SULFIDE (4500-S2⫺)/Methylene Blue Method

color. Sulfide itself prevents the reaction if its concentration is 3. Procedure

very high, in the range of several hundred milligrams per liter.
To avoid the possibility of false negative results, use the a. Put 0.20 mL (4 drops) zinc acetate solution and 0.10 mL
antimony method to obtain a qualitative result in industrial (2 drops) 6N NaOH into a 100-mL glass bottle, fill with sample,
wastes likely to contain sulfide but showing no color by the and add 0.10 mL (2 drops) 6N NaOH solution. Stopper with no
methylene blue method. Iodide, which is likely to be present air bubbles under stopper and mix by rotating back and forth
in oil-field wastewaters, may diminish color formation if its vigorously about a transverse axis. For the iodometric procedure,
concentration exceeds 2 mg/L. Many metals (e.g., Hg, Cd, use a 500-mL bottle or other convenient size, with proportionally
Cu) form insoluble sulfides and give low recoveries. larger volumes of reagents. Vary volume of reagents added
Eliminate interferences due to sulfite, thiosulfate, iodide, and according to sample so the resulting precipitate is not exces-
many other soluble substances, but not ferrocyanide, by first pre- sively bulky and settles readily. Add enough NaOH to raise the
cipitating ZnS, removing the supernatant, and replacing it with pH above 9. Let precipitate settle for 30 min. The treated sample
distilled water. Use the same procedure, even when not needed for is relatively stable and can be held for several hours. However,
removal of interferences, to concentrate sulfide. The automated if much iron is present, oxidation may be fairly rapid.
methylene blue method (4500-S2⫺.E) is relatively free from inter- b. If the iodometric method is to be used, collect precipitate on a
ferences because gas dialysis separates the sulfide from the sample glass fiber filter and continue at once with titration according to the
matrix. procedure of 4500-S2⫺.F. If the methylene blue method
(4500-S2⫺.D) is used, let precipitate settle for 30 min and decant as
much supernatant as possible without loss of precipitate. Refill
1. Apparatus bottle with distilled water, shake to resuspend precipitate, and
quickly withdraw a sample. If interfering substances are present in
Glass bottles with stoppers: See 4500-S2⫺.B.1. high concentration, settle, decant, and refill a second time. If sulfide
concentration is known to be low, add only enough water to bring
volume to one-half or one-fifth of original volume. Use this tech-
2. Reagents nique for analyzing samples of very low sulfide concentrations.
After determining the sulfide concentration colorimetrically, multi-
a. Zinc acetate solution: Dissolve 220 g Zn(C2H3O2)2 䡠 2H2O ply the result by the ratio of final to initial volume. No concentration
in 870 mL water; this makes 1 L solution. or pretreatment steps to remove interferences are necessary for
b. Sodium hydroxide solution (NaOH), 6N. 4500-S2⫺.E.

4500-S2⫺ D. Methylene Blue Method

1. Apparatus supply may be oxidized and discolored to a degree that results in

interfering colors in the test. Store in a dark glass bottle. When
a. Matched test tubes, approximately 125 mm long and 15 mm this stock solution is diluted and used in the procedure with a
OD. sulfide-free sample, it first will be pink but then should become
b. Droppers, delivering 20 drops/mL methylene blue solution. colorless within 3 min.
To obtain uniform drops hold dropper in a vertical position and b. Amine-sulfuric acid reagent: Dilute 25 mL amine-sulfuric
let drops form slowly. acid stock solution with 975 mL 1 ⫹ 1 H2SO4. Store in a dark
c. If photometric rather than visual color determination will be glass bottle.
used, either: c. Ferric chloride solution: Dissolve 100 g FeCl3 䡠 6H2O in
1) Spectrophotometer, for use at a wavelength of 664 nm with 40 mL water.
cells providing light paths of 1 cm and 1 mm, or other path d. Sulfuric acid solution (H2SO4), 1 ⫹ 1.
lengths, or e. Diammonium hydrogen phosphate solution: Dissolve 400 g
2) Filter photometer, with a filter providing maximum trans- (NH4)2HPO4 in 800 mL distilled water.
mittance near 660 nm. f. Methylene blue solution I: Use USP grade dye or one
certified by the Biological Stain Commission. The dye content
2. Reagents should be reported on the label and should be 84% or more.
Dissolve 1.0 g in distilled water and make up to 1 L. This
a. Amine-sulfuric acid stock solution: Dissolve 27 g solution will be approximately the correct strength, but because
N,N-dimethyl-p-phenylenediamine oxalate* in an iced mixture of variation between different lots of dye, standardize against
of 50 mL conc H2SO4 and 20 mL distilled water. Cool and dilute sulfide solutions of known strength and adjust its concentration
to 100 mL with distilled water. Use fresh oxalate because an old so 0.05 mL (1 drop) ⫽ 1.0 mg sulfide/L.
Standardization—Prepare five known-concentration sulfide
standards ranging from 1 to 8 mg/L as described in
* Eastman Cat. No. 5672 has been found satisfactory for this purpose. 4500-S2⫺.A.6, or proceed as follows: Put several grams of clean,

