JOURNAL OF CHEMICAL EDUCATION
A MICRO PORPHYRIN TEST FOR BLOOD1 EDITH JU-HWA CHU and T. C. CHU Immaculate Heart College, Los Angeles, California
ALTHOUGH it has been a well-known fact since 18712 that hemoglobin can be converted to protoporphyrin which is easily detected by its red fluorescence under ultraviolet light, this reaction has only recently been employed in qualitative and quantitative clinical worka and has not been included in any laboratory manual of biochemistry. For the latter purpose this report has been written. When a blood sample is boiled with a solution of hydrazine dihydrochloride in glacial acetic acid, the
'
The work was supported by a research grant from The National Institutes of Health, Public Health Service. HOPPE-SEYLER, F., Medizin-ehem. Untemuch~mga,Nos. 1 4 (1871). "BRUGSCH,
J., 2.ges. Inn. Med., 2,191,454 (1947).
former reagent converts hemoglobin into porphyrin and the latter keeps proteins in solution, and therefore the red fluorescence is easily observed. .! he rrngrnr was prrp:~~.riIhy diusolvil~gI g. of hydra-
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1,f zinr r l i l ~ ~ ~ l r ~ ~ c in l ~ al ~n~ixrur? ~ r i i l r conia~ninr5- 0- ml. ~--- - glacial acetic acid and 50 ml. of 0.1 N hydrochloric acid. A blood sample of 0.01 ml. mas taken by a Thomas type blood-diluting pipet and diluted to 50 ml. in a volumetric flask. This solution had a dilution 1:5,000 and was used as a stock solution for preparing other dilutions as follows: 1 : 10,000; 1 :20,000; 1:30,000; and 1:50,000. Samples of 0.1 ml. of each dilution were placed in test tubes of 75 X 12 mm. and respectively mixed with 6 drops of the reagent. They were heated in a boiling water bath for 3 minutes and then
APRIL, 1953
viewed under =mineral light of 3660 k. side by side with a control containing only - the reaeent. From severa1 blood samples it was found that O:i ml. of the dilution as low as 1 :20,000 gave a visible red fluorescence. The sensitivity of the porphyrin test for blood as found
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above, 1:200,000, is comparable with other tests4 and should be useful in the detection of blood stains. ,GRODSKY, M.,K, WRI(IHT, AND ?, L.KIRK, C7imidLaw,
~ ~ (1951).
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~police i xSei., ~ ~~~~h~~~~~ ~ b univ., ~ ~42, 95-104
