The method is as follows:--Weigh up 1 gram of the substance and dissolve in a suitable acid; dilute; and add permanganate of potash solution until tinted. Boil for some time and dilute to 100 c.c. Take a couple of Nessler tubes, holding over 100 c.c., but marked at 50 c.c.; label them "1" and "2"; and into each put 10 c.c. of the potassic sulphocyanate solution and 2 c.c. of dilute hydrochloric acid. The solutions should be colourless. To "1" add 10 c.c. of the assay solution, and dilute to the 50 c.c. mark. To the other add water, but only to within 5 or 10 c.c. of this mark. Now run in the standard ferric chloride solution from a small burette, 1 c.c. at a time, stirring after each addition till the colour is nearly equal to that of the assay (No. 1). At this stage bring the solution to the same level by diluting, and make a further addition of the standard ferric chloride solution till the colours correspond. The amount of iron will be the same in each tube; that in the standard may be known by reading off the volume from the burette and multiplying by 0.01 milligram. If the 10 c.c. of the assay solution gave a colour requiring more than 5 or 6 c.c. of the standard ferric chloride solution, repeat the determination, taking a smaller proportion. The effect of varying conditions on the assay will be seen from the following experiments:-- ~Effect of Varying Temperature.~--The effect of increase of temperature is to lessen the colour; in fact, by boiling, the colour can be entirely removed. All assays are best carried out in the cold. 1 c.c. at 15° would only show the colour of 0.75 c.c. at 45° 2 " " " " 1.75 " 5 " " " " 4.0 " ~Effect of Time.~--The effect of increase of time is to increase the colour, as will be seen from the following experiments:-- 2 c.c. on standing 10 minutes became equal to 2.25 c.c. 2 " 20 " " " 2.75 " 2 " 40 " " " 3.00 " ~Effect of Free Acid.~--If no acid at all be present, the sulphocyanate of potassium solution removes the colour it first produces, so that a certain amount of acid is necessary to develop the colour. The use of a large excess has a tendency to increase the colour produced. 5 c.c. nitric acid (sp. g. 1.4) read 3.7 c.c. instead of 2 c.c. with the dilute acid. 5 c.c. sulphuric acid (sp. g. 1.32) read 2.2 c.c. instead of 2 c.c. with the dilute acid. 5 c.c. hydrochloric acid (sp. g. 1.16) read 2.5 c.c. instead of 2 c.c. with the dilute acid.