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 SULFIDE (4500-S2⫺)/Gas Dialysis, Automated Methylene Blue Method

washed crystals of Na2S 䡠 9H2O into a small beaker. Add some- dropwise, to the second tube, until color matches that developed
what less than enough water to cover crystals. Stir occasionally for in first tube. If the concentration exceeds 20 mg/L, repeat test
a few minutes, then pour solution into another vessel. This solution with a portion of sample diluted tenfold.
reacts slowly with oxygen but the change is insignificant if analysis With methylene blue solution I, adjusted so 0.05 mL (1 drop)
is performed within a few hours. Prepare solution daily. To 1 L ⫽ 1.0 mg S2⫺/L when 7.5 mL of sample are used:
distilled water, add 1 drop of Na2S solution and mix. Immediately
determine sulfide concentration by the meth-ylene blue procedure mg S2⫺/L ⫽ no. drops solution I ⫹ 0.1 (no. drops solution II)
and by the iodometric procedure. Repeat, using more than
1 drop Na2S solution or smaller volumes of water, until at 2) Photometric color measurement—A cell with a light path of
least five tests have been made, with a range of sulfide 1 cm is suitable for measuring sulfide concentrations from 0.1 to
concentrations between 1 and 8 mg/L. Calculate average 2.0 mg/L. Use shorter or longer light paths for higher or lower
percent error of the methylene blue result as compared to the concentrations. This method is suitable for sample concentra-
iodometric result. If the average error is negative (i.e., methylene tions up to 20 mg/L. Zero instrument with a portion of treated
blue results are lower than iodometric results), dilute methylene sample from Tube B. Prepare calibration curves on basis of
blue solution by the same percentage, so a greater volume will be colorimetric tests made on Na2S solutions simultaneously ana-
used in matching colors. If methylene blue results are high, increase lyzed by the iodometric method, plotting concentration vs.
solution strength by adding more dye. absorbance. A linear relationship between concentration and
g. Methylene blue solution II: Dilute 10.00 mL of adjusted absorbance can be assumed from 0 to 1.0 mg/L.
methylene blue solution I to 100 mL with reagent water. Read sulfide concentration from calibration curve.

3. Procedure 4. Precision and Bias

a. Color development: Transfer 7.5 mL sample to each of two In a study by two chemists working in the same laboratory, the
matched test tubes, using a special wide-tip pipet or filling to marks standard deviation estimated from 34 sets of duplicate sulfide
on test tubes. If sample has been preserved with zinc acetate, shake measurements was 0.04 mg/L for concentrations between 0.2
vigorously before taking subsample. Add to Tube A 0.5 mL and 1.5 mg/L. The average recoveries of known additions were
amine-sulfuric acid reagent and 0.15 mL (3 drops) FeCl3 92% for 40 samples containing 0.5 to 1.5 mg/L and 89% for
solution. Mix immediately by inverting slowly, only once. (Excessive samples containing less than 0.1 mg/L.
mixing causes low results by loss of H2S as a gas before it has had
time to react). To Tube B add 0.5 mL 1 ⫹ 1 H2SO4 and 0.15 mL 5. Quality Control
(3 drops) FeCl3 solution and mix. The presence of S2⫺ will be
indicated by the appearance of blue color in Tube A. Color devel- The quality control practices considered to be an integral part
opment usually is complete in about 1 min, but a longer time often of each method are summarized in Table 4020:I.
is required for fading out of the initial pink color. Wait 3 to 5 min
and add 1.6 mL (NH4)2HPO4 solution to each tube. Wait 3 to 15
min and make color comparisons. If zinc acetate was used, wait 6. Bibliography
at least 10 min before making a visual color comparison. POMEROY, R.D. 1936. The determination of sulfides in sewage. Sewage
b. Color determination: Works J. 8:572.
1) Visual color estimation—Add methylene blue solution I or NUSBAUM, I. 1965. Determining sulfides in water and waste water. Water
II, depending on sulfide concentration and desired accuracy, Sewage Works 112:113.

4500-S2⫺ E. Gas Dialysis, Automated Methylene Blue Method

1. Apparatus b. N,N-dimethyl-p-phenylenediamine working solution: Dilute

190 mL N,N-dimethyl-p-phenylenediamine stock solution to 1 L.
Automated analytical equipment: An example of the continu- Store in an amber bottle. Prepare weekly.
ous-flow analytical instrument consists of the interchangeable c. Ferric chloride stock solution: Dissolve 13.5 g
components shown in Figure 4500-S2⫺:2. FeCl3 䡠 6H2O in 500 mL 5N HCl. Store in an amber bottle.
The sampler is equipped with a mixer to stir samples before Prepare fresh monthly.
analysis and the gas dialysis membrane, which is maintained at d. Working ferric chloride solution: Dilute 190 mL ferric
room temperature, separates H2S from the sample matrix. chloride stock solution to 1 L. Store in an amber bottle. Prepare
fresh weekly.
2. Reagents
e. Hydrochloric acid (HCl), 6N.
a. N,N-dimethyl-p-phenylenediamine stock solution: Dissolve f. Sodium hydroxide stock solution (NaOH), 1N.
1 g N,N-dimethyl-p-phenylenediamine dihydrochloride in 500 mL g. Sodium hydroxide (NaOH), 0.01N: Dilute 10 mL NaOH
6N HCl. Prepare fresh monthly. Store in an amber bottle. stock solution to 1 L.

https://doi.org/10.2105/SMWW.2882.096 5