Application Guide: Titration Automation in the Food and Beverage Industry
AG C Bo ok 2 0% C ya n 45 Application Brochure Titration Excellence Workflow Automation Multi-Parameter Analyses Selected Applications Fully Automated – Secure and Efficient Fo od a nd B ev er ag es METTLER TOLEDO METTLER TOLEDO I Automation in Food & Beverages Introduction Around the world, the Food and Beverages Industry continues to expand as the food need constantly increases as a result of the combined effects of world population growth, rising incomes and dietary habit changes. At the same time there is an increasing demand for high and consistent quality, appearance and taste of the different food products. This puts pressure on the food industry to find as secure and efficient measurement solutions as possible. Food and Beverages Industry is subject to regulations covering the whole production process for the benefit of food safety, consumer protections and (inter)national trade. The strict quality standards in the Food and Beverages Industry require running several quality control tests on the products before they reach the consumer. For this reason, well-defined analytical reference methods are needed at all stages of the production chain. Among other techniques, content determination by titration is a widely used analytical technique, in particular for routine analysis of raw materials as well as final products. Titration is a quantitative analytical method. The main advantages of titrimetric methods such as the linearity over the whole measuring range and the high precision are well appreciated benefits for the user. Equally important for the user are the many titration methods available for the different Food and Beverages samples which are tested and approved by a long history of practice (see also Application Brochure 31 with a large collection of titration methods for the Food and Beverages Industry). Titration comes nowadays with a large extent of automation from sample preparation to addition of reagents, result calculations and data storage. METTLER TOLEDO's LabX® software guarantees data integrity and fully supports you with highest regulations (including 21 CFR part 11). Usage of the SmartSampleTM solution will further increase process security: at the balance all the sample data are written onto an RFID-chip (the SmartTagTM) attached to the beaker. The InMotionTM Autosampler, equipped with an RFID reader, automatically extracts this information and transfers it to the titrator. This secures sample and data handling before, during and after analysis. Furthermore, METTLER TOLEDO instruments are fully modular. They can be combined, making it possible to fully automate the determination of multiple parameters with just one click instead of carrying out these routine analyses on separate instruments. This saves time and work power – both important factors in the fast developing industry. We, at the Market Support Group Analytical Chemistry are proud to present to you our Application Brochure 45. You can expect detailed descriptions of selected fully automated determinations in the Food and Beverages Industry via titrimetric analysis as well as combined multi-instrument systems. Daniel Buchmann Julia Werder Head Market Support Group Applications Chemist SBU Analytical Chemistry Market Support Group METTLER TOLEDO METTLER TOLEDO - II - Automation in Food & Beverages Content Method Title Multi-parameter Analysis M677 Automated Acidity, pH and Brix analysis with RM40, T90 and InMotion Flex Two ways to get fast and reliable results for acidity, Brix and pH in one go High Throughput Automation M676 High Throughput Free and Total Sulfur Dioxide (SO2) Content in Wines Novel method to speed up the fee and total SO2 determination in wine using two InMotion Max M674 Titrant Concentration Determination by Automated Volumetric KF Titration M675 Water Content in Food Products Determination by Automated Volumetric KF Titration Automated water content determination of honey, jam and cacoa M657 Parallel Tasking: Sample Preparation and Titration with Two Towers Ketchup sample is titrated for chloride at tower B while at the same time the next sample is prepared at tower A resulting in a time saving of 45 – 50% M658 Parallel Tasking with Two Towers Combined acid is determined at tower A, while the chloride content is determined simultaneously at tower B M673 High Throughput with InMotion Max: Chloride Content Determination Robust method suitable for fully automated measurement of a large number of samples per day Ion Content using Multiple Standard Addition Technique M703 Automated Sodium Determination by Standard Addition with Internal Extraction (Homogenizer): Biscuits M709 Automated Calcium Determination by Standard Addition: Mineral Water M710 Automated Nitrate Determination by Standard Addition: Spinach M711 Automated Potassium Determination by Standard Addition: Milk METTLER TOLEDO Application M677 METTLER TOLEDO - Page 1 of 14 - Sodium in Food & Beverages Automated Acidity, pH and Brix analysis with RM50, T90 and InMotion Flex 2 Ways to get fast and reliable results of Acidity, Brix and pH in one go Beverage manufactures have to determine the sugar and acid content as well as the pH value of their finished juices and carbonated drinks. The sugar concentration is generally obtained by refractive index measurement. The sugar content is a very important parameter regarding the taste and it is also important to know as general "sugar-level" for the consumer. Titration is used to determine the acid content that is commonly expressed in %- citric acid or other organic acids present in the juices and drinks such as malic acid (apple juice) and tartaric acid (grape juice). Since the acidity of the juice strongly contributes to its taste, the correct acid content is very important to achieve. In order to obtain accurate results, samples require homogenization and/or decarbonation prior to analysis. This is a rather time consuming step. By using an InMotion Autosampler, this treatment can be automated, which leads to time savings and operator independency. Description of the Application General: The following application compares two different automated systems designed to analyse beverages such as carbonated drinks and fruit juices and determines the following numbers:  % Brix, which is equivalent to 1g of sucrose per 100g of solution  acid content in % citric acid  pH We have chosen two different setups to perform this comparison. The main difference between these two setups is the following: Setup 1: The titration takes place in the aliquot beaker located on the manual titration stand of the titrator. A defined volume of juice (sample loop attached to the TV6 valve) is transferred into the beaker with excess deion. water followed by titration. This setup is suitable for low volumes from 1 – 10 mL sample size. Setup 2: The titration takes place directly in the titration beaker located on the InMotion sample changer. A dedicated pump defines the volume levels of the sample. The exact volume can be determined with acid/base titrations of known concentrations. This setup is suitable for larger volumes from 20 – 60 mL sample size. Application M677 METTLER TOLEDO METTLER TOLEDO - Page 2 of 14 - Automation in Food & Beverages Sample preparation for carbonated beverage and fruit juices Carbonated beverages: These beverages require decarbonation prior to analysis to prevent carbonic acid and gaseous carbon dioxide from interfering with refractive index measurement, pH measurement and titration. By using the InMotion Autosampler with DryPal™ decarbonation takes place directly in the sample beaker that is placed into the rack of the sample changer: The functions “stir” and “turn on the air pump” will automatically decarbonate each sample prior to analysis. While stirring with increasing speed levels, the DryPal blows air through the sample to take up and expel the carbon dioxide, thus providing an automatic, repeatable decarbonation process. Treatment of fruit juices: A homogeneous distribution in the measuring beaker is a prerequisite for accurate and repeatable measurements. Samples are often placed on a sample changer for longer time periods prior to analysis. Some of these beverage samples will contain solid particles. As a consequence, some solid particles will have settled when the samples are measured. The stirrer on the InMotion Autosampler removes this problem by homogenizing each sample right before measurement. Picture: Measurement with multi-parameter system RM40, T90 and InMotion Autosampler using setup 1 METTLER TOLEDO Application M677 METTLER TOLEDO - Page 3 of 14 - Automation in Food & Beverages Measurement principle of setup 1: Measurement principle of setup 2 Application M677 METTLER TOLEDO METTLER TOLEDO - Page 4 of 14 - Automation in Food & Beverages Measurements, Titration: The pH is measured initially via an electrode connected to the titrator in the sample beaker. Afterwards the SPR200 pump A is activated to fill the RM 40/50 flow cell and the sample loop of the TV6. In Setup 2 the excess sample is directly transferred in to the waste. The RM measures the Brix value. Subsequently the sample loop is flushed with deionized water into the titration beaker with pump B and the titration is performed to an endpoint of pH 7 with 0.1 M sodium hydroxide. In the setup 2 pump B (level pump) is activated in order to obtain the exact volume in the titration beaker. Subsequently the titrator titrates to an endpoint of pH 7 with 0.5 M sodium hydroxide. Rinsing and cleaning of system For Setup 1 the titration beaker and the sample loop are rinsed (Pump C: Drain; Pump B: Rinse). After rinsing the next sample