Myoglobin, like hemoglobin, contains the heme gene in the molecule and has peroxidase-like activity that can be detected by the reaction of benzidine and o-toluidine with hydrogen peroxide. Hemoglobin Pr is soluble in 0.8 saturated sulfuric acid solution, while hemoglobin is not, and thus can be identified, this experiment is from the Mudanjiang Medical College undergraduate 5-year clinical laboratory instructions

Operation method

Qualitative urine myoglobin test experiment

Principle

Myoglobin, like hemoglobin, contains the heme gene in the molecule and has peroxidase-like activity that can be detected by the reaction of benzidine and o-toluidine with hydrogen peroxide. Hemoglobin Pr is soluble in 0.8 saturated sulfuric acid solution whereas hemoglobin is not and can be identified.

Materials and Instruments

Urine
o-Toluidine methanol saturated solution Hydrogen peroxide acetic acid solution Amine sulfate powder

Move

I. Experimental reagents:

1, o-toluidine methanol saturated solution, more difficult to dissolve, but very stable.

2, hydrogen peroxide acetic acid solution glacial acetic acid 1 plus 0.9 mol / L hydrogen peroxide solution 2.

3, amine sulfate powder: chemically pure products.

Second, the experimental method:

l, take 1.0 ml of urine, add 0.12 mol / L sulfosalicylic acid 3.0 ml. If turbidity is produced then continue

Otherwise, it means that there is no protein in the urine, and this test is also negative.

2, another 5.0 ml of fresh urine, add 2.8 grams of amine sulfate powder. Mixed to dissolve (about 0.8 saturation) for 5 minutes, filtered with filter paper, still red or brownish-black is positive. Can also take the filtrate according to the hemoglobin qualitative method of operation, such as positive for myoglobin positive.

Caveat

1, the specimen must be fresh, in order to avoid myoglobin denaturation and precipitation by 0.8 saturation ammonium sulfate.

2， this method is a screening test, in a small number of normal human urine may also appear false-positive; further available ultrafiltration method, electrophoresis, chemical identification to identify.

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