Application
The kit is used for the quantitative determination of the total bile acid concentration in serum.
Detection significance
Total bile acid (TBA) is mainly used for the screening and prognosis of follow-up of hepatobiliary disease and as the marker of liver parenchymal damage and cholestasis. The increase of TBA indicates the risk of viral hepatitis, cirrhosis, alcoholic liver disease, drug-induced liver injury or cholestasis.
Detection principle
Measure the OD value at 405 nm and the changes of absorbance is proportional to the concentration of bile acid.
Sample requirements
1. Separate serum within 2 hours after blood collection. The serum sample can be stored at 15~30℃ within 8 hours, at 2~8℃ for a week or at -20℃ for 3 months.
2. Interfering substances: conjugated bilirubin≤ 5mg/dL, unconjugated bilirubin ≤ 20mg/dL, vitamin C ≤ 1mg/dL, glycerin trilaurate ≤ 9.25 mmol/L, hemoglobin ≤ 100mg/dL have no effect to the results.
Operation steps
1. Detection with Spectrophotometer
Operation table
| Standard well | Sample well |
Standard (μL) | 12 |
|
Sample (μL) |
| 12 |
Reagent 1 (μL) | 800 | 800 |
Reagent 2 (μL) | 200 | 200 |
Mix fully and incubate at 37℃ for 1.5 min. Measure the absorbance at 405 nm at 0 second (A1) and 2 min (A2), respectively. Calculate the △A/min. |
2. Detection with Microplate reader
Operation table
| Standard well | Sample well |
Standard (μL) | 3 |
|
Sample (μL) |
| 3 |
Reagent 1 (μL) | 200 | 200 |
Reagent 2 (μL) | 50 | 50 |
Mix fully and incubate at 37℃ for 1.5 min. Measure the absorbance at 405 nm at 0 second (A1) and 2 min (A2), respectively. Calculate the △A/min. |
3. Detection with Biochemical analyzer
Setting parameter
Temperature | 37℃ | Method | Two-point end point method |
Dominant wavelength | 405 nm | Optical path | 1 cm |
Reaction direction | Up | Sample | 3 μL |
Reagent 1 | 200 μL | Reagent 2 | 50 μL |
Reaction time | 2 min |
|
|
Measure the absorbance at 0 second (A1) and 120 second (A2), respectively. Calculate the △A/min.
Automatic biochemical analyzer has its own program parameter input language. Reagents matches the analyzer and carry out automatic measurement after the above basic parameters are modified.
Reference range
1.2~10.5 μmol/L (This data is for reference only. It is recommended that each laboratory establish its own range of reference values.)
Technical parameter
1. Linear range: 0 ~ 150 μmol/L, r2 ≥ 0.990.
2. Accuracy: inaccuracy ≤ 15.0%.
3. Recovery rate: 100 ± 20%
4. Precision: intra-CV ≤ 5.0%, inter-CV ≤ 10.0%.
5. Absorbance for the blank control (reagents only) ≤ 0.7 (405 nm wavelength, 1 cm optical path).
Notes
1. Instructions should be followed strictly, changes of operation may result in unreliable results.
2. The validity of kit is 12 months.
3. Do not use components from different batches of kit.
4. The sample needs to be diluted with normal saline before the determination when the concentration of TBA is higher than 150 μmol/L. The result should be multiplied by the dilution factor.
5. The kit is for research use only and contains preservatives. It should be avoided to contact with skin and clothing. Wash immediately with plenty of water if contact it carelessly.
6. The ratio of sample and reagent can be scaled as required.
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