Application
This kit can be used for detection of blood ammonia content in serum samples.
Detection significance
Ammonia produced by the catabolism of various amino acids in various tissues of the body and ammonia absorbed by the intestine may enter the blood to form blood ammonia. Normally, ammonia can be transformed into urea in the liver. Serious liver disease results in damage to hepatic parenchymal cells. Ammonia cannot be removed from the circulatory system, resulting in increased blood ammonia content.
Detection principle
Blood protein can be precipitated with protein precipitator, and enzyme activity will be destroyed, which can prevent the formation of free ammonia in vitro. Most interfering color substances were removed at the same time, indigo was formed in non-protein filtrate by Berthelot reaction, and the color depth was proportional to the content of blood ammonia. Blood ammonia content can be determined by comparing with standard solution.
Experimental instrument
Tube, Micropipettor, Vortex mixer, 37℃ water bath, Centrifuge, Spectrophotometer (635 nm)
Sample preparation
Serum Sample: Detect the sample directly.
Note: hemolysis and turbid serum may affect the result. If there are turbidity or flocculent precipitates in the serum samples, the sample should be centrifuged and then take the supernatant for detection.
Operation steps
1. Pretreatment of samples
| Blank tube | Standard tube | Sample tube |
Standard diluent (mL) | 0.2 |
|
|
350 μmol/L Standard working solution (mL) |
| 0.2 |
|
Sample (mL) |
|
| 0.2 |
Reagent 1 | 0.6 | 0.6 | 0.6 |
Mix fully and centrifuge at 3500 rpm for 10 min, then take the supernatant for chromogenic reaction. |
2. Chromogenic reaction
| Blank tube | Standard tube | Sample tube |
Supernatant (mL) | 0.4 | 0.4 | 0.4 |
Reagent 2 (mL) | 1.0 | 1.0 | 1.0 |
Reagent 3 (mL) | 1.0 | 1.0 | 1.0 |
Mix fully and incubate at 37℃ for 30 min. Set to zero with double-distilled water and measure the OD values of each tube at 635 nm with 1 cm cuvette. |
Technical parameter
1. The sensitivity of the kit is 10.4 μmol/L.
2. The intra-assay CV is 4.7% and the inter-assay CV is 5%.
3. The recovery of the kit is 103%.
4. The detection range of the kit is 0.0104-2.0 mmol/L.
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