How to measure heparin sodium
As an anticoagulant substance, heparin sodium usually has a greater anticoagulant effect after being formulated into a solution, but sometimes it cannot be added at will, and heparin sodium needs to be determined according to the calculation formula to ensure the accuracy of dosage sex. According to the experimental requirements, the adsorption rate, absorbance, solution concentration and titer of heparin sodium are generally measured. The following will focus on introducing it to you.
1. Determination of the adsorption rate of heparin sodium
The potency of the heparin sodium solution after adsorption is detected after a certain interval of time, and the total potency remaining in the solution is calculated at the same time, and the adsorption capacity is characterized by the total potency remaining. The titer of heparin sodium solution determined before adsorption was set as D0, and the titer of the experimental solution determined after adsorption was set as D1. (D0-D1) is the D value of resin-adsorbed heparin sodium. The average value of three parallel experiments is taken, and the determination formula is M=(D0-D1)/D0.
2. Determination of absorbance of heparin sodium
Determination of the absorbance value of heparin sodium can adopt ultraviolet spectrophotometer, when the monochromatic light radiation passes through the measured substance solution, it is directly proportional to the amount absorbed by the substance and the thickness and concentration of the liquid layer, and its determination formula is A=lg(1 /T)=E×C×L.
3. Determination of the concentration of heparin sodium solution
Prepare the test solution and the control sample solution during the preparation process. The amount of the tested components contained in the reference solution should be 100% ± 10%, and the reagents used should be consistent. Measure the test solution and the reference substance at the specified wavelength. After measuring the absorbance of the solution, the concentration of the solution can be calculated according to the determination formula.
4. Determination of Heparin Sodium Potency
Usually, the sheep plasma method is used for determination. The water temperature is adjusted to about 37 degrees, and the frozen sheep plasma is taken out, melted and filtered, and set aside. Then two groups of test tubes with different ul were taken respectively, and 8% sodium citrate solution, 0.25% calcium chloride and filtered sheep plasma were added into the tubes. Cover the test tube, invert and mix slowly 3-4 times to make the inner wall wet, put it vertically in a water bath with a constant temperature of 37°C, take out the test tube after 1 hour, check the degree of coagulation, and record and calculate.
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