Intended use: This reagent kit is used for in vitro determination of low-density lipoprotein cholesterol (LDL-C) content in human serum.
The main physiological function of low-density lipoprotein cholesterol is to transport phospholipids and cholesterol. It is an anti atherosclerotic lipoprotein and a protective factor of coronary heart disease. The content of low-density lipoprotein cholesterol (LDL-C) is significantly negatively correlated with the degree of arterial stenosis. Clinical analysis of different types of lipoprotein ratios is used as a differential diagnosis for different types of hyperlipidemia.
Elevated low-density lipoprotein cholesterol is commonly found in alcohol consumption, long-term physical activity, and other conditions. The reduction of HDL cholesterol is common in coronary heart disease, cerebrovascular disease, diabetes, hepatitis, cirrhosis, etc.
Verification principle
The surface of chylomicrons (CM), low density lipoprotein (LDL), and extremely low density lipoprotein (VLDL) in serum is masked by polyanions and reaction inhibitors. Under the action of cholesterol enzyme reagents, HDL-C, surfactants, and reaction promoters
Participated in the Trinder reaction for color development, and the absorbance is directly proportional to the concentration of HDL-C in the sample.
Storage conditions and expiration date
The unopened reagents are stored in dark at 2-8 ℃ and have a validity period of 12 months; After the reagent is unsealed, it is stored in a dark place at 2-8 ℃ and is valid for one month without contamination; Reagents should not be frozen.
Sample requirements
1. Use fresh non hemolytic serum
No significant interference was observed when hemoglobin ≤ 500g/L, triglycerides ≤ 10g/L, and ascorbic acid ≤ 1704umol/L were present in the sample.
3. Stability: It can be stable for 3 days when stored at 2-8 ℃. If it cannot be measured in a timely manner on the same day, please store it at -25~-15 ℃. Before use, the sample should be quickly thawed at 37 ℃ and should not be repeatedly frozen or thawed. Do not use contaminated samples.
Applicable instruments
Hitachi 7180/7170/7060/7600 fully automatic biochemical analyzer, Dirui CS-400B, Abbott c16000, OLYMPUS AU640 fully automatic biochemical analyzer.
Inspection method
1. Reagent preparation: Liquid reagent can be used upon activation
2. Measurement conditions: Main wavelength: 546nm; Sub wavelength: 700nm; Color comparison cup optical path: 1cm; Temperature: 37 ℃; Analysis type endpoint method
3. Calibration procedure: Use purified water and high-density lipoprotein cholesterol (HDL-C) calibration samples for two-point calibration. The calibration samples should be purchased separately, and it is recommended to use Randox or other traceable calibration samples. The calibration adopts linear calibration mode.
4. Quality control program: Measure high-density lipoprotein cholesterol (HDL-C) quality control samples, and the test results can only be tested within the quality control range.
Reference range: 0.75~2.26mmol/L
Determination method: Select no less than 200 normal human serum samples through clinical trials, and measure them with a fully automated biochemical analyzer. The measured values are processed using statistical methods and the reference interval is calculated.
It is recommended that each laboratory establish their own reference range!
Limitations of testing methods
Bilirubin< 50 mg/dL, intralipid< 3000 mg/dL, ascorbic acid< 50 mg/dL, hemoglobin< 500mg/dL will not interfere with the determination of HDL-C in the sample. Severe lipidosis and hemolysis have an impact on the measurement results. The components in different batch number kits cannot be exchanged or mixed for use.
Interpretation of inspection results
Human errors, sample processing, and deviations from analytical instruments can all affect the measurement results; When individual samples deviate too far from the expected value, it is necessary to retest.
Product performance indicators
1. Under the conditions of 37 ℃, 546nm wavelength, and 1 cm optical path, the blank absorbance of the reagent shall not exceed 0.03.
2. Precision: The repeatability CV shall not exceed 3%; The relative deviation R between batches shall not exceed 5%.
3. Accuracy: For testing traceability standards, the absolute deviation within the range of [0.2~1.0] mmol/L should not exceed ± 0.25mmol/L, and the relative deviation should not exceed ± 10% (1.0~6.0] mmol/L)
4. Linear range:
1) Within the range of 0.15 to 6.00mmol/L, the correlation coefficient r should not be less than 0.990;
2) Within the range of [0.15~2.50] mmol/L, the absolute deviation between the measured concentration and the estimated value should not exceed ± 0.25mmol/L; Within the range of 2.50-6.00] mmol/L, the relative deviation between the measured concentration and the estimated value should not exceed ± 10%.
5. Analysis sensitivity: When testing 1.00mmol/L high-density lipoprotein cholesterol, the absolute value of absorbance difference should not be less than 0.04.
6. Stability: All dosage forms of reagents can be used before the expiration date indicated on the label under strict conditions of avoiding light and storing at 2-8 ℃; The reagent can remain stable for 30 days after opening the bottle.
matters needing attention
1. Avoid contact with skin and mucous membranes with reagents. Necessary preventive measures should be taken when using reagents. If the reagents come into contact with the skin and mucous membranes, rinse them with water and seek medical attention if necessary.
2. The upper linear limit of this method is 6.0mmol/L. If the measured value of the sample exceeds the upper limit, dilute it with 0.9% sodium chloride solution before determining, and multiply the result by the dilution factor.
3. Double reagent method should be used for operation, and single reagent operation should not be used.
4. Waste liquid treatment: It is recommended to refer to the requirements of local regulations.
5. It is recommended that each laboratory verify other models of instruments themselves. If you need detailed measurement parameters, please contact our company.
6. Different batch numbers of reagents cannot be mixed. When changing the batch number of reagents, please recalibrate!