can be analyzed. The sample in the RM loop is drained towards the waste. For Setup 1 the loop is now filled for a defined time which makes sure that the remaining sample in the loop is washed out with the new sample. With a long filling time we can assure that only the "new" sample will be present in the sample loop and in the flow through cell of the RM. For Setup 2 the tubes, stirrer and sensor on the InMotion titration stand are rinsed by PowerShower using a SD660 membrane pump and a SP280 peristaltic pump and the next sample is processed. Synchronization of refractometer (RM) and titrator (Tx, x=70 or 90): In the following flow chart the RM-Tx synchronization for Setup 1 is shown. The synchronization for Setup 2 is similar with the exception that no TV6 is used. Conclusion: Both systems give results that are of the same order of magnitude. The main advantage of setup 1 is the smaller sample size that allows to have less juice or drinks in the titration beaker located on the InMotion sample changer. On the other hand setup 2 does not require the TV6 and is over all a simpler system to maintain. METTLER TOLEDO Application M677 METTLER TOLEDO - Page 5 of 14 - Automation in Food & Beverages Send pH Result Send Acidity Result Method Synchronization: Decarbonation and analysis of acid content, pH and Brix Method RM 003 Aux. Inst.: Valve of TV6 is switched to sampling position InMotion stirs with increasing speed for decarbonation of the beverage DryPal is turned on for completing the decarbonation DryPal is turned off and stirring is continued to keep the sample homogeneous. Output sequence to start the pH- measurement Method Titration pH is measured Start Output sequence with pH Result is sent Input sequence with pH Result is received RM flow cell and sampling loop for titration are filled with pump A 014 Aux. Inst: Valve of TV6 is switched to transfer position Output sequence to start titrator method Start 2 Start of titrator method Waiting Pump B is turned on to transfer the sample into the titration beaker Refractive Index is measured EQP titration is performed Output sequence with Acidity Result is sent Input sequence with Acidity Result is received Pump C is activated to clean the titration sample loop with deion. H 2 O 012 Aux. Inst: TV6 is switched to sampling position in order to drain the RM loop Waiting Waiting Application M677 METTLER TOLEDO METTLER TOLEDO - Page 6 of 14 - Automation in Food & Beverages Preparation and Procedures Sample handling ‐ Setup 1 - TV6: Sample loop and RM50 flow cell filling and dispensing into aliquotation beaker. ‐ Setup 2 - Level Pump: RM50 flow cell filling and levelling of the volume in the titration beaker. Sample analysis ‐ Acid base titration with NaOH as titrant. ‐ Measurement of the refractive index. Remarks ‐ For setup 1 NaOH (0.1 M) is used since the sample loop volume is about 5 mL. For setup 2 the volume is about 50 mL and, therefore 0.5 M – 1.0 M NaOH should be used. ‐ The volume of the sample loop (setup 1) and of the sample volume present after levelling can be accurately determined by acid-base titration of exactly known concentrations. It is recommended to repeat this calibration on a weekly basis. Sample Fruit Juices and Beverages: Sample sizes: 6, 50 mL Compound Citric Acid M = 192.13 g/mol, z = 3 Sucrose (% Brix) Chemicals Deion. H2O Titrant Sodium hydroxide, NaOH c(NaOH)=0.1 mol/L or 0.5 mol/L. Standard Potassium hydrogen phthalate (KHP), 80 mg. Indication DGi111-SC, DGi115-SC combined pH glass electrodes Chemistry HA + NaOH = H2O + NaA Calculation Acidity: R = Q*C/m, in % C=M/(10*z) % Brix: g sucrose/100 g water Waste disposal Author, Version Angela de Martin, Thomas Hitz, MSG AnaChem, v.1.0 METTLER TOLEDO Application M677 METTLER TOLEDO - Page 7 of 14 - Automation in Food & Beverages Instruments ‐ Titration Excellence T90 (T70 is also applicable without conditions) ‐ Refractometer RM50 or RM40 (RM50: ± 0.014°Brix, RM40: ± 0.1°Brix) ‐ InMotion Flex Accessories ‐ SD660 membrane pump ‐ SP280 peristalitc pump ‐ SPR200 peristaltic reversible pump ‐ Dry Pal ‐ Tubes and accessories for all connections - A comprehensive list is given in the Appendix Results Date/Time ID nD Brix % (w/w) pH ACID % (w/w) Setup 1 08.05.2015 13:33 Orange 1.34742 9.74 2.68 0.201 08.05.2015 13:39 Orange 1.34743 9.74 2.68 0.199 08.05.2015 13:45 Orange 1.34744 9.75 2.68 0.195 08.05.2015 13:52 Orange 1.34743 9.74 2.68 0.199 08.05.2015 13:58 Orange 1.34744 9.75 2.68 0.201 Setup 2 08.05.2015 11:47 Orange 1.34736 9.70 2.69 0.193 08.05.2015 11:52 Orange 1.34735 9.69 2.69 0.190 08.05.2015 11:57 Orange 1.34734 9.68 2.69 0.190 08.05.2015 12:02 Orange 1.34736 9.70 2.69 0.194 08.05.2015 12:08 Orange 1.34736 9.70 2.69 0.194 Statistics Setup 1* Setup 2* Brix Acid (%) Brix Acid (%) Mean 9.74 0.201 9.70 0.192 s 0.024 0.0046 0.017 0.0018 Srel (%) 0.246 2.29 0.175 0.938 * without degasing steps Application M677 METTLER TOLEDO METTLER TOLEDO - Page 8 of 14 - Automation in Food & Beverages Date/Time ID nD Brix % (w/w) pH ACID % (w/w) Setup 1 06.05.2015 11:59 Cola 1.34849 10.43 2.61 0.062 06.05.2015 12:05 Cola 1.34849 10.43 2.61 0.060 06.05.2015 12:11 Cola 1.34850 10.44 2.61 0.059 06.05.2015 12:18 Cola 1.34848 10.42 2.61 0.060 06.05.2015 12:24 Cola 1.34850 10.44 2.61 0.061 Setup 2 06.05.2015 09:31 Cola 1.34839 10.37 2.63 0.048 06.05.2015 09:36 Cola 1.34839 10.37 2.62 0.049 06.05.2015 09:41 Cola 1.34841 10.38 2.62 0.047 06.05.2015 09:46 Cola 1.34839 10.37 2.62 0.048 06.05.2015 09:51 Cola 1.34838 10.36 2.63 0.050 Statistics Setup 1 Setup 2 Brix Acid (%) Brix Acid (%) Mean 10.43 0.060 10.37 0.060 s 0.008 0.001 0.007 0.001 Srel (%) 0.080 1.88 0.068 2.35 METTLER TOLEDO Application M677 METTLER TOLEDO - Page 9 of 14 - Automation in Food & Beverages Methods 001 Title Type General titration Compatible with T90 ID Setup 1 Title Acidity_Brix Author Administrator Date/Time 04/13/2015 Modified on 07/21/2015 Protect No SOP None 002 Sample Number of IDs 1 ID 1 Coca Entry type Fixed volume Volume 6.777 mL Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C 003 Titration stand (Autostand) Type Auto stand Titration stand Auto stand 1 004 Auxiliary instrument Control type RS-232 Name RM50 Send output sequence No Wait for input sequence Yes Max. time ∞ s Input sequence START Input sequence with result No Condition No 005 Measure (normal) [1] Sensor Type pH Sensor DG115-SC Unit pH Temperature acquisition Temperature acquisition No Stir Speed 30% Acquisition of the measured values Acquisition Equilibrium controlled dE 0.5 mV dt 1 s t(min) 3 s t(max) 30 s Mean value No Condition Condition No 006 Calculation R1 Result pH-value Result unit pH Formula R1=E[1] Constant C= 1 M M[None] Z z[none] Decimal places 2 Result limits No Record statistics Yes Extra statistical func. No Condition No 007 Auxiliary instrument Control type RS-232 Name RM50 Send output sequence Yes Output sequence pH%R1% Wait for input sequence Yes Max. time ∞ s Input sequence START2 Input sequence with result No Condition No 008 Pump Auxiliary reagent Deion.H2O Volume 60 mL Pump property 1-way Condition No 009 Titration (EQP) [1] Titrant Titrant NaOH Concentration 0.1 mol/L Sensor Type pH Sensor DG111-SC Unit pH Temperature acquisition Temperature measurement No Stir Speed 30% Predispense Mode None Wait time 0 s Control Control Normal Mode Acid/base Show parameters No Evaluation and recognition Procedure Standard Threshold 3 pH/mL Tendency None Ranges No Add. EQP criteria No Termination At Vmax 10 mL At potential Yes Potential 8.4 pH At slope No After number of recognized EQPs No Combined termination criteria No Accompanying stating Accompanying stating No 010 Calculation R2 Result Content Result unit % Formula R1=QE(8.2)*C/m Constant C= M/(10*z) M M[Hydrochloric acid] z z[Hydrochloric acid] Decimal places 3 Result limits No Record statistics Yes Extra statistical func. No Send to buffer No Condition No 011 Auxiliary instrument Control type RS-232 Name RM50 Send output sequence Yes Output sequence AC %R2%A Wait for input sequence No Condition Yes Formula R2>=0 012 Drain Drain pump DRAIN Drain volume 110 mL Pump property 1-way 013 Pump Auxiliary reagent Deion.H2O Volume 80 mL Pump property 1-way Condition No 014 Stir Speed 70% Duration 5 s Condition No 015 Stir Speed 0% Duration 0 s Condition No Titration Method Setup 1: Application M677 METTLER TOLEDO METTLER TOLEDO - Page 10 of 14 - Automation in Food & Beverages 016 Auxiliary instrument Control type RS-232 Name RM50 Send output sequence Yes Output sequence CL Wait for input sequence No Condition No 017 Drain Drain pump DRAIN Drain volume 130 mL Pump property 1-way Condition No 018 Record Summary No Results Per sample Raw results Per sample Table of meas. values No Sample data No Resource data No E - V No dE/dV - V No log dE/dV - V No d2E/dV2 - V No BETA - V No E - t No V - t No dV/dt - t No T - t No E - V & dE/dV - V No V - t & dV/dt - t No Method No Series data No Condition No 019 End of sample 001 Title Type General titration Compatible with T90 ID Setup 2 Title Acidity_Brix Author Administrator Date/Time 04/13/2015 Modified on 07/21/2015 Protect No SOP None 002 Sample Number of IDs 1 ID 1 Cola Entry type Fixed volume Volume 48.012 mL Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C 003 Titration stand (Autostand) Type Auto stand Titration stand Auto stand 1 004 Auxiliary instrument Control type RS-232 Name RM50 Send output sequence No Wait for input sequence Yes Max. time ∞ s Input sequence START Input sequence with result No Condition No 005 Measure (normal) [1] Sensor Type pH Sensor DG115-SC Unit pH Temperature acquisition Temperature acquisition No Stir Speed 30% Acquisition of the measured values Acquisition Equilibrium controlled dE 0.5 mV dt 1 s t(min) 3 s