Toluidine blue can neutralize the anticoagulant effect of heparin, when the prothrombin time is prolonged, can be added to the plasma to be examined, or prolonged prothrombin time back to normal means that heparin increases, otherwise other antithrombin-like substances. This experiment is from the Mudanjiang Medical College undergraduate 5-year laboratory guidance for testing majors

Operation method

Plasma heparin or heparin-like anticoagulant screening tests

Principle

Toluidine blue can neutralize the anticoagulant effect of heparin, when the prothrombin time is prolonged, can be added to the plasma to be examined methylamine blue, or prolonged prothrombin time back to normal means that heparin increased, otherwise other antithrombin-like substances.

Materials and Instruments

Toluidine blue solution Sodium citrate solution Thrombin

Move

I. Experimental reagents:

1. 1g/L toluidine blue solution

2. 109mmol/L sodium citrate solution

3. Prothrombin solution same as above prothrombin assay

4. stopwatch

II. Experimental manipulation:

l. Take 0.lml of sodium citrate anticoagulated plasma to be examined and add 10g/L toluidine blue 0.1ml, shake well. Place in a 37℃ water bath and incubate.

2. Add 0.1ml of thrombin immediately, record the coagulation time with a stopwatch, repeat 2-3 times and take the average value.

Experimental results: in patients with prolonged prothrombin time (TT), adding toluidine blue TT shortened by more than 5 seconds, suggesting that the plasma to be examined has heparin or heparin-like increase.

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