Glycosylated hemoglobin reference material and preparation method thereof
A technology of glycosylated hemoglobin and standard substances, which is applied in the field of medical testing, can solve the problems of lack of HbA, etc., and achieve the effects of convenient transportation, simple preparation method, and convenient use
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Embodiment 1
[0048] Embodiment 1: Preparation of glycosylated hemoglobin standard substance
[0049] 1. Materials
[0050] 1. Selection of packaging containers
[0051] The polyethylene material does not absorb and release the components in the candidate standard substance matrix, and is heat-resistant, acid-resistant, alkali-resistant, and water-resistant. Therefore, a sterile polyethylene tube (Axygen Company, USA) was selected as the standard substance container.
[0052] 2. Disinfection
[0053] Before preparation, the aseptic room and ultra-clean bench were sterilized by ultraviolet light.
[0054] 3. Selection of volunteers for blood collection
[0055] Healthy volunteers without diabetes, well-controlled diabetes, and poorly-controlled diabetes, aged 20 to 50 years, half male and half female, negative for human immunodeficiency virus antibody (HIV) and hepatitis B and C surface antigen (HBV, HCV), fill in Informed Consent.
[0056] 4. Reagents
[0057] Acetonitrile, chromatogr...
Embodiment 2
[0070] Example 2: Uniformity Test
[0071] 1. Detection method
[0072] Ion-exchange HPLC method is commonly used in clinical HbA 1c One of the measurement methods, the precision of this method is good, it can meet the requirements of uniformity test, and the detection cost is low, the time is short, and it is suitable for the analysis of large sample size. Therefore, after comprehensive consideration, the research unit adopts the ion-exchange HPLC method as the determination method for the homogeneity test.
[0073] 1. Adopt the random sampling method, number the candidate standard substance that subpackaging embodiment 1 obtains sequentially, each concentration randomly selects 21 bottles to carry out homogeneity test, the method for measuring is ion-exchange HPLC method, and the minimum sampling volume is 5 μ l. All samples were stored at -70°C until assayed.
[0074] 2. Before the measurement, the sample was reconstituted at room temperature, and vortexed on a blood mix...
Embodiment 3
[0100] Embodiment 3: stability test
[0101] 1. Detection method
[0102] According to the description in GB / T15000.3-2008 / ISO Guide35:2006, the stability inspection research is carried out by using the synchronous stability research. For the short-term stability study, a total of 9 temperature time points were designed, which were zero point, 1 day, 2 days, and 4 days at room temperature; 1 week, 2 weeks, and 1 month at 4°C; 2 weeks, 1 month at -20°C and 3 months, 5 temperature time points are designed for long-term stability research, which are zero point, -70 ℃ 1 month, 3 months, 6 months, 9 months and 12 months; Bottle samples were analyzed in triplicate for each bottle. Same as the homogeneity test study, the stability test uses the ion-exchange HPLC method as the analytical method.
[0103] 1. Sample storage for stability testing studies
[0104] The samples at each temperature and time point were randomly selected from the total sample as follows:
[0105] 3 bottle...
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