t(max) 30 s Condition Condition No Mean value No Condition Condition No 006 Calculation R1 Result pH-value Result unit pH Formula R1=E[1] Constant C= 1 M M[None] Z z[none] Decimal places 2 Result limits No Record statistics Yes Extra statistical func. No Condition No 007 Auxiliary instrument Control type RS-232 Name RM50 Send output sequence Yes Output sequence pH%R1% Wait for input sequence Yes Max. time ∞ s Input sequence START2 Input sequence with result No Condition No 008 Pump Auxiliary reagent Level Volume 50 mL Pump property 1-way Condition No 009 Titration (EQP) [1] Titrant Titrant NaOH_2 Concentration 0.5 mol/L Sensor Type pH Sensor DG115-SC Unit pH Temperature acquisition Temperature measurement No Stir Speed 30% Predispense Mode None Wait time 0 s Control Control Normal Mode Acid/base Show parameters No Evaluation and recognition Procedure Standard Threshold 3 pH/mL Tendency None Ranges No Add. EQP criteria No Termination At Vmax 10 mL At potential Yes Potential 8.4 pH At slope No After number of recognized EQPs No Combined termination criteria No Accompanying stating Accompanying stating No 010 Calculation R2 Result Content Result unit % Formula R1=QE(8.2)*C/m Constant C= M/(10*z) M M[Hydrochloric acid] z z[Hydrochloric acid] Decimal places 3 Result limits No Record statistics Yes Extra statistical func. No Send to buffer No Condition No Titration Method Setup 2: METTLER TOLEDO Application M677 METTLER TOLEDO - Page 11 of 14 - Automation in Food & Beverages 011 Auxiliary instrument Control type RS-232 Name RM50 Send output sequence Yes Output sequence AC %R2%A Wait for input sequence No Condition Yes Formula R2>=0 012 Auxiliary instrument Control type RS-232 Name RM50 Send output sequence Yes Output sequence CL Wait for input sequence No Condition No Formula No 013 Record Summary No Results Per sample Raw results Per sample Table of meas. values No Sample data No Resource data No E – V No dE/dV - V No log dE/dV - V No d2E/dV2 - V No BETA - V No E - t No V - t No dV/dt - t No T - t No E - V & dE/dV - V No V - t & dV/dt - t No Method No Series data No Condition No 019 End of sample Refractometer Methods (TV6, Level): 01 Title Type Measurement Compatible with RM50/RM40 ID Setup 1 Title AcidityBrix Author Administrator Date/Time 04/13/2015 Modified on 07/21/2015 Protect No SOP None 002 Configuration Cell mode RM Automation InMotion Flex Lid handling No External instruments None 003 Auxiliary instrument Control type USB-RS232 Name TV 6 Send output sequence Yes Output sequence GOA\013 Wait for input sequence No Condition No 004 Sample Sample ID Coca Correction Factor 1.0 Comment No 005 Stir Speed 1 50% Stir duration 6s Continue stirring Yes Speed 2 35% Condition No 006 Stir Speed 1 95% Stir duration 15s Continue stirring Yes Speed 2 25% Condition No 007 Auxiliary instrument Control type Output 24V Name Dry Pal Mode On Condition No 008 Stir Speed 1 80% Stir duration 20s Continue stirring No Condition No 009 Auxiliary instrument Control type Output24V Name Output_24V Mode Off Condition No 010 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence Yes Output sequence Start Wait for input sequence No Condition No 011 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence No Wait for input sequence Yes Maximum time 300 s Input sequence with results Yes Start sequence pH Total length 6 Number of results 1 Start position 1 3 Max. length 1 4 Condition No 012 Calculation R2 Name pH Unit Formula AuxInst[5] Coefficients No Decimal places 2 Result limits No Statistics No Condition No 013 Fill Automation InMotion Speed Low Filling mode Fixed duration Filling duration 30 s 014 Auxiliary instrument Control type USB-RS232 Name TV6 Send output sequence Yes Output sequence GOB\013 Wait for input sequence No Condition No 015 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence Yes Output sequence START2 Wait for input sequence No Condition No 016 Measure Cell Meas. temperature 20.00 °C Adjustment set Air&Water 20.00C Action when expired Warn Adj. temperature 20.00 °C Verify test set No Acquisition of measured values Wait time R 0 s Endpoint Automatic Meas. reliability Maximum Max. meas. duration 300 s Refracto eter ethod Setup 1 (incl. Degas): only required for degasing Application M677 METTLER TOLEDO METTLER TOLEDO - Page 12 of 14 - Automation in Food & Beverages Error detection Multiple measurement No 017 Calculation R1 Name nD Unit Formula nD Coefficients No Decimal places 5 Result limits No Statistics No Condition No 018 Calculation R3 Name Brix nD comp 20C Unit % w/w Formula T[Brix_ICUMSA_nD(nD)] Coefficients No Decimal places 2 Result limits No Statistics No Condition No 019 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence No Wait for input sequence Yes Maximum time ∞ s Input sequence with results Yes Start sequence AC Total length 8 Number of results 1 Start position 1 3 Max. length 1 6 Condition No 020 Calculation R4 Name ACID CONTENT WT% Unit wt% Formula AuxInst[8] Coefficients No Decimal places 3 Result limits No Statistics No Condition No 021 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence No Wait for input sequence Yes Maximum time ∞ s Input sequence CL Input sequence with results No Condition No 022 Auxiliary instrument Control type USB-RS232 Name TV6 Send output sequence Yes Output sequence GOA\013 Wait for input sequence No Condition No 023 Clean Automation InMotion Drain Yes Drain direction Waste Drain mode Fixed duration Drain duration 20 s Condition No 024 Online display Displayed results 4 Top left R1 Top right R2 Bottom left R3 Bottom right R4 025 Report Export to USB stick CSV + PDF file Export to network None Print / USB-RS232 data export Yes Print mode Summary Condition No 001 Title Type Measurement ID Setup 2 Title Acidity_Brix Author Administrator Date/Time 04/13/2015 Modified on 07/21/2015 Protect No SOP None 002 Configuration Cell mode R Automation InMotion Flex Lid handling No External instruments None 003 Sample Sample ID Cola Correction Factor 1.0 Comment No 004 Stir Speed 1 50% Stir duration 6s Continue stirring Yes Speed 2 35% Condition No 005 Auxiliary instrument Control type Output 24V Name DRY PAL Mode On Condition No 006 Stir Speed 1 80% Stir duration 20s Continue stirring No Condition No 007 Stir Speed 1 95% Stir duration 15s Continue stirring Yes Speed 2 25% Condition No 008 Auxiliary instrument Control type Output24V Name Output 24V Mode Off Condition No 009 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence Yes Output sequence Start Wait for input sequence No Condition No 010 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence No Wait for input sequence Yes Maximum time 300 s Input sequence with results Yes Start sequence pH Total length 6 Number of results 1 Start position 1 3 Max. length 1 4 Condition No 011 Calculation R2 Name pH Unit Formula AuxInst[4] Coefficients No Decimal places 2 Result limits No Statistics No Condition No Refractometer Method Setup 2 (incl. Degas): only required for degasing METTLER TOLEDO Application M677 METTLER TOLEDO - Page 13 of 14 - Automation in Food & Beverages 012 Fill Automation InMotion Speed Low Filling mode Fixed duration Filling duration 30 s 013 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence Yes Output sequence START2 Wait for input sequence No Condition No 014 Auxiliary instrument Control type USB-RS232 Name Tx titrator Send output sequence No Wait for input sequence Yes Maximum time ∞ s Input sequence with results Yes Start sequence AC Total length 8 Number of results 1 Start position 1 3 Max. length 1 6 Condition No 015 Measure Cell Meas. temperature 20.00 °C Adjustment set Air&Water 20.00C Action when expired Warn Adj. temperature 20.00 °C Verify test set No Acquisition of measured values Wait time R 0 s Endpoint Automatic Meas. reliability High Max. meas. duration 300 s Error detection Multiple measurement No 016 Calculation R1 Name nD Unit Formula nD Coefficients No Decimal places 5 Result limits No Statistics No Condition No 017 Calculation R3 Name Brix nD comp 20C Unit % w/w Formula T[Brix_ICUMSA_nD(nD)] Coefficients No Decimal places 2 Result limits No Statistics No Condition No 018 Calculation R4 Name ACID CONTENT WT% Unit wt% Formula AuxInst[8] Coefficients No Decimal places 3 Result limits No Statistics No Condition No 019 Clean Automation InMotion Drain No Drain direction No Condition No 020 PowerShower Beaker position Current position Initial drain No PowerShower rinse Yes Rinse cycles 1 Volume per cycle 35 mL Fill beaker No Condition No 021 Online display Displayed results 4 Top left R1 Top right R2 Bottom left R3 Bottom right R4 022 Report Export to USB stick CSV + PDF file Export to network None Print / USB-RS232 data export Yes Print mode Summary Condition No Application M677 METTLER TOLEDO METTLER TOLEDO - Page 14 of 14 - Automation in Food & Beverages Appendix: Complete list of accessories for this application: 1 T70 or T90 1 RM50 or RM40 1 51337150 Refractive index flow cell 1 51337029 Dry Pal 1 30094290 InMotion Autosampler Flex DERE 100mL 1 30094296 Aspiration tube (to use with DryPal for decarbonation) 1 51337197 Holding plate for pH/conductivity flow cell/DryPal 1 30094165 Pump SD660 Diaphragm w/Suction Tube 1 30094167 Tube Power Shower Rinse Flex 1 51109388 100 mL Beaker 120 pcs. 1 30094295 Adapter M8x1.5 2 51105856 USB-RS Adapter 1 51190362 RS 232C Cable 2 51109150 Compact Stirrer 1 30094247 Y-Cable pump/stirrer 2 30094237 SP280 Pump 1 51109220 Manual titration kit 1 22986 Clamping cone 1 51107501 Burette DV 1010 10mL 1 51108161 Aliquoting Beaker 100mL 1 51109504 pH-Sensor DGi115-SC 1 51109500 pH-Sensor DGi111-SC 2 51108034 Sensor-Cable 160cm 2 51108047 Tube Sample Loop 20cm 1 5110969 Novopren 1m (cut to length) 2 25961 Tubes 1 