PRINCIPLES AND APPLICATION OF CHROMATOGRAPHY

PRINCIPLES AND APPLICATION OF CHROMATOGRAPHYAsheesh Pandey1

CHROMATOGRAPHY

Laboratory technique for the Separation of mixturesChroma -"color" and graphein - "to write.Colour bands - separation of individual compounds Measured or analysed.

Asheesh Pandey2

PURPOSE OF CHROMATOGRAPHYAnalytical Determine Chemical composition of a sample

Preparative Used to purify sufficient quantities of a substanceAsheesh Pandey3

TSWETT EXPERIMENT Asheesh Pandey4

Sample clean up is usually much less of a problem with HPLC than GLC and biological fluids can often be directly onto an HPLC column. Much sample pretreatment can also be avoided because aqueous solvents can be used in HPLC.because of all these advantages, HPLC has already made a significant impact in pharmaceutical, clinical, forensic and environmental analysis and it is now an ideal complementary technique to GLC.2. TYPES OF HPLC TECHNIQUES:A. Based on modes of chromatography1. Normal phase mode2.Reverse phase modeB. Based on principle of separation1. Adsorption chromatography2. Ion exchange chromatography3. Ion pair chromatography4.Size exclusion(or)Gel permeation chromatography5. Affinity chromatography6. Chiral phase chromatography

Asheesh Pandey5

C. Based on elution technique1. Isocratic separation2. Gradient separationD. Based on the scale of operation1. Analytical HPLC2. Preparative HPLCE. Based on the type of analysis1. Qualitative analysis2. Quantitative analysis3. PRINCIPLE: The principle of separation in normal phase mode and reverse phase mode is adsorption. When a mixture of components are introduced into a HPLC column, they travel according to their relative affinities towards the stationary phase. The component which has more affinity towards the adsorbent, travels slower. The component which has less affinity towards the stationary phase travels faster. Since no 2 components have the same affinity towards the stationary phase, the components are separated.Asheesh Pandey6

High-Performance Liquid Chromatography (HPLC)

Asheesh Pandey7

Chromatograph - equipment that enables a sophisticated separation EX. Gas chromatography or Liquid chromatographyEluent - Fluid entering column/ solvent that carries the analyte.Eluate - Mobile phase leaving the column.Stationary phase - Immobilized phase Immobilized on the support particles or on the inner wall of the column tubing.Examples : Silica layer - Thin Layer Chromatography

Asheesh Pandey8CHROMATOGRAPHY TERMS

Mobile phase Moves in a definite direction. Liquid (LC), Gas (GC).The mobile phase moves through the chromatography column (the stationary phase) where the sample interacts with the stationary phase and is separated.Retention time : Time takes for a particular analyte to pass through the system (from the column inlet to the detector) under set conditions. Sample (Anylate) :Substance analyzed in chromatography. Solvent : Any substance capable of solubilizing another substance.

Asheesh Pandey9

Chromatogram Visual output of the chromatograph. Separation - Different peaks or patterns on the chromatogram correspond to different components of the separated mixture.

Asheesh Pandey10

X- axis - Retention time

Y-axis - Signal

Signal is proportional to the concentration of the specific analyte separated.

Asheesh Pandey11

HOW TO DESCRIBE A CHROMATOGRAMAsheesh Pandey12

Asheesh Pandey13

Separation of PeaksAsheesh Pandey14

Retention Asheesh Pandey15k = (tr to)/ to

Where tr = the retention time of the compound, and to = the dead time

Higher values of k mean the analyte will stay in the column longer. The longer it stays, the more time there is for the peak will widen.

