Preparation method based on hemoglobin and four-carbon or five-carbon dibasic acid complex kit
A technology of hemoglobin and dibasic acid is applied in material analysis by observing the influence on chemical indicators, analysis by chemical reaction of materials, color/spectral property measurement, etc., which can solve the problems of low accuracy and complicated operation. , toxic pollution and other problems, to achieve the effect of low cost, high sensitivity and low price
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Embodiment 1
[0026] Preparation of hemoglobin-tartaric acid complex.
[0027] Add 100 μL 0.03M tartaric acid to 100 μL 1mg / mL hemoglobin, and mix well to obtain the hemoglobin-tartaric acid complex, whose circular dichroism diagram is as follows figure 1 As shown, after the addition of tartaric acid, the secondary structure of hemoglobin changed, binding figure 2 , image 3 , the catalytic activity of hemoglobin added with tartaric acid was significantly enhanced, and the absorbance of iron oxide TMB at 652nm was significantly increased. Hemoglobin was heated at different temperatures for 1 hour and tartaric acid was added, the catalytic performance of the complex was not significantly weakened, as shown in Figure 4 , Figure 5 , Figure 6 as shown, Figure 5 It is the ultraviolet absorption spectrum of the hemoglobin and tartaric acid complex catalytically decomposing hydrogen peroxide and oxidizing TMB to generate ox-TMB at different temperatures. Because high temperature has litt...
Embodiment 2
[0029] The hemoglobin-glutaric acid complex is used for the detection of hemoglobin in blood.
[0030] Prepare a microplate with standard wells and sample wells. Add 50 μL standard (different concentrations of hemoglobin and glutaric acid complexes) and 50 μL samples (serum diluted 10 times) to the microwell plate, 50 μL 2.5mM H 2 o 2 , 50μL 2.5mMTMB, and measure the absorbance at 652nm after 10 minutes. The final concentration of the standard is 0.01g / L, 0.02g / L, 0.04g / L, 0.06g / L, 0.08g / L, 0.1g / L. Take the concentration of the standard substance as the abscissa and the absorbance as the ordinate to make a standard curve, such as Figure 8 shown. Then, the absorbance of each sample group was substituted into the standard curve to obtain the hemoglobin concentration, and the results are shown in Table 1.
[0031] Table 1
[0032] Sample serial number 1 2 3 4 5 6 7 8 9 Elisa reference value (g / L) 0.21 0.341 0.255 0.385 0.3125 0.3605 0.341 0....
Embodiment 3
[0034] The hemoglobin-tartaric acid complex is used for the detection of glucose in the blood.
[0035]Prepare 12 centrifuge tubes, numbered 1-12, and centrifuge tube No. 1 is the control group. Add 100 μL of 11 different concentrations of glucose solutions to centrifuge tubes 2-12, and then add 100 μL of glucose oxidase with a concentration of 0.5 mg / mL, and put it in a water bath at 37°C for 30 minutes Then add 800 μL of hemoglobin-tartaric acid with a concentration of 0.5 mg / mL and 400 μL of 5 mM TMB to each centrifuge tube, and place it at room temperature for 35 minutes. Finally, measure the absorbance of ox-TMB with a UV-visible spectrophotometer. The final concentrations of glucose were 0.005, 0.01, 0.03, 0.05, 0.07, 0.09, 0.11, 0.13, 0.15, 0.2, 0.25 mM.
[0036] Take the final concentration of glucose as the abscissa and the absorbance of ox-TMB as the ordinate to make a standard curve, such as Figure 9 shown. Prepare 9 more centrifuge tubes, numbered A-I, and No. ...
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