30026544 TV 6 2 51108049 Tube Sample Loop 2m (both ends with titration screwing) 1 11153130 LabX LiquiPhysics Express Starter Edition Pack optional 11153102 User management Centrally maintain and control users and roles within LabX. (included in Express & Server Packs) optional 11153103 Product data base Manage product data within LabX centrally – run methods based on product data optional 11153105 Auto Import/Export LabX allows you to import and export data into and from CSV and XML formats. METTLER TOLEDO Application M676 METTLER TOLEDO - Page 1 of 8 - Automation in Food & Beverages High Throughput Free and Total Sulfur Dioxide (SO2) Content in Wines An Italian testing lab with the help of Mettler-Toledo has developed a novel method to speed up the workflow of free and combined SO2 determination in wines using Titration Excellence and two InMotion Max Autosamplers. Preparation and Procedures CAUTION - Use safety goggles, a lab coat and wear gloves. If possible, work in a fume hood. - Ensure accurate cleaning of electrode is sufficient after each titration. Titer determination: - Weigh 0.1 g ascorbic acid in a 100 mL volumetric flask, add 30 mL deion. water to dissolve it. - Fill up to the mark with water and pipette 10 mL into the titration beaker for titer determination. - Add 50 mL deion. water, 5 mL 20 % H2SO4 and 10 mL 10 % KI solution. - Start immediately* the titer determination Sample preparation and titration free SO2 - Fill the titration beaker with 40 mL wine - 1 mL 20 % H2SO4 is added to the titration beaker. This step is performed using a pump - 5 mL 10 % KI solution are added. This step is performed using a pump - Start immediately* the determination of free SO2 Titration total SO2 - 6.4 mL 5 mol/L NaOH are added to liberate SO2 and let stand for 10 min to hydrolyse the combined SO2 - Further 3.7 mL 20 % H2SO4 are added to the titration beaker and determination is started. * Under strong acidic conditions, KI will convert to iodine with oxygen. Thus, the titration should start immediately after the KI addition. Remarks ‐ This application has been developed for an Italian testing lab according to their specific standards (stirring time, volume of H2SO4, KI solution and NaOH). ‐ It may be necessary to optimize the method for your sample and standards. ‐ Find further information about this success story in UserCom Anachem 20, case study of F. Lodroni. Sample 40 mL wine Compound Sulfur dioxide, SO2 M = 64.06 g/mol, z = 2 Chemicals 10 % potassium iodide, KI 20 % sulfuric acid, H2SO4 5 mol/L NaOH Titrant Iodine, I2 C(1/2 I2) = 0.025 mol/L Standard Ascorbic acid, C6H8O6, M = 176.13 g/mol, z = 2 Indication DM143-SC (voltametric indication) Platinum double-pin electrode Chemistry Titer determination: C6H8O6 + I2 → C6H6O6 + 2 I - + 2 H+ Free SO2 determination: SO2 + I2 + 2H2O → 4H+ + 2I- + SO4 2- Total SO2 determination: SO2 + I2 + 2H2O → 4H+ + 2I- + SO4 2- Calculation Content [mg/L]: Free SO2: R1=Q*C/m C=M*1000/z Combined SO2: R1=(Q[2]+QEX)*C/m C=M*1000/z Total SO2: R3=R1+R2 Waste disposal Neutralize the titrated solutions before final disposal Author, Version Francesco Lodroni, Area Manager Italy Cosimo De Caro & Julia Werder, MSG AnaChem, September 2015 Application M676 METTLER TOLEDO METTLER TOLEDO - Page 2 of 8 - Automation in Food & Beverages Instruments ‐ Titration Excellence T90 ‐ 2 x InMotion TM Max with PowerShower TM depending on the required throughput Accessories - LabX® Titration 2014 PC Software - 1 additional dosing unit (ME- 51109030) ‐ 2 x 10 mL DV1010 burette (ME-51107501) ‐ Titration Beaker 80 mL ‐ 8 x SD660 Diaphragm pump o 2 x Pump SD660 Diaphragm (ME- 30094165) for PowerShower ™ o 6 x Pump SD660 Diaphragm (ME- 30094165) for dosing of H2SO4, KI solution and NaOH METTLER TOLEDO Application M676 METTLER TOLEDO - Page 3 of 8 - Automation in Food & Beverages Results An Italian testing lab, the SO2 content of different wines from different wine producers are determined as single measurements. Therefore, no statistics can be provided. All results Method-ID ISVEA01 Sample ID Free SO2 Combined SO2 Total SO2 Consumption V [mL] Content [mg/L] Consumption V [mL] Content [mg/L] Content [mg/L] 1572202 0.110 3 0.751 21 24 1572425 0.108 3 0.526 15 18 1572601 0.761 21 1.753 48 69 1572834 0.565 16 2.971 82 98 1572835 1.178 33 3.718 103 136 1572865 0.614 17 1.773 49 66 1572866 0.424 12 1.618 45 57 1572869 0.074 2 0.868 24 26 1572870 0.128 4 1.033 29 33 1572871 0.164 5 0.892 25 30 1572872 0.127 4 0.720 20 24 1572876 0.080 2 0.402 11 13 1572877 0.177 5 0.509 14 19 1572878 0.076 2 0.442 12 14 1572879 0.188 5 0.763 21 26 1572880 0.156 4 0.470 13 17 1572881 0.150 4 0.642 18 22 1572882 0.088 2 0.488 13 15 1572883 0.186 5 1.139 31 36 1572884 0.087 2 0.578 16 18 1572885 0.055 2 0.362 10 12 1572890 0.805 22 2.747 76 98 1572896 0.899 25 2.856 79 104 1572913 0.464 13 0.824 23 36 1572917 0.538 15 1.012 28 43 1572920 0.362 10 1.558 43 53 1572921 0.956 26 3.245 90 116 1572923 0.805 22 3.493 96 118 1572926 0.167 5 0.760 21 26 1572929 0.172 5 0.755 21 26 1572931 0.143 4 0.871 24 28 1572932 0.236 7 0.620 17 24 1572945 0.336 9 1.130 31 40 1572953 0.802 22 2.130 59 81 1572954 0.819 23 1.586 44 67 1572956 0.049 1 0.390 11 12 Application M676 METTLER TOLEDO METTLER TOLEDO - Page 4 of 8 - Automation in Food & Beverages Titration curves Free SO2 Combined SO2 Table of measured values Sample ID 1572202 Free SO2: Volume Increment Signal Change Time Temperature mL mL mV mV s °C 0.0000 NaN 457.4 NaN 0 25.0 0.0000 0.0000 458.5 1.1 1 25.0 0.0010 0.0010 460.5 2.0 2 25.0 0.0040 0.0030 460.6 0.1 3 25.0 0.0090 0.0050 460.4 -0.2 4 25.0 0.0165 0.0075 454.7 -5.7 5 25.0 0.0275 0.0110 437.2 -17.5 6 25.0 0.0450 0.0175 372.3 -64.9 7 25.0 0.0665 0.0215 307.0 -65.3 8 25.0 0.0970 0.0305 152.0 -155.0 9 25.0 EP 0.109835 100.0 25.0 0.1165 0.0195 73.0 -79.0 10 25.0 0.1165 0.0000 92.0 19.0 11 25.0 0.1165 0.0000 96.6 4.6 12 25.0 Combined SO2: Volume Increment Signal Change Time Temperature mL mL mV mV s °C 0.0000 NaN 505.0 NaN 0 25.0 0.0000 0.0000 507.4 2.4 1 25.0 0.0010 0.0010 507.5 0.1 2 25.0 0.0040 0.0030 508.4 0.9 3 25.0 0.0090 0.0050 509.1 0.7 4 25.0 0.0160 0.0070 509.8 0.7 5 25.0 0.0280 0.0120 510.4 0.6 6 25.0 0.0430 0.0150 510.5 0.1 7 25.0 0.0665 … 0.4645 0.5485 0.6325 0.7165 0.0235 … 0.0755 0.0840 0.0840 0.0840 511.8 … 521.8 447.0 402.2 212.5 1.3 … 0.1 -74.8 -44.8 -189.7 8 … 15 16 17 18 25.0 … 25.0 25.0 25.0 25.0 EP 0.750990 100.0 0.7715 0.0550 33.1 -179.4 19 25.0 0.7715 0.0000 54.9 21.8 20 25.0 0.7715 0.0000 62.9 8.0 21 25.0 METTLER TOLEDO Application M676 METTLER TOLEDO - Page 5 of 8 - Automation in Food & Beverages Comments ‐ This method allows the determination of the free and total SO2 content in red wines by direct titration with iodine. The total SO2 content is defined as the sum of the free SO2 and the SO2 bound to other substances contained in the wine, i.e. aldehydes, ketones, etc. ‐ Reagents to be used:  Iodine is also reduced by other wine components. These competing reactions can partly be delayed by addition of 10 % KI solution.  The reaction only takes place in acidic medium, therefore H2SO4 must be added immediately before titration.  NaOH is added to hydrolyse the combined SO2 ‐ In order to measure both values from a single aliquot of sample, the following procedure is applied to each aliquot: 1. H2SO4 is dispensed 2. KI is dispensed 3. Titration of free SO2 4. NaOH is added to hydrolyse comined SO2 5. H2SO4 is dispensed 6. Titration of combined SO2 ‐ After the addition of NaOH in step 4, the reaction takes 10 min to hydrolyse the combined SO2 – this waiting time limits the throughput in standard methods. Together with Mettler Toledo an Italian testing lab developed an improved, automated method to avoid this waiting time:  Samples are grouped in several small series (usually 3 to 5 samples per series). For your interest: The grouping of a lot of samples can be done by an external program (e.g. excel) which automatically creates an import file for LabX.  On the first sample, steps 1 to 4 are performed. Then, instead of waiting for the hydrolysis to finish, the titrator immediately moves on to sample 2 and starts performing steps 1 to 4. This process, performing only steps 1 to 4, is repeated until the end of the series.  At the end of the series, the titrator returns to the first sample, checks the time from the NaOH addition and, when the required 10 min reaction time has been reached, steps 5 to 6 are performed. This process, performing steps 5 to 6, is repeated until the end of the series.  