interface temperature: 280°C;

[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 spectrum is as figure 1 and figure 2 as shown, figure 1 For the results o...

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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 processing method and a urine organic acid detection method, in particular to a urine methylmalonic acid detection method. Background technique [0002] Methylmalonic acidemia (MMA), also known as methylmalonic aciduria, is an autosomal recessive metabolic disease and the most common type of organic acidemia. The cause of the disease is that there is an obstacle in the metabolic process of converting methylmalonyl-CoA to succinyl-CoA, which leads to the accumulation of organic acids such as methylmalonyl-CoA, methylmalonate, and propionic acid in the body, causing Metabolic acidosis can damage the nervous system. In severe cases, it can cause ketoacidosis, hypoglycemia, hyperammonia, hyperglycinemia, etc., and the mortality rate in neonates and infants is very high. [0003] However, due to the complex clinical manifestations and non-specific symptoms of organic acid metabolic diseases, clinical diagnosis is difficult,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88G01N30/06
Inventor 杨江涛钱俊
Owner 深圳爱湾医学检验实验室