Nicotinamide agent and preparation method thereof
A nicotinamide and reagent technology, applied in the field of reduced nicotinamide coenzyme assay reagents, can solve the problems of difficult enzyme content determination and the like
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Embodiment 1
[0113] Embodiment 1: prepare alanine nitrogen base transferase reagent, be made up of the following components by the content of every liter,
[0114] Tris buffer 100mM pH7.15, 37°C,
[0115] L~alanine 500mM,
[0116] Lactate dehydrogenase 2.7ku / L,
[0117] α-ketoglutaric acid 15mM,
[0118] EDTA.Na 2 .2H 2 O 60mM
[0119] NAD 0.27mM
[0120] D~glucose 50mM.
[0121] Glucose-6-phosphate dehydrogenase (Leuconostoc) 3700U / L,
[0122] Dipotassium hydrogen phosphate 5mM
[0124] According to the method of the above reagent solution, after the preparation of the above alanine aminotransferase reagent is completed, place it at 2-8°C for 1-3 days, and the reagent blank absorbance is ≥1.10, 340nm, 10mm optical path. When measuring the sample, use the rate method, the sample reagent ratio is 1:12, the temperature is 37°C, the delay time is 90 seconds, the measurement time is 180 seconds, and the number of readings is ≥ 6 points.
Embodiment 2
[0125] Embodiment 2: preparation aspartate aminotransferase reagent, is made up of the following components by the content of every liter,
[0126] Tris buffer 80mM, pH7.50, 37°C,
[0127] L~aspartic acid 240mM,
[0128] Malate dehydrogenase 600U / L
[0129] Lactate dehydrogenase 1000U / L
[0130] a~Ketoglutarate 12mM,
[0131] EDTA.Na 2 .2H 2 O 6mM
[0132] NAD 0.28mM
[0133] D~glucose 80mM.
[0134] Glucose-6-phosphate dehydrogenase (Leuconostoc) 3700U / L,
[0135] Dipotassium hydrogen phosphate 5mM
[0137] According to the above method for preparing the reagent solution, after the preparation of the above aspartate aminotransferase reagent is completed, place it at 2-8°C for 1-5 days, and the reagent blank absorbance is ≥1.20, 340nm, 10mm optical path. When measuring the sample, use the rate method, the sample reagent ratio is 1:12, the temperature is 37°C, the delay time is 90 seconds, the measurement time is 180 seconds, and the numbe...
Embodiment 3
[0138] Embodiment 3: prepare urea reagent, be made up of following component by the content of every liter,
[0139] Tris buffer pH7.50, 37°C, 14.54g
[0140] Alpha-ketoglutarate 1.43g
[0141] Bovine Serum Acid 5.0g
[0142] Adenosine diphosphate potassium salt 0.65
[0143] Glutamate dehydrogenase (microbial source) 550u
[0144] Urease 12000u
[0145]NAD 0.192g
[0146] D~glucose 18.0g
[0147] Glucose-6-phosphate dehydrogenase (Leuconostoc) 3700u
[0148] Dipotassium hydrogen phosphate 0.87g
[0149] EDTA.Na 2 .2H 2 O 0.37g
[0151] According to the method for preparing the reagent solution above, after the preparation of the above urea reagent is completed, place it at 2-8°C for 1-7 days, and the reagent blank absorbance is ≥1.20, 340nm, 10mM optical path. When measuring the sample or calibrator, use the two-point method, the sample reagent ratio is 1:100, the temperature is 37°C, the delay time is 30 seconds, the measurement time i...
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