The next series can be programmed to proceed automatically, starting again with steps 1 to 4. Application M676 METTLER TOLEDO METTLER TOLEDO - Page 6 of 8 - Automation in Food & Beverages ‐ Some samples may contain ascorbic acid. These may lead to an increased detected amount of SO2, because of the reaction from ascorbic acid with iodine. ‐ In order to avoid loss of SO2, the samples must be taken from a freshly opened bottle. After opening the bottle, free SO2 evaporates or be oxidized while in contact with air resulting in too low results. ‐ To avoid binding of SO2 to organic compounds, a fast titration is required (2-3 minutes); particularly for wines with high SO2 content a lower value will be determined otherwise. Picture: Setup of the method for free and combined SO2 determination in wines at the Italian testing lab METTLER TOLEDO Application M676 METTLER TOLEDO - Page 7 of 8 - Sodium in Food & Beverages Methods 001 Title Type General titration Compatible with T90 ID ISVEA01 Title Free and total SO2 in wine Author METTLER TOLEDO Date/Time 14.08.2014 16:10:48 Modified at -- Modified by -- Protect No SOP None 002 Auxiliary value [1] Name SDS Formula H 0 Limits No Condition No 003 Sample (S1) Number of IDs 1 ID 1 Free SO2 Entry type Fixed volume Volume 40 mL Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C Sample data reader None 004 Titration stand [1] Type InMotion T/Tower A Titration stand InMotion T/1A Head position Sample Lid handling No 005 Stir Speed 30% Duration 5 s Condition No 006 Pump Auxiliary reagent H2SO4 Volume 1 mL Pump property 1-way Condition No 007 Pump Auxiliary reagent KI Volume 5 mL Pump property 1-way Condition No 008 Titration (EP) [1] Titrant Titrant ½ I2 Concentration 0.025 mol/L Sensor Type Polarized Sensor DM143-SC Unit mV Indication Voltametric Ipol 10 µA Temperature acquisition Temperature measurement No Stir Speed 30% Predispense Mode None Wait time 0 s Control Control User End point type Absolute Tendency Negative End point value 100 mV Control band 500 mV Dosing rate (max) 1 mL/min Dosing rate (min) 100 µL/min Termination At EP Yes Termination delay 2 s AT Vmax 10.0 mL Max. time infinite No Max. time 600 s Accompanying stating Accompanying stating No Condition Condition No 009 Calculation R1 Result Free SO2 (content) Result unit mg/L Formula R1 = Q*C/m Constant C=M*1000/z M M[sulfur dioxide] z z[sulfur dioxide] Decimal places 3 Result limits No Extra statistical func. No Send to buffer Yes Write to RFID None Condition No 010 Pump Auxiliary reagent NaOH Volume 6.4 mL Pump property 1-way Condition No 011 Rinse Auxiliary reagent Water Rinse cycles 1 Vol. per cycle 5 mL Position Current position Drain No Condition No 012 Auxiliary value [2] Name SDS Formula H H[SDS]+tUSE Limits No Condition No 013 End of sample 014 Sample (S2) Number of IDs 1 ID 1 Total SO2 Entry type Fixed volume Volume 40 mL Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C Sample data reader None 015 Titration stand [2] Type InMotion T/Tower A Titration stand InMotion T/1A Head position Sample Lid handling No 016 Stir Speed 30% Duration 600-(H[SDS]*60) s Condition No 017 Pump Auxiliary reagent H2SO4 Volume 3.7 mL Pump property 1-way Condition No 018 Stir Speed 30% Duration 10 s Condition No 019 Titration (EP) [1] Titrant Titrant ½ I2 Concentration 0.025 mol/L Sensor Type Polarized Sensor DM143-SC Unit mV Indication Voltametric Ipol 10 µA Temperature acquisition Temperature measurement No Stir Speed 30% Application M676 METTLER TOLEDO METTLER TOLEDO - Page 8 of 8 - Automation in Food & Beverages Predispense Mode None Wait time 0 s Control Control User End point type Absolute Tendency Negative End point value 100 mV Control band 30.0 mV Dosing rate (max) 125 mL/min Dosing rate (min) 100 µL/min Termination At EP Yes Termination delay 2 s AT Vmax 10.0 mL Max. time infinite No Max. time ∞ Accompanying stating Accompanying stating No Condition Condition No 020 Calculation R2 Result Fixed SO2 (content) Result unit mg/L Formula R1 = (Q[2]+ QEX)*C/m Constant C=M*1000/z M M[sulfur dioxide] z z[sulfur dioxide] Decimal places 3 Result limits No Extra statistical func. No Send to buffer No Write to RFID None Condition No 021 Calculation R3 Result Total SO2 (content) Result unit mg/L Formula R3=R2 + R1 Constant C=1 M M[sulfur dioxide] z z[sulfur dioxide] Decimal places 3 Result limits No Extra statistical func. No Send to buffer No Write to RFID None Condition No 022 Rinse Auxiliary reagent Water Rinse cycles 1 Vol. per cycle 5 mL Position Current position Drain No Condition No 023 Auxiliary value [3] Name SDS Formula H H[SDS]+tUSE Limits No Condition No 024 End of sample 025 Park Titration Stand InMotion T/1A Position Conditioning beaker Condition No 026 Record Report Report template Titration report Print No Condition No METTLER TOLEDO Application M674 METTLER TOLEDO - Page 1 of 6 - Automation in Food & Beverages Titrant concentration determination by automated volumetric KF titration Method for the volumetric Karl Fischer determination of the concentration determination of the titrant using the automated Karl Fischer Kit for InMotion™. Preparation and Procedures CAUTION ‐ Use safety goggles, a lab coat and wear gloves. If possible, work in a fume hood. Sampling ‐ Weigh 0.03 to 0.07 g disodium-tartrate dehydrate directly into a KF titration beaker ‐ Add a stirring bar, close the beaker with one aluminum foil and a rubber seal and put it on the InMotion™ rack Procedure ‐ Put two or three empty sealed beakers on the InMotion™ rack for the blank measurements ‐ The measurement of the blank is performed prior to the measurements of the samples ‐ 20 mL of methanol:formamide 1:1 (v/v) is added automatically to the beaker and the titration is performed Remarks ‐ The application method has been developed to be used with an InMotion™ Autosampler with automatic addition of solvent in the beaker ‐ The same stirring and measurement times were used for both the blank and the sample. This allows to include the drift into the Blank value, assuming that it is stable under controlled laboratory conditions ‐ The KF reagent and the solvent must be protected against intake of air humidity using drying tubes filled with molecular sieve. ‐ The mean value of the titer is automatically stored as part of the setup data by means of the function "Titer". ‐ The concentration of the KF titrant must be set to 5 mol/L since the concentration cannot be defined as mg H2O/mL. ‐ If the titer value is lower than 0.8, it is recommended to replace the titrant with fresh one. ‐ Pure methanol can also be used for the concentration determination. In this case, increase the stirring time from 240 to 400 s, in both blank and sample loops. Sample Disodium-tartrate dehydrate, 0.03-0.07 g Compound Water, H2O M=18.00 g/mol z=1 Chemicals HYDRANAL®Methanol Dry HYDRANAL®Formamide Dry Titrant One-component 5 mg H2O/mL (HYDRANAL®Composite 5 No.34805) Standard -- Indication DM 143-SC Double platinum pin electrode Chemistry CH3OH + SO2 + 3RN + I2 +H2O → (RNH)SO4CH3 + 2(RNH)I Calculation Blank: R1=VEND[1] [mL] Titer: R2=(m*15.66*10)/((VEND[2]- B[KF-kit])*c) m: mass of the sample (g) 15.66: % of water contained in disodium-tartrate dehydrate 10: Factor to have an unitless value VEND[2]: titrant consumption to EP (mL) B[KF-kit]: blank value c: concentration of the titrant (mg H2O/mL) Waste disposal Organic waste Author, Version Nicolas Monney, MSG AnaChem 2015, v1.0 Application M674 METTLER TOLEDO METTLER TOLEDO - Page 2 of 6 - Automation in Food & Beverages Instruments ‐ Titration Excellence T70/T90 ‐ XS205 Balance ‐ 1 x 10 mL DV1010 glass burette ‐ 1 x 20 mL DV1020 glass burette Accessories ‐ InMotion™ Flex ‐ Magnetic Stirrer (30094263) ‐ Aluminum foils (51108139) ‐ Rubber seal (51108298) ‐ KF titration beaker (51108125) Results All results Method-ID TiterKFKit Sample 0.05498 g (1/5) R3 1.00819 Sample 0.05426 g (2/5) R3 0.99673 Sample 0.05316 g (3/5) R3 1.03575 Sample 0.06179 g (4/5) R3 1.02617 Sample 0.05276 g (5/5) R3 1.06569 Statistics Method-ID TiterKFKit R3 Titer Samples 5 Mean 1.02651 s 0.02667 srel 2.597 % Titration curve Titer determination, sample 1/5 Titration curve Resutls METTLER TOLEDO Application M674 METTLER TOLEDO - Page 3 of 6 - Automation in Food & Beverages Table of measured values Comments  The rinsing unit of InMotion™ should be removed and the standard 9-hole insert head should be replaced by a special three hole plate (part number 30094163) that accommodates the electrode and the two dispensing tubes  The electrode and dispensing tubes should be mounted as shown on the following picture using the tube holder ring (part 51108083)  The dispensing tubes have to be placed above the two pins of the electrode, otherwise the piercing the aluminum foil might not be successful.  For this method to be compatible with T50, the blank measurement should be performed in a different method. In this case, remove the first sample loop and change "VEND[2]" for "VEND[1]" in the calculation R3. A blank measurement has to be performed before every sample series. Table of measured values Application M674 METTLER TOLEDO METTLER TOLEDO - Page 4 of 6 - Automation in Food & Beverages Methods 001 Title Type General titration Compatible with T70 / T90 ID TiterKFKit Title Titer determination with KF-Kit . . . 002 Sample Number of IDs 1 ID 1 Blank Entry type Fixed weight Weight 1.0 g Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C Entry Arbitrary 003 Titration stand Type InMotion T/Tower A Titration stand InMotion T/1A Head position sample Lid handling No 004 Stir Speed 40 % Duration 1 s Condition No 005 Dispense (normal) [1] Titrant MeOH_Formamide_1:1 Concentration 1 Volume 20 mL Dosing rate 60.0 mL/min Condition No 006 Stir Speed 40 % Duration 240 s Condition No 007 Titration (EP) [1] Titrant Titrant KF 1-component 5 Concentration 5 mol/L Sensor Type Polarized Sensor DM143-SC Unit mV Indication Voltametric Ipol 24 A Temperature acquisition Temperature measurement No Stir Speed 40% Predispense Mode None Wait time 0 s Control End point type Absolute Tendency Negative End point value 150 mV Control band 50 mV Dosing rate (max) 1 mL/min Dosing rate (min) 1 L/min Termination At EP Yes Termination delay 400 s At Vmax 10 mL Max. time infinite No Max. time 400 s Accompanying Stating Accompanying stating No Conditon Condition No 008 Calculation R1 Result Blank Result unit mL Formula R1=VEND[1] Constant C=1 M M[None] z z[None] Decimal places 3 Result limits No None Condition No Extra statistical func. No Send to buffer No Write to RFID None Condition No 009 End of sample 010 Calculation R2 Result Mean Blank Result unit mL Formula R2=Mean[R1] Constant C=1 M M[None] z z[None] Decimal places 3 Result limits No Send to buffer No Condition No 011 Blank Name KF-kit Value B=Mean[R1] Unit mL Limits No Condition No 012 Sample (Titer) Titrant KF 1-component 5 Concentration 5 mol/L Standard Disodium tartrate Entry type Weight Lower limit 0.03 g Upper limit 