Instruction Manual Calcium Electrode EUTECH INSTRUMENTS PTE LTD. INSTRUCTION MANUAL CALCIUM ION ELECTRODE 1 Calcium Electrode Instruction Manual TABLE OF CONTENTS GENERAL INSTRUCTIONS .......................................................................................................... 3 Introduction ..............................................................................................................................................3 Required Equipment.................................................................................................................................3 Required Solutions ...................................................................................................................................3 GENERAL PREPARATION............................................................................................................ 4 Electrode Preparation ...............................................................................................................................4 Electrode Slope Check (with pH/mV meter) ...........................................................................................4 Electrode Slope Check (with ion meter) ..................................................................................................5 MEASUREMENT.............................................................................................................................. 5 Measuring Hints .......................................................................................................................................5 Sample Requirements...............................................................................................................................6 Units of Measurement ..............................................................................................................................6 MEASUREMENT PROCEDURE ................................................................................................... 6 Direct Measurement .................................................................................................................................6 Direct Measurement of Calcium (using a pH/mV meter) ........................................................................6 Direct Measurement of Calcium (using an ion meter) .............................................................................8 Low Level Calcium Determination (using a pH/mV meter)....................................................................8 Low Level Calcium Determination (using an ion meter).........................................................................9 Titration....................................................................................................................................................9 Titration of Calcium ...............................................................................................................................10 ELECTRODE CHARACTERISTICS........................................................................................... 11 Reproducibility.......................................................................................................................................11 Interferences ...........................................................................................................................................11 Complexation .........................................................................................................................................12 Temperature Influences..........................................................................................................................12 Electrode Response ................................................................................................................................13 Limits of Detection.................................................................................................................................13 Electrode Life.........................................................................................................................................13 Electrode Storage ...................................................................................................................................14 ELECTRODE THEORY ................................................................................................................ 14 Electrode Operation ...............................................................................................................................14 TROUBLESHOOTING GUIDE .................................................................................................... 15 Meter ......................................................................................................................................................15 Glass-ware..............................................................................................................................................15 Electrodes...............................................................................................................................................15 Standards and Reagents..........................................................................................................................16 Sample....................................................................................................................................................16 Technique...............................................................................................................................................16 TROUBLESHOOTING HINTS..................................................................................................... 17 SPECIFICATIONS.......................................................................................................................... 19 ORDERING INFORMATION....................................................................................................... 19 2 Instruction Manual Calcium Electrode EUTECH INSTRUMENTS PTE LTD. CALCIUM ELECTRODE INSTRUCTION MANUAL GENERAL INSTRUCTIONS Introduction Eutech Instruments Calcium Ion Electrode is used to measure calcium ions in aqueous solutions quickly, simply, accurately, and economically. Required Equipment 1. 