Hydrogen Sulfide (Zinc Acetate Procedure)

1. Prepare a 1:20 zinc acetate working solution in distilled water - 7 ml of 22% stock
solution (Fisher) to 140 mL of DI water for working solution).
2. Prepare the diamine reagent, storing it in a dark bottle in the refrigerator:
a. 4 gram FeCl3
b. 1.6grams N,N-dimethyl phenylene diamine sulfate
c. 100 mL 6N HCl
3. Label test tubes with each sample ID. Add sample aliquot up to 1 mL total
(depending on the anticipated sulfide concentration), making up the volume to 1
mL with distilled water. e.g. 0.5 mL sample and 0.5 mL of DI water.
4. Prepare 2 blank samples for zeroing of the spectrophotometer. Add 1 mL of
distilled water to each blank.
5. Add 5 mL of 1:20 Zn acetate solution to each sample.
6. Add 0.4 mL of the diamine reagent to each sample.
7. Incubate samples and blank solutions for 20 minutes for color development.
8. Turn on spectrophotometer so it can warm up for at least 10 minutes prior to use.
Set the spectrophotometer at 670 nm.
9. Fill the disposable spectrophotometer tubes with the blanks and place into the
spec. Zero the spectrophotometer with the blanks by hitting “AUTOZERO”.
10. Leave one blank in spectrophotometer while running samples. Run the samples
and record the absorbance.
11. The regression equation for the spec in Lab 408 is as follows:
Ppm sulfide = ((12.244 x absorbance) – 1.4523) x 2