0.07 g Correction factor 1.0 Temperature 25.0 °C Entry Arbitrary Sample data reader None 013 Titration stand Type InMotion T/Tower A Titration stand InMotion T/1A Head position sample Lid handling No 014 Stir Speed 40 % Duration 1 s Condition No 015 Dispense (normal) [1] Titrant MeOH_Formamide_1:1 Concentration 1 Volume 20 mL Dosing rate 60.0 mL/min Condition No 016 Stir Speed 40 % Duration 240 s Condition No 017 Titration (EP) [1] Titrant Titrant KF 1-component 5 Concentration 5 mol/L Sensor Type Polarized Sensor DM143-SC Unit mV Indication Voltametric Ipol 24 A Temperature acquisition Temperature measurement No Stir Speed 40% Predispense Mode None Wait time 0 s Control End point type Absolute Tendency Negative End point value 150 mV Control band 50 mV METTLER TOLEDO Application M674 METTLER TOLEDO - Page 5 of 6 - Sodium in Food & Beverages Dosing rate (max) 1 mL/min Dosing rate (min) 1 L/min Termination At EP Yes Termination delay 400 s At Vmax 10 mL Max. time infinite No Max. time 400 s Accompanying Stating Accompanying stating No Conditon Condition No 018 Calculation R3 Result Titer Result unit -- Formula R3=(m"15.66*10)/((VEND[2]-B[KF-kit])*c Constant C=1 M M[Disodium tartrate] z z[Disodium tartrate] Decimal places 6 Result limits No Extra statistical func. No Send to buffer No Write to RFID None Condition No 019 End of sample 020 Titer Titrant KF 1-component 5 Concentration 5 mol/L TITER Mean[R3] Limits No Condition No 021 Calculation R4 Result Mean Titer Result unit Formula R4=Mean[R3] Constant C=1 M M[Disodium tartrate] z z[Disodium tartrate] Decimal places 5 Result limits No Send to buffer No Condition No Application M674 METTLER TOLEDO METTLER TOLEDO - Page 6 of 6 - Automation in Food & Beverages Picture: InMotion Autosampler with Karl Fischer Kit Picture: Karl Fischer Kit accessories: KF titration beaker, aluminum foil, a rubber seal and magnetic stirrers METTLER TOLEDO Application M675 METTLER TOLEDO - Page 1 of 6 - Automation in Food & Beverages Water content in food products determined by automated volumetric KF titration Method for the volumetric Karl Fischer determination of the water content of honey, jam and cocoa using the automated Karl Fischer Kit for InMotion™, together with the SmartSample™ workflow. Preparation and Procedures CAUTION ‐ Use safety goggles, a lab coat and wear gloves. If possible, work in a fume hood. Sampling ‐ Honey: weigh about 0.1 g directly into a KF titration beaker using a syringe without needle ‐ Jam: weigh about 0.04 g directly into a KF titration beaker using a syringe without needle ‐ Cocoa: weigh about 0.3 g directly into a KF titration beaker ‐ After weighing, store each weight on the RFID chip ‐ Add a stirring bar and close the beakers with one aluminum foil and a rubber seal and put them on the InMotion™ rack Procedure ‐ Put two or three empty sealed beakers on the InMotion™ rack for the blank measurements ‐ The measurement of the blank is performed prior to the measurements of the samples ‐ 20 mL of methanol:formamide 1:1 (v/v) is added automatically to the beaker and the titration is performed Remarks ‐ The application method has been developed to be used with an InMotion™ Autosampler with automatic addition of solvent in the beaker. ‐ The same stirring and measurement times were used for both the blank and the sample. This allows to include the drift into the Blank value, assuming that it is stable under controlled laboratory conditions. ‐ The KF reagent and the solvent must be protected against intake of air humidity using drying tubes filled with molecular sieve. ‐ The concentration of the KF titrant must be set to 5 mol/L since the concentration cannot be defined as mg H2O/mL. ‐ For sample dissolution, a stirring time of 240 s is needed for honey and jam and 360 s for cocoa. Sample Honey Strawberry jam Cocoa powder Compound Water, H2O M=18.00 g/mol z=1 Chemicals HYDRANAL®Methanol Dry HYDRANAL®Formamide Dry Titrant One-component 5 mg H2O/mL (HYDRANAL®Composite 5 No.34805) Standard Di-sodium tartrate dehydrate C4H4Na2O6.2H2O M=230.08 g/mol Indication DM143-SC Double platinum pin electrode Chemistry CH3OH + SO2 + 3RN + I2 +H2O → (RNH)SO4CH3 + 2(RNH)I Calculation Water content in %: (VEND[2]-B[KF-Kit])*c*TITER*C/m VEND[2]: titrant consumption to EP (mL) B[KF-Kit]: blank value c: concentration of the titrant (mg H2O/mL) TITER: titer of the titrant C: 0.1 (constant for % calculation) m: mass of the sample (g) Waste disposal Organic waste Author, Version Nicolas Monney, MSG AnaChem 2015, v1.0 Remarks Preparation and Prodedures Application M675 METTLER TOLEDO METTLER TOLEDO - Page 2 of 6 - Automation in Food & Beverages Instruments ‐ Titration Excellence T70/T90 ‐ XS205 Balance ‐ 1 x 10 mL DV1010 glass burette ‐ 1 x 20 mL DV1020 glass burette Accessories ‐ InMotion™ Flex ‐ Kit InMotion SmartSample (30094251) ‐ Magnetic Stirrer (30094263) ‐ Aluminum foils (51108139) ‐ Rubber seals (51108298) ‐ KF titration beakers (51108125) Results Honey samples: Results Method-ID Honey_KFKit Sample 0.08976 g (1/6) R3 17.637 % Sample 0.10478 g (2/6) R3 17.855 % Sample 0.09632 g (3/6) R3 17.640 % Sample 0.08407 g (4/6) R3 17.668 % Sample 0.09740 g (5/6) R3 17.690 % Sample 0.09895 g (6/6) R3 17.890 % Honey samples: Statistics Method-ID Honey_KFKit R3 Content Samples 6 Mean 17.730 % s 0.113 % srel 0.635 % Water content: Results of different samples Sample mass n Mean value Sdt Deviation srel [g] [% water] [% water] [%] Strawberry Jam 0.03 – 0.05 6 38.588 0.739 1.915 Cocoa powder 0.20 – 0.40 6 6.030 0.052 0.868 Resutls METTLER TOLEDO Application M675 METTLER TOLEDO - Page 3 of 6 - Sodium in Food & Beverages Titration curve Honey, sample 6/6 Table of measured values Titration curve Table of measured values Application M675 METTLER TOLEDO METTLER TOLEDO - Page 4 of 6 - Automation in Food & Beverages Comments  The SmartSample workflow is described on the scheme below:  For this method to be compatible with T50, the blank measurement should be performed in a different method. In this case, remove the first sample loop and change "VEND[2]" for "VEND[1]" in the calculation R3. A blank measurement has to be performed before every sample series. METTLER TOLEDO Application M675 METTLER TOLEDO - Page 5 of 6 - Automation in Food & Beverages Methods 001 Title Type General titration Compatible with T70 / T90 ID Honey_KFKit Title Honey with KF-Kit . . . 002 Sample Number of IDs 1 ID 1 Blank_Honey Entry type Fixed weight Weight 1.0 g Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C Entry Arbitrary Sample data reader None 003 Titration stand Type InMotion T/Tower A Titration stand InMotion T/1A Head position sample Lid handling No 004 Stir Speed 40 % Duration 1 s Condition No 005 Dispense (normal) [1] Titrant MeOH_Formamide_1:1 Concentration 1 Volume 20 mL Dosing rate 60.0 mL/min Condition No 006 Stir Speed 40 % Duration 240 s 360 s for cocoa Condition No 007 Titration (EP) [1] Titrant Titrant KF 1-component 5 Concentration 5 mol/L Sensor Type Polarized Sensor DM143-SC Unit mV Indication Voltametric Ipol 24 A Temperature acquisition Temperature measurement No Stir Speed 40% Predispense Mode None Wait time 0 s Control End point type Absolute Tendency Negative End point value 150 mV Control band 50 mV Dosing rate (max) 1 mL/min 1.5 mL/min for cocoa Dosing rate (min) 1 L/min 2 L/min for cocoa Termination At EP Yes Termination delay 400 s At Vmax 10 mL Max. time infinite No Max. time 400 s Accompanying Stating Accompanying stating No Conditon Condition No 008 Calculation R1 Result Blank Result unit mL Formula R1=VEND[1] Constant C=1 M M[None] z z[None] Decimal places 3 Result limits No Extra statistical func. No Send to buffer No Write to RFID None Condition No 009 End of sample 010 Calculation R2 Result Mean Blank Result unit mL Formula R2=Mean[R1] Constant C=1 M M[None] z z[None] Decimal places 3 Result limits No Send to buffer No Condition No Write to RFID None 011 Blank Name KF-kit Value B=Mean[R1] Unit mL Limits No Condition No 012 Sample Number of IDs 1 ID 1 Sample Entry type Weight Lower limit 0.0 g Upper limit 10 g Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C Entry Arbitrary Sample data reader RFID 013 Titration stand Type InMotion T/Tower A Titration stand InMotion T/1A Head position sample Lid handling No 014 Stir Speed 40 % Duration 1 s Condition No 015 Dispense (normal) [1] Titrant MeOH_Formamide_1:1 Concentration 1 Volume 20 mL Dosing rate 60.0 mL/min Condition No 016 Stir Speed 40 % Duration 240 s 360 s for cocoa Condition No 017 Titration (EP) [1] Titrant Titrant KF 1-component 5 Concentration 5 mol/L Sensor Type Polarized Sensor DM143-SC Unit mV Indication Voltametric Ipol 24 A Temperature acquisition Temperature measurement No Stir Speed 40% Predispense Mode None Wait time 0 s Application M675 METTLER TOLEDO METTLER TOLEDO - Page 6 of 6 - Automation in Food & Beverages Control End point type Absolute Tendency Negative End point value 150 mV Control band 50 mV Dosing rate (max) 1 mL/min 1.5 mL/min for cocoa Dosing rate (min) 1 L/min 2 L/min for cocoa Termination At EP Yes Termination delay 400 s At Vmax 10 mL Max. time infinite No Max. time 400 s Accompanying Stating Accompanying stating No Conditon Condition No 018 Calculation R3 Result Content Result unit % Formula R3=(VEND[2]-B[KF-Kit])*c*TITER*C/m Constant C=1/10 M M[None] z z[None] Decimal places 3 Result limits No Extra statistical func. No Send to buffer No Write to RFID None Condition No 019 End of sample 020 Calculation R4 Result Content Result unit % Formula R4=Mean[R3] Constant C=1 M M[None] z z[None] Decimal places 3 Result limits No Send to buffer No Condition No METTLER TOLEDO Application M657 METTLER TOLEDO - Page 1 of 6 - Automation in Food & Beverages Parallel Tasking: Sample Preparation and Titration with Two Towers This application allows to perform sample preparation and sample titration at the same time at Towers A and B of InMotion™ Autosampler: a sample is titrated for chloride content at Tower B while at the same time the next sample is prepared at Tower A. This results in a time saving of 45 - 50 %. Preparation and Procedures CAUTION ‐ Use safety goggles, a lab coat and wear gloves. If possible, work in a fume hood. ‐ Ensure that accurate cleaning of sensor is sufficient after each titration. Sample handling ‐ Homogenize the ketchup sample by e.g. mixing with a spatula sample before weighing. ‐ Put a beaker onto the balance ‐ Weigh accurately approx. 