Presentation on theme: "EC Line and the CL-10 Plus System Product Training"— Presentation transcript:
1EC Line and the CL-10 Plus System Product Training
2The EC- Line Product Range • CL-10 PLUSInstrument using the proprietary differential-pH technology.• EC- LineReagents. EC-Line is the range of reagents and diagnostics to be used with CL-10 instrument.
3The differential pH principle Let’s study the differential pH principle using the Urea Test in Milk.
4The components of the CL-10 Plus System Instrument:Two capillary glass electrodes to measure pHMixing chamberPeristaltic pumpsTubings that faciliate transfer of reagents and samples in the instrumentKits and supplies (EC-Line):(Urea Milk)Polif solution (Wash Solution)Regenerating SolutionStrong regenerating solution(Urea Kit) Reagents for the enzymatic reaction. Shelf life: 12 months
5Basic components of the instrument Left: Where solutions/samples get loaded into the instrumentRight: Where solutions/samples exit and enter the waste bottleWaste BottleResting: Polif SolutionTesting: R1 DiluentMixing chamberWhere milk samples are pipetted intoResting: Distilled WaterTesting: Urease Enzyme
6Pipetting into the reaction chamber Technique for pipetting into the reaction chamber.Load sample into pipetteInsert pipet into reaction chamberWait until you can feel the stirbar stirringDepress pipette all the way to release sample and keep it depressed as you lift the tip out of the chamber*This prevents fluid from being drawn out of the reaction chamber, affecting the reaction*Pipette tips are re-usable, wipe the outside of the pipette gently before inserting new samples
7The Urea ReactionUrease is an enzyme that catalyzes the hydrolysis of urea into carbon dioxide and ammonia.The ReactionUrea + 3 H2O 2NH4+ + CO2 + 2 OH-H2O + CO2 HCO3- + H+UreaseAs the reaction proceeds, the level of H+ will increase, the pH will decrease.As indicated in the reaction, the number of H+ ion is directly proportional to concentration of Urea.
9Enzyme-Urease being pumped to one of the electrodes (D2) Tempo (s)D1D2pHD2-D1dpH=aAc CoAD2Mixing ChamberEnzyme- Shown in Yellow being pumped into D2D1
10Starter (enzyme /substrate) The Reaction KineticsmpHBuffer + sampleStarter (enzyme /substrate)D1Enzymatic ReactionD2Time (s)
11What are D1, D2 and mpH?D1 = Difference in mpH measured by 2 electrodes filled with the same solution (buffer + sample)D2 = Difference in mpH measured by 2 electrodes when Electrode 1 has the same solution of D1 and Electrode 2 has D1 solution + enzymempH = Difference between D2 and D1mpH is driven by the enzymatic reaction.
12Why this technique? The technique is based on an enzyme reaction The measurement is not affected by interference from ammoniumCompared to traditional UV methods there is no color, opalescence and micro-particles interferenceHigh-tech micro-flow electrodes allow accurate, reproducible measurementsThese produce a repeatable, reliable, and accurate test method that has been adopted by many laboratories worldwideEliminate results subjectivity because there is no need to “interpret results”Test results are recorded and can be exported to Excel or printed out directly from the computer hooked up to the instrument
13Comparing with other techniques Traditional techniques:Titrators, refractometers, pH-meters Differential-pHSample treatment = No sample treatmentColour interference = No colour interferencePreservatives interference = No preservatives interferenceTime consuming = FasterOperator dependency = Operator independentLimited application panel = Wider application rangeAspecific = Highly specificTraditional techniques have lower specificity and lower accuracyRefractometers- Cannot differentiate between different sugarsAutomated pH meters and titratorsManual Titration
14Comparing with other techniques Spectrophotometers (UV) Differential pHSample treatment needed = No sample treatmentRather fast = Equally fast or slightly slowerShort linearity range = Wider linearityColour interference (auto-treatment) = No interferencePossible Matrix effects = No effect for tested matricesSome parameters not reliable = Reliabile for tested matricesAn autospectrometer
15Comparing with other techniques IR and FTIR Differential-pHNo sample treatment = no sample treatmentTime/test: test/hour = 15 sec/test to 5 minutesExpensive Instrument = Medium to lower cost instrumentsExpensive maintenance = Less expensive to maintainWineScan from FOSSMilkoScan from FOSSBentley For analysis of fat, protein and lactose in milk
16Comparing with other techniques HPLC Differential-pHRequires sample treatment = No sample treatmentVery accurate and precise = Equal and comparableTime/test 5 to 20 minutes = 15 sec/test to 5 minutesExpensive columns = Same equipment for all tests typesExpensive maintenanceDangerous waste = Disposable wasteUniversal use, versatile = Limited application/matrices panels
17Quality Indicators and Applications in Milk UreaLactoseL-Lactic AcidTitratable AcidityAlkaline Phosphatase
18Quality Indicators and Applications in Milk UREA (ISO Std 14637)In most countries urea is included in the milk payment scheme.Maximize feed levelsHigh urea concentrations high protein levelLow urea concentrations low protein levelUrea analysis in milk is important because it is related to the animal’s health.Levels of urea in milk stay in the range mg/100 cc.LACTOSELactose occurrs naturally in high concentrations in milk (≈130 mM).Also in the payment scheme, in some countries.In low-lactose milk products, this is also analysed for consumers with possible allergy/intolerance.
19Quality Indicators and Applications in Milk L-LACTIC ACIDIndicator of milk freshness. Fresh milk contains little to no Lactic acid. Milk for high quality production must contain maximum 30 ppm Lactic acid.Process control during fermentation in the yoghurt industries.In fermented milk and cheese products, the determination of D and L isomer forms of lactic acid is very important. Ratio affects taste and aroma.TITRATABLE ACIDITYIndicator for milk freshness.ALKALINE PHOSPHATASEAlkaline phosphatase enzyme is naturally found in milk. Its content in raw milk depends on breed, season and lactation phase of the animal.The enzyme is inactivated at the same temperature conditions where pathogens micro-organisms are destroyed.Its absence in milk is an indication of a successful pasteurization.
20Quality Indicators and Applications in Wine, Must and Cider pHSucroseGlucoseFructoseTotal acidityGlycerolCitric AcidAcetic AcidL-Lactic acidL-Malic Acid
21Quality Indicators and Applications in Wine GLUCOSE/FRUCTOSEControl of sugars in grapes and wine or must gives the alcoholic potential in final product.TOTAL ACIDITYAllows correction if de-acidification is needed.Allows monitoring of malic-lactic fermentation.Improves the taste of wine.pH also affects the color of wine.L-MALICMalic Acid is naturally present in must and develops into L-Lactic Acid in wine by an enzymatic reaction (Malolactic Fermentation).If not controlled, it could also turn into Acetic Acid, which is very dangerous!).L-LACTICMonitoring a balance between malic and lactic acid controls taste.Affects properties of the final product.
22Quality Indicators and Applications in Wine ACETIC ACID (the most abundant of volatile acids)An unwanted and dangerous compound.Corrections are possible at initial stages only!Above 1.08 g/l for white wines and 1.20 g/l for red wines, the product is not allowed to be sold as table wine.SUCROSE Sucrose does not naturally exist in wine, thus can be added for special fermentations (Champagne production) or as an Ethanol developer.
23CL-10 Instrument and EC Line Reagents Summary Comprehensive system and reagents for process and quality controlRequires no pre-sample treatmentReady-to-use reagentsFast and reliableNo sample centrifugationRapid AssayOfficial ISO-IDF method