APPENDIX 3QI 



Method: Take 20 c.c. of urine, dilute with an equal amount of water and add a 

 few drops of acetic acid. Next boil in a beaker until the original amount of diluted 

 urine is reduced to 10 c.c. (originally 40 c.c.). Dilute this evaporated urine with an 

 equal amount of water, giving us 20 c.c. In each of two test-tubes put 10 c.c. of 

 this 20 c.c. To one tube add i c.c. of hydrogen peroxide and warm gently, without 

 boiling, for one minute; then cool. The other tube is left untreated. Next, to each 

 test-tube add 10 drops of glacial acetic acid and 5 to 10 drops of a freshly prepared 

 sodium nitroprusside solution and mix. Next carefully overlay each tube with 

 about 2 c.c. of concentrated ammonia. If /2-oxybutyric acid were present in the 

 tube treated with the hydrogen peroxide and thereby oxidized to acetone a violet- 

 red ring will develop at the point of contact while in the untreated tube there will 

 be no such color ring. 



A yellowish-brown ring from the presence of creatinin may show in the untreated 

 tube. It is well to allow the tubes to stand for three to four hours before finally 

 reporting the absence of /3-oxybutyric acid. It will probably show 0.2%. 



Acetone. To one-sixth of a test-tube of urine add a crystal of sodium nitro- 

 prusside. Make strongly alkaline with NaOH. Shake. The addition of a few 

 drops of glacial acetic gives a purple color to the foam, if acetone is present. 



Diazo Reaction. To 5 c.c. sulphanilic acid solution (sulphanilic ac. i pt., HC1 

 50 pts., aq. 1000 pts.) add two drops of a 0.5% solution of sodium nitrite. Add an 

 equal quantity (5 c.c.) of urine. Shake and add quickly 2 or 3 c.c. of ammonium 

 hydrate. A carmine color, especially in the foam, shows a diazo reaction. If the 

 reaction is positive, and the mixture is allowed to stand for 24 hours, a precipitate 

 forms, the upper margin of which exhibits a green, greenish-black or violet zone. 



Indican. Take 10 c.c. urine and treat it with i c.c. of sol. of lead subacetate. 

 Filter. Of this nitrate take 6 c.c. and treat with an equal amount of Obermayer's 

 reagent; allow to stand for 5 minutes then shake gently with 2 c.c. of chloroform. 

 Obermayer's reagent is strong HC1 containing 2 parts of ferric chloride to the liter 

 o.i gram to 50 c.c. of HC1. 



A more exact method is to pour off the supernatant acid urine. Wash the 

 chloroform with water, then pour off as much of the supernatant water as possible 

 and add 10 c.c. of alcohol. A clear blue fluid results. 



Urobilin. Urobilin appears in considerable quantity in urine when there is 

 much destruction of red cells, as in pernicious anaemia, internal haemorrhage, and in 

 malaria cachexia. The best test is that of Schlesinger. To the unfiltered urine 

 add an equal amount of a saturated solution of zinc acetate in absolute alcohol. 

 Shake, add a few drops of Lugol's solution and filter. Fluorescence in the nitrate 

 shows the presence of urobilin. The degree of blood destruction is indicated by the 

 intensity of the fluorescence. 



Bile Pigments. A satisfactory test is that of Rosin (Trousseau). Overlay 10 

 c.c. urine with about 5 c.c. of dilute tincture of iodine (i to 10 of 95% alcohol). 

 An emerald green ring at the point of contact shows the presence of bile coloring 

 matter. 



Phenolsulphonephthalein Test for Renal Efficiency. 



Geraghty has recently stated that in 35 cases where an autopsy made it possible 

 to verify the accuracy of this test that the lesions as revealed at autopsy corre- 