1 g of ketchup. Sample titration ‐ Place the sample beakers on the sample rack, together with conditioning and rinsing beakers filled with water, and an empty beaker at the end of the series. ‐ Start the method. ‐ 80 mL sulfuric acid is added automatically by means of a pump. ‐ Sample 1 is prepared at the 1 st tower (Tower A). In this case, the sample is stirred during 120 s. ‐ Subsequently, sample 1 is moved to tower B for titration, whereas sample 2 is moved to tower A for sample preparation. Remarks ‐ The application method has been developed for the mentioned sample. It may be necessary to optimize the method for your sample. - If necessary, the cleaning of the DM141i-SC electrode can be improved by using a diluted aqueous ammonia solution (ca. 1%) in the conditioning beaker. - To achieve accurate and precise results, samples are previously prepared at tower A before running the titration at tower B. In this case, stirring at 60% speed during was enough to completely dissolve the sample and release salt from ketchup. A heating step was not necessary. ‐ Literature: Titration Application M404 Sample Ketchup, 0.8-1.2 g Compound Chloride, Cl - M = 35.45 g/mol, z = 1 Chemicals 80 mL 0.02 M sulfuric acid (added by a pump) Titrant Silver nitrate, AgNO3 c(AgNO3) = 0.1 mol/L Standard Sodium chloride, NaCl, 0.03-0.05 g Indication DMi141-SC combined silver ring electrode (electrolyte: 1 M KNO3) Chemistry AgNO3 + NaCl = AgCl + NaNO3 Calculation Chloride content R = Q*C/m, in % C=M/(10*z) Waste disposal Filtration; precipitate (AgCl) has to be classified as special waste. Author, Version Matt Eby/Cosimo De Caro, MSG AnaChem, v1.0 Application M657 METTLER TOLEDO METTLER TOLEDO - Page 2 of 6 - Automation in Food & Beverages Instruments ‐ Titration Excellence T70/T90 ‐ 1 x 10 mL DV1010 glass burette ‐ XP205 Balance Accessories ‐ InMotion™ Autosampler Flex 100 mL (bundle: ME-30094120) with PowerShower™ ‐ Additional second tower ME-30094160 with PowerShower™ and SD660 pump ‐ CoverUp/Stirrer/Pump Option board ME-30094234 ‐ SYNC Y-cable ME-30094246 ‐ SP280 Peristaltic pump ME-30094237 ‐ PP Titration beakers ME-101974, spatula Results All results Method-ID M657 Loop 2/2 Sample 1/6 F&B sample (1/6) R1 (Content) 1.81 % Loop 2/2 Sample 2/6 F&B sample (2/6) R1 (Content) 1.79 % Loop 2/2 Sample 3/6 F&B sample (3/6) R1 (Content) 1.79 % Loop 2/2 Sample 4/6 F&B sample (4/6) R1 (Content) 1.80 % Loop 2/2 Sample 5/6 F&B sample (5/6) R1 (Content) 1.80 % Loop 2/2 Sample 6/6 F&B sample (6/6) R1 (Content) 1.79 % Statistics Method-ID M657 R1 Samples 6 Mean 1.80 % s 0.01 % srel 0.454 % Titration curve Sample 6/6 METTLER TOLEDO Application M657 METTLER TOLEDO - Page 3 of 6 - Automation in Food & Beverages Table of measured values Volume Signal Time mL mV s 0.0000 -20.9 0 0.2855 -20.6 3 0.4285 -20.0 6 0.5000 -19.8 9 0.6790 -19.5 18 1.0790 -17.5 22 1.4790 -15.2 24 1.8790 -12.0 28 2.2790 -8.4 31 2.6790 -3.8 34 3.0790 2.6 37 3.4790 11.0 40 3.8480 27.8 43 3.9285 34.0 46 3.9890 40.4 49 4.0445 47.7 52 4.0940 58.2 56 4.1175 64.9 59 4.1385 71.4 63 4.1615 81.2 66 4.1755 89.3 70 4.1855 96.4 73 4.1950 102.5 76 4.2085 111.9 79 4.2190 119.7 82 4.2290 126.3 85 4.2425 132.8 88 4.2650 142.4 92 4.2860 148.5 95 4.3275 157.7 98 4.3735 165.4 101 4.4365 173.1 104 Sample 6/6 Comments Resources: Type Name Stirrer Output Connection InMotion T/Tower A InMotion T/1A MB/STIRRER2 SYNC Y- cable Auxiliary instrument Tower A stirrer AB1/STIRRER Stirrer Tower A: connected with SYNC Y- cable InMotion T/Tower B InMotion T/1B MB/STIRRER1 Membrane SD660 InMotion T 1/Pump 1 Water – PowerShower Tower A Peristaltic SP280 InMotion T 1/Pump 2 H2SO4 – Tower A Membrane SD660_2 InMotion T 1/Pump 3 Water 2 – PowerShower Tower B Start: 1. Method M657 for sample preparation and titration contains two sample loops. 2. To start it, define a sample series of e.g. n = 6 samples as it follows: a. Loop 1: n = 1 Start InMotion T/1A: Position = 1 b. Loop 2: n = 6 All method functions “Titration Stand”: Start InMotion T/1A: Position = CP+1 3. In loop 2 the following parameters are entered: a. Sample IDs of all samples b. Sample mass of all samples Application M657 METTLER TOLEDO METTLER TOLEDO - Page 4 of 6 - Automation in Food & Beverages Principle: Salt content with parallel titration Comments Title – Parallel preparation Instruction Place empty cup after last sample – Time interval 5 s Sample n=1, fixed weight Titration Stand – Tower A Start position 1 Pump 0.1 M sulfuric acid – SP280 Stir Preparation 1 st sample Rinse Water – SD600 End of Sample Sample n=nTOT, IDs, Sample sizes Titration Stand – Tower A CP + 1 Pump 0.1 M sulfuric acid – SP280 Titration Stand – Tower B CP + 1 Auxiliary instrument - ON Tower A stirrer - preparation Stir Titration EQP – Salt Calculation R1 – Salt % Q*C/m, C=M/(10*z), M(NaCl) Stir – 0 % Stop stirrer Tower B Auxiliary instrument – OFF Titration Stand – Tower A CP + 1 Rinse Water – SD600 Conditioning Tower A Conditioning beaker Titration Stand – Tower B CP + 1 Conditioning Tower B Conditioning beaker End of Sample Park Tower A Conditioning beaker Park Tower B Current position METTLER TOLEDO Application M657 METTLER TOLEDO - Page 5 of 6 - Automation in Food & Beverages Methods 001 Title Type General titration Compatible with T70 / T90 ID M657 Title Parallel preparation+titration . . . 002 Instruction Instruction Place empty beaker after last sample and be sure that the temperature is correctly set at the DH100 Continue after Time interval Time interval 5 s Print No 003 Sample Number of IDs 1 ID 1 M657 Entry type Fixed Weight Weight 1.0 g Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C Sample data reader None 004 Titration stand (InMotion T/Tower A) Type InMotion T/Tower A Titration stand InMotion T/1A Titration head position Sample Lid handling No 005 Pump Auxiliary reagent H2SO4 Volume 80 mL Pump property 1-way 006 Auxiliary instrument (NOT USED since not needed) Control type Out TTL (Single pin) Name DH100 ON Mode Fixed time Time 2 s 007 Stir Speed 60_% Duration 120 s 008 Auxiliary instrument (NOT USED since not needed) Control type Out TTL (Single pin) Name DH100 OFF Mode Fixed time Time 2 s 009 Rinse Auxiliary reagent Water Rinse cycles 1 Vol. per cycle 10 mL Position Current position Drain No 010 End of sample 011 Sample Number of IDs 1 ID 1 F&B sample Entry type Weight Lower limit 0.7 g Upper limit 1 g Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C Sample data reader None 012 Titration stand (InMotion T/Tower A) Type InMotion T/Tower A Titration stand InMotion T/1A Titration head position Sample Lid handling No 013 Pump Auxiliary reagent H2SO4 Volume 80 mL Pump property 1-way 014 Auxiliary instrument (NOT USED since not needed) Control type Out TTL (Single pin) Name DH100 ON Mode Fixed time Time 2 s 015 Auxiliary instrument Control type Stirrer Name Tower A stirrer Mode On Speed 60 % 016 Titration stand (InMotion T/Tower B) Type InMotion T/Tower B Titration stand InMotion T/1B Titration head position Sample Lid handling No 017 Stir Speed 30_% Duration 10 s 018 Titration (EQP) [1] Titrant Titrant AgNO3 Concentration 0.1 mol/L Sensor Type mV Sensor DM141-SC Unit mV Temperature acquisition Temperature measurement No Stir Speed 30_% Predispense Mode Volume Volume 0.5 mL Wait time 5 s Control Control Normal Mode Precipitation Show paramaters No Evaluation and recognition Procedure Standard Threshold 200 mV/mL Tendency Positive Ranges 0 Add. EQP criteria No Termination At Vmax 10 mL At potential No At slope No After number of recognized EQPs Yes Number of EQPs 1 Combined termination criteria No Accompanying stating Accompanying stating No 019 Calculation R1 Result Content Result unit % Formula R1=Q*C/m Constant C=M/(10*z) M M[Chloride] z z[Chloride] Decimal places 2 Result limits No Record statistics Yes Extra statistical func. No Send to buffer No Condition None 020 Stir Speed 0_% Duration 120-20-(t*60) s 021 Auxiliary instrument (NOT USED since not needed) Control type Out TTL (Single pin) Name DH100 OFF Mode Fixed time Time 2 s Application M657 METTLER TOLEDO METTLER TOLEDO - Page 6 of 6 - Automation in Food & Beverages 022 Auxiliary instrument Control type Stirrer Name Tower A stirrer Mode Off 023 Titration stand (InMotion T/Tower A) Type InMotion T/Tower A Titration stand InMotion T/1A Titration head position Sample Lid handling No 024 Rinse Auxiliary reagent Water Rinse cycles 1 Vol. per cycle 10 mL Position Current position Drain No 025 Conditioning Interval 1 Position Conditioning beaker Time 10 s Speed 30 % Lid handling No 026 Titration stand (InMotion T/Tower B) Type InMotion T/Tower B Titration stand InMotion T/1B Titration head position Sample Lid handling No 027 Conditioning Interval 1 Position Conditioning beaker Time 10 s Speed 30 % Lid handling No 028 End of sample 029 Park Titration stand InMotion T/1A Position Conditioning beaker 030 Park Titration stand InMotion T/1B Position Current position 031 Record Summary Yes Results No Raw results No Resource data No Calibration curve No Method No Series data No METTLER TOLEDO Application M658 METTLER TOLEDO - Page 1 of 8 - Automation in Food & Beverages Parallel Titrations with Two Towers Methods M658A (client) and M658B (host) are synchronized to run parallel titrations at Towers A and B of InMotion™ Autosampler. Acid content (M658A) is determined at Tower A, while the chloride content (M658B) is determined at Tower B. First always start M658B, afterwards start M658A immediately. Preparation and Procedures CAUTION ‐ Use safety goggles, a lab coat and wear gloves. If possible, work in a fume hood. ‐ Ensure that accurate cleaning of sensor is sufficient after each titration. Sample handling ‐ Homogenize the ketchup sample by e.g. mixing with a spatula sample before weighing. ‐ Put a beaker onto the balance ‐ Weigh accurately approx. 