2. A pH/mV meter or an ion meter, either line operated or portable. Semi-logarithmic 4-cycle graph paper for preparing calibration curves when using the meter in the mV mode. A magnetic stirrer. Eutech Calcium Epoxy-body Combination Electrode, Code. no. EC-CAL-03. 3. 4. Required Solutions 1. 2. Deionized or distilled water for solution and standard preparation. Eutech Calcium Standard, 0.1M CaCl2, Code. no. EC-SCS-CA1-BT. To prepare this solution from your own laboratory stock, half fill a one liter volumetric flask with distilled water and add 14.7 grams of reagent-grade calcium chloride (CaCl2.2H2O). Swirl the flask gently to dissolve the solid. Fill to the mark with distilled water, cap, and upend several times to mix the solution. Eutech Calcium Standard, 1,000 ppm Ca+2, Code. no. EC-SCS-CA2-BT. To prepare this solution from you own laboratory stock, half fill a one liter volumetric flask with distilled water and add 3.67 grams of reagent-grade calcium chloride (CaCl2.2H2O). Swirl the flask gently to dissolve the solid. Fill to the mark with distilled water, cap, and upend several times to mix the solution. Eutech Calcium Standard, 100 ppm Ca+2 as CaCO3, Code. no. EC-SCS-CA3-BT. To prepare this solution from your own laboratory stock, half fill a one liter volumetric flask with distilled water and add 0.15 grams of reagent-grade calcium chloride (CaCl2.2H2O). Swirl the flask gently to dissolve the solid. Fill to the mark with distilled water, cap, and upend several times to mix the solution. Eutech Ionic Strength Adjuster (ISA), 4 M KCl, Code. no. EC-ISA-CA1-BT. To prepare this solution from your own laboratory stock, half fill a 1,000 ml volumetric flask with distilled water and add 298 grams of reagent-grade potassium chloride (KCl). Swirl the 3. 4. 5. 3 Calcium Electrode Instruction Manual flask gently to dissolve the solid. Fill to the mark with distilled water, cap, and upend several times to mix the solution. 6. EDTA titrant, 1M stock solution, for the titration of calcium. To prepare this titrant, add 37.2 grams of reagent grade Na2EDTA.2H2O, ethylenediaminetetraacetic acid dihydrate, disodium salt, to a 100 ml volumetric flask, add about 75 ml of distilled water, and swirl the flask gently to dissolve the solid. Fill to the mark with distilled water, cap, and upend several times to mix the solution. GENERAL PREPARATION Electrode Preparation Remove the rubber caps covering the electrode tips and the rubber insert covering the filling hole of the reference electrode. Fill the combination electrode or the reference electrode with the filling solution shipped with the electrode to a level just below the fill hole. No preparation is required with a sealed reference electrode. Gently shake the electrode downward in the same manner as a clinical thermometer to remove any air bubbles which might be trapped behind the calcium membrane. Prior to first usage, or after long-term storage, immerse the calcium membrane in calcium standard for thirty minutes. The electrode is now ready for use. Connect the electrodes to the proper terminals as recommended by the meter manufacturer. Electrode Slope Check (with pH/mV meter) (Check electrodes each day) 1. To a 150 ml beaker, add 100 ml of distilled water. Place the beaker on a magnetic stirrer and begin stirring at a constant rate. After assuring that the meter is in the millivolt mode, lower the electrode tips into the solution. If drifting or instability is observed, see the TROUBLESHOOTING section. Using a pipet, add 1 ml of 0.1M, 1,000 ppm, or 100 ppm (as calcium carbonate) standard and 2 ml of ISA to the beaker. When the reading is stable, record the mV reading. Using a pipet, add 10 ml of the same calcium standard used above to the beaker. When the reading has stabilized, record the mV reading. Determine the difference between the two readings. The electrode is operating correctly if the mV potential has changed by 27±2 mV, assuming the solution temperature is between 20o and 25oC. See the TROUBLESHOOTING section if the potential change is not within this range. 2. 3. 4. Slope is defined as the change in potential observed when the concentration changes by a factor of 10. 4 Instruction Manual Calcium Electrode Electrode Slope Check (with ion meter) (Check electrodes each day) 1. Prepare standard calcium solutions whose concentrations vary by tenfold. Use either the 0.1M Ca+2, 1,000 ppm Ca+2, or the 100 ppm Ca+2 (as calcium carbonate) standard stock solutions. Use the serial dilution method for this preparation. To a 150 ml beaker, add 100 ml of the lower value standard and 2 ml of ISA. Place the beaker on the magnetic stirrer and begin stirring at a constant rate. Lower the electrode tips into the solution. Assure that the meter is in the concentration mode. Adjust the meter to the concentration of the standard and fix the value in the memory according to the meter manufacturer's instructions. Rinse the electrodes with distilled water and blot dry. To another 150 ml beaker, add 100 ml of the higher value standard and 2 ml of ISA. Place the beaker on the magnetic stirrer and begin stirring at a constant rate. Lower the electrode tips into the solution. Adjust the meter to the concentration of the standard and fix the value in the memory. Read the electrode slope according to the meter manufacturer's instructions. Correct electrode operation is indicated by a slope of 90-100%. See the TROUBLESHOOTING sections if the slope is not within this range. 2. 3. 4. 5. 6. 7. MEASUREMENT Measuring Hints All samples and standards should be at the same temperature for precise measurement. A difference of 1oC in temperature will result in a 4% measurement error. The sensing membrane is normally subject to water uptake and might appear milky. This has no effect on performance. Constant, but not violent, stirring is necessary for accurate measurement. Magnetic stirrers can generate sufficient heat to change the solution temperature. To counteract this effect, place a piece of insulating material, such as a styrofoam sheet, between the stirrer and beaker. Always rinse the electrode tips with distilled water and blot dry. Use a clean, dry tissue to prevent cross-contamination. For samples with high ionic strength, prepare standards with compositions similar to that of the sample. Always check to see that the membrane is free from air bubbles after immersion into standard or sample. A slow responding electrode may be caused by interferences to the electrode. To restore proper performance, soak the electrode in distilled water for about 5 minu ...