Use Guide for Heparinases

Published on 2015/1/5 15:20:11

A heparinase kit contains 0.1 IU of heparinase I, 0.1 IU of heparinse II and 0.1 IU of heparinase III. Here is the instruction for the test of 1,6-anhydro forms in enoxaparin sodium.

Add potassium phosphate pH 7.0 buffer into the tubes as the following table.

Enzyme Buffer Added Concentration
Heparinase I, 0.1 IU, 10 uL 240 uL 0.4 IU/mL
Heparinase II, 0.1 IU, 25 uL 225 uL 0.4 IU/mL
Heparinase III, 0.1 IU, 20 uL 230 uL 0.4 IU/mL

Store these diluted heparinases at -20 ℃ until use.

Before digesting your enoxaparin samples, pool the three diluted heparinases together.

Pipette 100 uL of the mixture of heparinases, 20 uL of test solution or stardand solution, 70 uL of sodium/calcium acetate pH 7 solution. Mix gently, then incubate the reaction solution at 25 ℃ in a water bath for at least 48 hours.

The two aforementioned buffers are preprepared according to USP chapter <207>.