1 g of ketchup. Sample titration ‐ Place the beakers on the rack, together with conditioning and rinsing beakers filled with water, and an empty beaker at the end of the series. ‐ Start the method: – Sample 1 is moved to Tower A. ‐ 50 mL water is added automatically by a pump at Tower A. Sample 1 is then titrated with NaOH. ‐ The result is stored as auxiliary value H[Acid] which is then used in method M658B for the calculation of the acidity content. ‐ Subsequently, sample 1 is moved to Tower B, where 10 mL sulfuric acid is added automatically by a pump, and it is titrated with AgNO3. ‐ At the same time, sample 2 is moved to Tower A for acidity titration in parallel. Remarks ‐ The method has been developed for this sample. It may be necessary to optimize the method for your sample. ‐ If necessary, the cleaning of the DM141i-SC electrode can be improved by using a diluted aqueous ammonia solution (ca. 1%) for conditioning in the special beaker 1. ‐ To achieve correct results, it is necessary that the sample is completely homogenized. In this case, stirring at 60% speed during at least 60 s was enough to completely dissolve the sample. - Literature: Titration Applications M400, M404 Sample Ketchup, 0.8-1.2 g Compound Acidic components – expressed as acetic acid, CH3COOH M = 60.05 g/mol, z = 1 Sodium chloride, NaCl M = 58.44 g/mol, z = 1 Chemicals 50 mL deionized water (added by a pump) 10 mL 0.02 M sulfuric acid (added by a pump) Titrant Sodium hydroxide, NaOH c(NaOH) = 0.1 mol/L Silver nitrate, AgNO3 c(AgNO3) = 0.1 mol/L Standard For NaOH: Potassium hydrogen phthalate, 0.08-0.12 g For AgNO3: Sodium chloride, 0.03-0.05 g Indication DGi115-SC (electrolyte: 3 M KCl) combined pH glass electrode DMi141-SC (electrolyte: 1 M KNO3) combined silver ring electrode Chemistry CH3COOH + NaOH = CH3COONa + H2O AgNO3 + NaCl = AgCl + NaNO3 Calculation Acid content, mg/g R1 = H[Acid]*C/m, C=M/z Salt content, mg/g R2 = Q*C/m, C=M/z Salt content, % R3 = Q*C/m, C=M/(10*z) Waste disposal Filtration; precipitate (AgCl) has to be classified as special waste. Author, Version Matt Eby/Cosimo De Caro, MSG AnaChem, v1.0 Application M658 METTLER TOLEDO METTLER TOLEDO - Page 2 of 8 - Automation in Food & Beverages Instruments ‐ Titration Excellence T90 ‐ 1 x additional DU Dosing unit ME- 51109030 ‐ 2 x 10 mL DV1010 glass burette ‐ XP205 Balance Accessories ‐ InMotion™ Autosampler Flex 100 mL (bundle: ME-30094120) with PowerShower™ ‐ Additional second tower ME-30094160 with PowerShower™ and SD660 pump ‐ CoverUp/Stirrer/Pump Option board ME-30094234 ‐ SYNC Y-cable ME-30094246 ‐ SP280 Peristaltic pump ME-30094237 ‐ PP Titration beakers ME-101974 ‐ Spatula Results All results Method-ID M658B Loop 2/2 Sample 1/6 F&B sample (1/6) R1 (Acetic acid content) 12.17 mg/g R2 (Salt content) 30.00 mg/g R3 (Salt content) 3.00 % Loop 2/2 Sample 2/6 F&B sample (2/6) R1 (Acetic acid content) 12.15 mg/g R2 (Salt content) 29.88 mg/g R3 (Salt content) 2.99 % Loop 2/2 Sample 3/6 F&B sample (3/6) R1 (Acetic acid content) 11.97 mg/g R2 (Salt content) 29.93 mg/g R3 (Salt content) 2.99 % Loop 2/2 Sample 4/6 F&B sample (4/6) R1 (Acetic acid content) 12.00 mg/g R2 (Salt content) 29.89 mg/g R3 (Salt content) 2.99 % Loop 2/2 Sample 5/6 F&B sample (5/6) R1 (Acetic acid content) 11.99 mg/g R2 (Salt content) 29.88 mg/g R3 (Salt content) 2.99 % Loop 2/2 Sample 6/6 F&B sample (6/6) R1 (Acetic acid content) 11.96 mg/g R2 (Salt content) 29.92 mg/g R3 (Salt content) 2.99 % Statistics Method-ID M658B R1 (Acetic acid content) Samples 6 Mean 12.04 % s 0.09 % srel 0.783 % R2 (Salt content) Samples 6 Mean 29.92 mg/g s 0.05 mg/g srel 0.153 % R3 (Salt content) Samples 6 Mean 2.99 % s 0.00 % srel 0.136 % METTLER TOLEDO Application M658 METTLER TOLEDO - Page 3 of 8 - Automation in Food & Beverages Titration curves M658A: acidity determination M658B: Chloride determination Table of measured values Volume Signal Time Volume Signal Time mL mV s mL mV s 0.0000 159.5 0 0.0000 -27.7 0 0.0050 159.3 2 0.0080 -28.3 3 0.0100 159.2 4 0.0160 -28.3 6 0.0225 158.7 6 0.0360 -28.5 9 0.0535 157.6 8 0.0860 -28.6 12 0.1310 154.8 10 0.2110 -28.3 15 0.3250 147.5 13 0.5235 -27.4 18 0.7940 130.1 16 0.9235 -26.1 21 1.2845 110.3 20 1.3235 -24.6 24 1.6965 88.0 24 1.7235 -22.8 27 1.9425 67.2 27 2.1235 -20.8 30 2.0800 50.0 31 2.5235 -18.7 34 2.1765 33.5 36 2.9235 -16.1 36 2.2540 16.2 40 3.3235 -13.5 40 2.3155 -0.6 45 3.7235 -9.9 43 2.3695 -18.2 50 4.1235 -6.1 46 2.4160 -35.3 55 4.5235 -1.0 49 2.4595 -52.3 59 4.9235 5.7 52 2.5025 -67.7 64 5.3235 16.8 55 2.5580 -84.0 68 5.5825 29.5 58 2.6325 -100.7 72 5.6885 39.4 62 2.7375 -118.2 76 5.7390 45.0 65 2.8825 -137.1 80 5.8055 55.4 69 3.0605 -155.5 84 5.8470 67.5 72 3.2785 -174.1 88 5.8635 73.1 75 3.5340 -191.6 91 5.8865 84.9 80 5.8980 92.3 84 5.9090 99.9 87 5.9215 109.0 90 5.9330 118.3 95 5.9430 125.4 99 5.9580 133.5 102 5.9795 142.5 106 6.0070 151.3 109 6.0445 158.7 112 6.1185 169.8 115 6.1970 177.8 118 6.3250 187.4 121 Acidity determination Chloride determination Application M658 METTLER TOLEDO METTLER TOLEDO - Page 4 of 8 - Automation in Food & Beverages CoCommments Resources: Type Name Stirrer Output Connection Auto stand Auto stand 1 AB1/STIRRER SYNC Y-cable InMotion T/Tower A InMotion T/1A MB/STIRRER2 Stirrer Tower A has to be connected using the SYNC Y-cable to two 24 V outputs InMotion T/Tower B InMotion T/1B MB/STIRRER1 Membrane pump SD660 InMotion T 1/Pump 1 Water – PowerShower Tower A Peristaltic pump SP280 InMotion T 1/Pump 2 H2SO4 – Tower B Membrane pump SD660_2 InMotion T 1/Pump 3 Water 2 – PowerShower Tower B Overview:  Workspaces A and B of Titration Excellence T90 are used: o Workspace A: Total Acid Content o Workspace B: Salt Content  Workspace A: o Total Acid Content - Method M658A (Client) o Acid/Base titration with 0.1 M NaOH and a DGi115-SC electrode o EQP Titration, 1 sample loop o Method function (MF) SYNC is used to synchronize with method M658B o Acidity result is stored as Aux. value H[Acid] for further use in method M658B  Workspace B: o Salt Content - Method M658B (Host) o Precipitation titration with AgNO3 and a DMi141-SC electrode o EQP Titration, 2 sample loops: - First loop: Synchronize acidity determination of 1st sample running on A by means of MF SYNC. - Second loop: Chloride titration and synchronization of acid content titration for subsequent samples. o The result stored under H[Acid] is called to calculate the acidity content o Acidity expressed as acetic acid content in mg/g (Calculation R1) o Salt content as mg NaCl/g and % NaCl (R2 and R3, respectively) Setup:  M658A (Client): - Enter total number of samples nTOT  M658B (Host) Loop 1 No. of samples n = 1 Start position InMotion T/1A: 1 Loop 2: No. of samples n = Total no. of samples Define IDs and sample mass of all samples 1st MF Titration Stand Start InMotion T/1A: CP+1 2nd MF Titration Stand Start InMotion T/1B: CP+1 3rd MF Titration Stand Start InMotion T/1A: CP+1 4th MF Titration Stand: Start InMotion T/1B: CP+1 Comments METTLER TOLEDO Application M658 METTLER TOLEDO - Page 5 of 8 - Automation in Food & Beverages Start:  First always start M658B (Host)  Subsequently, start immediately M658A (Client) Note: For easier use, define sample series and shortcuts for both methods. Methods for parallel titrations: Working Area A Working Area B Method A: Acidity Condition Comments Method B: Salt Condition Comments Title - Acid Title - Salt Sample - Fixed weight n = nTOT Instruction Place empty cup after last Titration Stand - Autostand sample - Time interval 5 s Sync - Wait 1 n ≤ nTOT Sample - Fixed Weight n = 1 Stir n ≤ nTOT Titration Stand - Tower A Start position 1 Titration EQP - Acid n ≤ nTOT Pump - Water PowerShower - SD660 1 Aux Value -> mmol Acid n ≤ nTOT H[Acid] = Q Sync - Send 1 Sync - Send 2 n ≤ nTOT Sync - Wait 2 End of Sample Rinse End of Sample Sample IDs, Sample sizes Titration Stand - Tower A CP+1 Pump - Water n < nTOT PowerShower Sync - Send 1 n < nTOT Calculation R1 -Acid mg/g H[Acid]*C/m, C=M/z Titration Stand - Tower B CP+1 Pump - H2SO4 H2SO4 , Peristaltic Stir Titration EQP - Salt Calculation R2 - Salt mg/g Q*C/m, C=M/z, M(NaCl) Calculation R3 - Salt % Q*C/m, C=M/(10*z) Stir = 0% Turn OFF stirrer Tower B Rinse Power Shower - SD600 2 Sync - Wait 2 n < nTOT Titration Stand - Tower A CP+1 Rinse n < nTOT Conditioning Tower A n < nTOT Conditioning beaker Titration Stand - Tower B CP+1 Conditioning Tower B Conditioning beaker End of Sample Park Tower A Conditioning beaker Park Tower B Current position Application M658 METTLER TOLEDO METTLER TOLEDO - Page 6 of 8 - Automation in Food & Beverages Methods Method M658A 001 Title Type General titration Compatible with T90 ID M658A Title Parallel titr. T/1A- Slave . . . 002 Sample Number of IDs 1 ID 1 Acid content Entry type Fixed Weight Weight 1.0 g Density 1.0 g/mL Correction factor 1.0 Temperature 25.0°C 003 Titration stand (Auto stand) Type Auto stand Titration stand Auto stand 1 004 Sync Action Wait Code 1 Condition Yes Formula n