100 ppm std =

POREWATER SULFIDE ANALYSIS PROTOCOL 2.5 mL 1000 ppm +

Updated 7-28-2016 by S. Widney 22.5 mL water +
25 mL SAOB
Sample Collection in the Field:
1. Dilute samples 1:1 with sulfide anti-oxidant buffer (SAOB) (mixed with ascorbic acid) as they are
collected, to prevent the oxidation of sulfide. Add 20 mL SAOB to a 50ml centrifuge tube, then add 20
mL of sample to prepared tube.

Reagents Needed for Lab Analysis:

1. Milli-Q water in a squeeze bottle
2. Sodium sulfide (stored in fridge)
3. SAOB (one 475 mL bottle)
4. 0.4061 g lead perchlorate (to make 10 mL 0.1 M lead perchlorate)
5. Optimum B solution for electrode
6. Will also need clean beakers (at least 5) and graduated cylinders, pipette tips, disposable glass pipettes
Standard Solutions:
1. (Instruction Manual, p.5) Prepare a stock solution of saturated sodium sulfide by dissolving 10 g Na2S .
9H2O in 5 mL distilled, deaerated water and 5 mL SAOB (do in anaerobic chamber if possible). Measure
Stock Na2S . 9H2O with a non-metal spatula. Stir well and let stand overnight. Store in a tightly stoppered
solution bottle in the hood or the refrigerator for longterm storage.
a. Na2S is stored in the fridge. Use an Erlenmeyer flask (not volumetric) to avoid spillage.
2. Prepare Orion Silver/Sulfide Combination Electrode according to instructions on p.6-8 in Instruction
Manual. Fill electrode with Optimum B filling solution. Connect electrode to pH meter and adjust the
Electrode meter to measure mV. Before measuring, check the slope of the electrode according to instructions on
prep page 9 in Instruction Manual (will need to prepare 100 ppm standard for this step).
3. Prepare a sulfide standard weekly by pipetting 1 ml of the stock solution into a 100 mL volumetric flask.
Add 50ml SAOB and dilute to volume with Milli-Q.
a. Determine the exact concentration by titrating 10ml of standard with 0.1 M lead perchlorate.
Weekly std
i. To make 0.1 M lead perchlorate, dissolve 0.4061g lead perchlorate in 10ml distilled H20.
ii. Pipette 10ml of the weekly standard into a small beaker and insert the electrode. Add
lead perchlorate solution in 0.25ml increments (using automatic pipetter). Record the
reading in mV after each addition. Reading will drop from around –920mV to around
–730mV at some point. Graph these data in Excel to see the drop.

1000 Use the following equation to determine the concentration of

950 the standard:
900
850 C = 3206(Vt/Vs)
mV (-)

800
750
where:
700 C = concentration (ppm sulfide)
650 Vt = volume of titrant at end
600 point (ie. ml of lead perchlorate
0.75

1.25

1.75

3.25
0.25

2.25

2.75
0.5

2.5
1.5

3.5
0

added at the point where the

m l titrant (lead perchlorate) slope of the curve is greatest.
Use the average of the values on
either side of the drop: (2.25+2.5)/2 = 2.375)
Vs = volume of standard (10ml)
In this case, C = 3206(2.375/10) = 761.4 ppm

4. by serial dilution of the weekly standard (~1000ppm, specific ppm will vary depending on initial
concentration of weekly standard, determined with the equation above). See Standard Preparation
section below for detailed instructions.
1
 Need 25 mL graduated cylinder for
SAOB & water and one for 1000 ppm
Standard Preparation (serial dilution):
std
1000 ppm = 25 mL 1000 ppm stock (weekly standard) + 25 mL SAOB
5 beakers (at least 100 mL)
100 ppm = 2.5 mL 1000 ppm +22.5 mL water + 25 mL SAOB
5 mL pipette and tips
10 ppm = 2.5 mL 100 ppm + 22.5 mL water + 25 mL SAOB
Kimwipes
1 ppm = 2.5 mL 10 ppm +22.5 mL water + 25 mL SAOB
0.1 ppm = 2.5 mL 1 ppm + 22.5 mL water +25 mL SAOB
Prepare standards in beakers. Add 25 mL SAOB to all standards AFTER the serial dilution has been completed.

Sulfide Measurement:
1. All standards and samples should be at the same temperature for precise measurement. Remove samples
from freezer and allow them to reach room temperature before measuring sulfide.
Standards:
2. Rinse electrode with distilled water, blot dry and place into the beaker containing the most dilute
standard. When a stable reading is displayed, record the mV value and corresponding standard
concentration.
3. Repeat step 2 for all standards in order of increasing concentration (ppm). Create a calibration curve by
graphing the mV values against concentration in Excel and add a logarithmic trendline to get the
equation for the line (see below). Positive mV values may be used for easier computation without
changing the resulting concentrations (this would give the equation y = 12.959Ln(x) + 821.89).
Solve the equation for x: x = exp((y-821.89)/12.959).

-920
-900
-880
-860
mV

-840
-820
-800
-780
0.01 0.1 1 10 100 1000
ppm sulfide y = -12.959Ln(x) - 821.89
2
R = 0.9996

Samples:
4. Rinse electrode with distilled water, blot dry and place into the 50ml centrifuge tube containing the first
sample. Do not add more SAOB before measuring, since samples have already been preserved with
SAOB. When a stable reading is displayed, record the mV value. Use the equation for x obtained in
step 4 to determine the concentration of each sample. Rinse electrode with distilled water between each
sample. After measuring all samples, re-measure a few for QC.

Waste Disposal: All sodium sulfide and lead perchlorate liquid waste should be disposed of in waste bottles
in the hood, and collected by IU Environmental Health and Safety. See Sulfide Analysis Template for
hazardous waste components (update as necessary if components change significantly). Put paper towels, etc in
a container to prevent fumes

2
Storage of Orion Silver/Sulfide Combination Electrode:
The Optimum B solution in the Orion Model 9616 Combination Silver/Sulfide Electrode should not be allowed
to evaporate, causing crystallization.

For short periods of time (up to one week): Store the electrode in distilled water.

For storage longer than one week: Drain the electrode; flush the inside with distilled water and store dry with
the cap on to protect the sensing element. To drain the electrode, hold by the cap and push the plastic cap down
onto the glass tube (filling solution will drain out of the bottom). Flush several times with distilled water, then
let dry before putting away.

Notes: Standard preparation, measurement of standard curve, and measurement of samples can be done all in
the same day (will likely take most of the day). Make the stock solution the day before, and proceed with Step 2
the next day. Make sure you set samples out to warm up the night before you want to measure them.
 Preparing the stock takes ~1 hour
 Weekly standard preparation and titration takes ~2 hours
 Serial dilution of standards and standard measurement takes ~1.5 hours
 Measuring 31 samples takes ~2-3 hours

Methods adapted from Orion Model 9416 Silver/Sulfide Half-Cell and Model 9616 Sure-Flow Combination
Silver/Sulfide Electrodes Instruction Manual pp 5, 8, 39-40 (Copyright 1997, Orion Research, Inc.)