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A high-sensitive blood-plasma total homocysteine detecting reagent box

A homocysteine ​​and kit technology, applied in the field of high-sensitivity plasma total homocysteine ​​determination kits, can solve the problems of poor specificity, difficulty in meeting high-level clinical research, high technical requirements, and high reagent prices. Achieve the effect of optimized mobile phase, accurate detection results and easy operation

Inactive Publication Date: 2009-03-04
上海特敏生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Liquid chromatography has the characteristics of specificity and sensitivity, and is a general method for clinical research at present, but its operation is complicated and the technical requirements are high
The operation of immunization method is relatively simple, but the specificity is poor and it is difficult to meet the needs of high-level clinical research, and the price of reagents calculated per person is high

Method used

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  • A high-sensitive blood-plasma total homocysteine detecting reagent box
  • A high-sensitive blood-plasma total homocysteine detecting reagent box
  • A high-sensitive blood-plasma total homocysteine detecting reagent box

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Kit reagent preparation:

[0035] Sample tube: 2 μg of 3-deazaadenosine was added as a stabilizer;

[0036] Reference substance: dialyzed blank serum with a Hcy concentration of 0, adding Hcy chemicals to prepare Hcy concentrations of 45, 15 and 5 μmol / L;

[0037] Quality control: dialyzed blank serum with a Hcy concentration of 0, adding Hcy chemicals to prepare Hcy concentrations of 30 μmol / L and 10 μmol / L;

[0038] Internal standard solution: DL-homocystine-D8 aqueous solution, the concentration is 10 μmol / L;

[0039] Reducing agent: 1,4-dithiothreitol aqueous solution, the concentration is 75mmol / L;

[0040] Protein precipitant: trichloroacetic acid aqueous solution, the concentration is 15% (w / v)

[0041] 2. Blood sample collection: Collect venous blood in a sample tube, centrifuge the plasma (4000rpm×10min) within 8 hours, and store it at room temperature.

[0042]3. Sample processing: Take 50 μl of the plasma to be tested and place it in a centrifuge tube,...

Embodiment 2

[0057] The kit, blood sample collection and pretreatment methods are the same as in Example 1 except that the reducing agent 1,4-dithiothreitol aqueous solution has a concentration of 100 mmol / L, and the mass spectrometry conditions are also the same as in Example 1.

[0058] Chromatographic conditions: Aquasil octadecylsilane bonded silica gel is used as the stationary phase, mobile phase A: 0.1% formic acid in methanol solution, mobile phase B: 0.1% formic acid in water, the ratio is 10:90; isocratic elution, flow rate 0.25mL / min, inject 5μl.

[0059] Under these conditions, the retention time of Hcy and the internal standard are both 1.05 min, the total analysis time of a sample is 2.0 min, and the minimum detection limit is 0.1 μmol / L (signal-to-noise ratio ≥ 5).

[0060] The total homocysteine ​​concentration in the sample was calculated by the peak area or peak height according to the internal standard method.

Embodiment 3

[0062] 2 μg of sodium heparin was added to the sample tube as a stabilizer, and the other kits, blood sample collection and pretreatment methods were the same as in Example 1, and the chromatographic and mass spectrometry conditions were also the same as in Example 1.

[0063] Calculate the total homocysteine ​​concentration in the sample by the peak area or peak height method according to the internal standard method.

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Abstract

The invention relates to a high sensitive blood plasma homocysteine testing kit that includes sample tube with stabilizer, reference substance and quality control serum of different Hcy thickness, DL- homocystine-D8 internal standard solution, reducer and trichloroacetic acid albumen precipitation. The invention has strong specificity for sample testing, and high sensitivity. It could be accurately and rapidly used in Hcy research.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a highly sensitive blood plasma total homocysteine ​​assay kit. Background technique [0002] Homocysteine ​​(Hcy), also known as homocysteine, is an intermediate product of methionine metabolism and has been considered a new independent pathogenic factor by the international medical community. The results of animal experiments have shown that Hcy can induce neural tube defects in embryos, and the accumulation of Hcy in the blood can lead to vascular damage. Existing research results have shown that certain congenital diseases, mental retardation, arterial and venous thrombosis, chronic kidney disease, type II diabetes and Alzheimer's syndrome are often accompanied by abnormal homocysteine ​​metabolism. At present, in developed countries, Hcy has been recognized by the public and has become a routine item of clinical testing. In recent years, clinical research on Hcy in my count...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/72B01J20/281
Inventor 潘俊芳周菊珍刘罡一林益华贾晶莹余琛
Owner 上海特敏生物医药科技有限公司
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