458 APPENDIX 



t in the 



10 dilution of the urine, mix and draw up with a bulb pipette and deposit i 

 short arm of the U tube containing nitric acid. A distinct ring forms in two to 

 three seconds. Pour off one-half of the diluted urine and make up with an equal 

 amount of saline. Deposit this i to 20 dilution in the long arm. The ring forms 

 in about a minute. With further testing it is found that a i to 40 dilution shows a 

 perceptible ring in just two minutes. This final and successful dilution multiplied 

 by 0.0033 gives the percentage of albumin in the urine (40 X 0.0033 = 0.13%). 



Should it be desired to determine the nature of the proteids present either in 

 urine or in exudates or transudates the following method is applicable. Determine 

 the percentage of total proteid by the method employed above. Then throw down 

 the globulins by the addition of an equal amount of a saturated solution of ammo- 

 nium sulphate, filter and estimate the proteid content of the filtrate. The differ- 

 ence between that and the total gives the percentage of globulin. The filtrate is 

 now treated with 5% acetic acid until a precipitate of nucleo-proteid ceases to form; 

 the fluid is filtered and the clear filtrate (which should not show any turbidity with 

 a drop of 5% acetic acid) is tested for its proteid content, which represents the 

 serum albumin. When the combined percentage of globulins and serum albumin 

 is subtracted from the total proteid percentage we have the percentage of nucleo- 

 proteid. 



Esbach's Quantitative Method for Albumin. The use of the Esbach tube is at- 

 tended with some uncertainty, whether using the original Esbach solution or that 

 devised by Tsuchiya, the precipitate at times refusing to settle. The method of 

 using the tube is to add the urine to the U mark and then the reagent to the R mark, 

 mix, and allow to stand in upright position. If Esbach's reagent has been used the 

 reading is made at the end of twenty-four hours, but when Tsuchiya's reagent is 

 employed the reading is made at the end of two hours. The number at the end of 

 the line which corresponds to the upper limit of precipitate will be the number 

 of grams of dry albumin per liter of urine. Esbach's reagent consists of 10 grams 

 of picric and 20 grams of citric acid dissolved in i liter of water. Tsuchiyas' 

 reagent is made by dissolving 1.5 grams of phosphotungstic acid in a mixture of 5 

 c.c. strong hydrochloric acid and 95 c.c. of 95% alcohol. In this method also 

 dilution must be resorted to if albumin or specific gravity are excessive. 



Nucleo -protein. Increased quantities of this protein occur in pyelitis, nephritis, 

 and inflammations of the bladder. 



For its detection albumin, if present in any considerable quantity, must be re- 

 moved by boiling as described above. Then place 10 c.c. of this urine in a small 

 beaker, dilute with 3 volumes of water and make the reaction very strongly 

 acid with acetic acid. If the solution becomes turbid it is an indication that 

 nucleo-protein is present. 



Bence-Jones Body. (Albumose.) Perform the heat test for albumin. The 

 appearance of a heavy precipitate which partially clears on boiling suggests albu- 

 mose. If albumose is present a cloud will appear in the filtrate on cooling. The 

 precipitate formed with nitric acid, if due to albumose, disappears with heat, that 

 of serum albumin does not. 



As another test for the Bence-Jones body, usually present in multiple myelomata, 

 that of Boston is of value. Mix 15 c.c. urine in a test-tube with an equal amount of 

 saturated NaCl solution. Add 2 c.c. of 40% NaOH solution and shake the contents 



