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Protamine sulfate titer measuring method

A technology of protamine sulfate and a determination method, which is applied in the direction of transmittance measurement and the like, can solve the problems of high inspection cost, complicated operation, long time consumption, etc., and achieves the effect of less experimental reagents and low experimental cost.

Active Publication Date: 2014-02-26
QINGDAO JIULONG BIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The rabbit plasma used in the test is either prepared by yourself or purchased at an expensive price. The price of plasma on the market is about 10 yuan per tube, each tube is 0.5ml, and a sample of protamine sulfate is measured, repeated 5 times, at least 60 Rabbit plasma costs more than 600 yuan, and the cost of testing is very high
In addition, the operation is complex and time-consuming

Method used

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  • Protamine sulfate titer measuring method
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Examples

Experimental program
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Effect test

Embodiment 1

[0018] (1) Preparation of heparin standard solution

[0019] Weigh 150.05mg of heparin standard substance, unit potency 180iu / mg, add 0.9% sodium chloride solution to dissolve to make it 25ml each of heparin standard substance dilutions with different potency concentrations, potency concentrations are 145iu / ml, 150iu / ml respectively ml, 155iu / ml, 160iu / ml, 165iu / ml, 170iu / ml.

[0020] (2) preparation of protamine sulfate solution

[0021] Weigh 122.52 mg of protamine sulfate, batch number 110901, add 0.9% sodium chloride solution, and dilute to a 1 mg / ml solution.

[0022] (3) Test method

[0023] Take 6 test tubes, add 5ml of protamine sulfate solution to each test tube, and then add 5ml to each test tube, the corresponding concentration is 145iu / ml, 150iu / ml, 155iu / ml, 160iu / ml, 165iu / ml, 170iu / ml heparin standard diluent, let it stand, take the supernatant, measure the light transmittance of each tube of the mixed solution at a wavelength of 500nm, and repeat 3 times. ...

Embodiment 2

[0031] (1) Preparation of heparin standard solution

[0032] Take by weighing 152.35mg of heparin standard substance, unit potency 180iu / mg, add 0.9% sodium chloride solution to dissolve and make it become each 25ml of heparin standard substance dilution of different potency concentration, concentration is respectively 145iu / ml, 150iu / ml, 155iu / ml, 160iu / ml, 165iu / ml, 170iu / ml.

[0033] (2) preparation of protamine sulfate solution

[0034] Weigh 125.30 mg of protamine sulfate, batch number 110902, add 0.9% sodium chloride solution, and dilute to a 1 mg / ml solution.

[0035] (3) Test method

[0036] Get 6 test tubes, add 5ml of protamine sulfate solution in each test tube, and then add 5ml in each test tube respectively, the corresponding concentration is 145iu / ml, 150iu / ml, 155iu / ml, 160iu / ml, 165iu / ml, 170iu / ml heparin standard dilution, let it stand, take the supernatant, measure the light transmittance of the mixed solution in each test tube at a wavelength of 450nm, re...

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Abstract

"Chinese pharmacopoeia" (2010 edition) regulates that protamine sulfate titer measurement should adopt bioassay methods. A titer of a sample is measured by comparing the clotting times of fresh rabbit bloods, pig plasmas, or rabbit plasmas which are individually added with a heparin standard substance (S) and the sample (T). The market price of rabbit plasma is high, one dose of rabbit plasma costs 10 yuan, and contains 0.5ml of rabbit plasma; and at least 60 doses are needed to measure a protamine sulfate sample. An ultraviolet spectrophotometer is adopted, the solution light-transmittance is measured under a light with a wavelength of 550 nm, then the protamine sulfate titer is measured, in the operation rabbit plasma is not needed, thus the cost is reduced, and moreover the operation is more convenient.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the determination of protamine sulfate potency. Background technique [0002] "Chinese Pharmacopoeia" 2010 Edition Appendix VII J Protamine Sulfate Bioassay Method, details the determination method of protamine sulfate potency, and its potency is determined by the coagulation status of rabbit plasma. The specific operation method is as follows: [0003] (1) Preparation of heparin standard solution Weigh an appropriate amount of heparin standard, add 0.9% sodium chloride solution to dissolve according to the potency, and make several solutions with different concentrations. [0004] (2) Preparation of protamine sulfate solution Weigh protamine sulfate and add 0.9% sodium chloride solution to prepare a 1 mg / ml solution. [0005] (3) Test method Take 8 small test tubes, the first tube and the eighth tube are blank control tubes, add 0.2ml of 0.9% sodium chloride solution, the second to ...

Claims

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Application Information

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IPC IPC(8): G01N21/59
Inventor 刘乃山葛翠凤刘翠珍孙延年刘振海
Owner QINGDAO JIULONG BIO PHARMA
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