15preferably performed with a pipetteto minimize mixing with oxygen, anda maximum of 10% of the vial volumeshould be left as headspace Close thecontainer with a gas tight lid and shakevigorously3 Obtain zero readingThe signal at zero H2S can be obtained by immersing the sensor tipinto one of the vials with calibration buffer Note the signal, whichis the calibration value for zero H2S partial pressure (S0) This signalshould be 0-30 pA (otherwise see “Troubleshooting”)4 Obtain sulfide standard readingsCalibration points within the expected range of measurement areprepared by injecting suitable amounts of the S2- stock solutionanaerobically into the calibration buffer with a micro-syringe(the stock solution should be diluted at least 10 times) Mix thesolutions If you have not added reductant, oxygen that dissolvein the calibration solution will oxidize the sulfide, so the stocksolution should be added immediately after the oxygen removal(e g N2 bubbling) and the calibration should be done immediatelyafter adding the stock solutionFor each calibration solution, measure the calibration valuesby removing the rubber stopper and immerse the microsensortip into the solution Read the signal and plot it against theconcentrationThe most precise calibration curve is obtained by fixingthe calibration solutions with Zn-acetate and subsequentlydetermining the total sulfide concentration using the Clinemethod (Cline 1969) This may, due to the nature of the method,be difficult if a reductant has been added previouslycalibration in the fieldFor an easy and portable field calibration, prepare the followingreagents in the lab Bring the reagents and a 1 ml syringe + needleto the field