SelectivityAsheesh Pandey16a = kB/kA

the selectivity factor and is an indication of how well the compounds will separate. Higher means larger difference in retention time and more separation

EfficiencyAsheesh Pandey17The term that is generally used to describe column efficiency is number of theoretical plates or NN = L/H

Where: L =column lengthH = plate height (both in the same units)

N in Practical Terms...Asheesh Pandey18Units for tr and to.?Units for W1/2 ..?N can be measured from the peaks on a chromatogram..N = 5.54 trw1/2()2

ResolutionAsheesh Pandey19The purpose of chromatography is to separate or resolve compounds. The separation or distance between two peaks is known as their resolution and is a function of the 3 factors discussed previously: retention (the time it takes for the analytes to elute, related to k), selectivity (how different the analytes are from each other and related to ), and efficiency (how good the column is, related to N)

ResolutionAsheesh Pandey20Rs = (a-1/a) (k/k+1) NThe effect on Rs of:increasing a?increasing k?increasing N?EfficiencySelectivityRetention

ResolutionAsheesh Pandey21Rs = 2 (tR-B tR-A)/(wb-A + wb-B)

Where: A and B are the two peakstR = retention time andwb = the peak width at the base of each peak

Rs can also be calculated from actual measurements of peak retention times and measured peak widths

Retention Time: The time from the start of signal detection by the detector to the peak height of the elution concentration profile of each different sample.Curve Width: The width of the concentration profile curve of the different samples in the chromatogram in units of time.RESOLUTION (RS) :Rs = 2(tRB tRA)/(wB + wA)Where:tRB = Retention time of solute BtRA = Retention time of solute A wB = Gaussian curve width of solute B wA = Gaussian curve width of solute APlate Number (N):N = (tR)2/(w/4)2Plate Height (H):H = L/NWhere L is the length of the column.Asheesh Pandey22

ResolutionAsheesh Pandey23With a resolution value of 1.0, two peaks that overlap by about 4%. Values less than 1.0 indicate peaks that overlap, while at a resolution of 1.5, the peaks are considered fully separated.

Retention factor : R = Distance travelled by a Solute Distance travelled by a Solvent R = zero, - Solute remains in the stationary phase and thus it is immobile. R = 1 - Solute has no affinity for the stationary phase and travels with the solvent front. Asheesh Pandey24

Going back to N.Asheesh Pandey25N = L/HThe value of N is greatly dependent on the value of H.The value of H depends primarily on four factors:1) the velocity of the mobile phase,2) eddy diffusion or multipath diffusion, 3) the diffusion of the compound in the mobile phase4) the transfer of the compound between the stationary phase and the mobile phase.

H - Theoretical Plate HeightH = A + B/u + (Cs + Cm) uu = the average linear mobile phase velocity A is a term expressing multipath diffusionB/u is the term for longitudinal diffusionCs is the mass transfer term in the stationary phase Cm is the mass transfer term in the mobile phaseH = A + B/u + (Cs + Cm) uAsheesh Pandey26

A Multipath

12FlowDirection

Pathways of two molecules during elution. Distance traveled by molecule 1 is longer than that traveled by molecule 2, thus molecule 1 will take longer toelute.The amount of spreading is affected by the nature of the column material and how well the column is packed. This factor is generally proportional to the particle size of the packing material. This factor must be taken into account for packed columns, but for capillary columns, this term is not needed since there are no particles.Asheesh Pandey27

B Longitudinal Diffusion

Flow

Flow

Molecules diffuse from areas of highconcentration to areas of low concentration.

Over time.

At low velocities longitudinal diffusion has a negative effect on resolution, but this effect is negligible at higher velocities. This term is very important in gas chromatography as diffusion coefficients in gasses are orders of magnitude higher than in liquids. In liquid chromatography, this term is typically close to zero relative to the other terms.Asheesh Pandey28

Equilibrium between the mobile and stationary phases is never realizedMass Transfer Terms Cs & CmIt takes time for analytes to move from the mobile phase into the stationary phase. Because no equilibrium is reached, some of the analytes are swept ahead of the of the main band. It also takes time for molecules to move back out of the stationary phase, and some of the analyte molecules will be left behind by the rapidly moving mobile phase.Asheesh Pandey29

Mass Transfer Terms Cs & CmThe faster the mobile phase moves, the less time there is for equilibrium between the phases and the mass transfer effect on peak broadening is directly related to mobile phase velocity.Asheesh Pandey30

van Deemter PlotAsheesh Pandey31

Linear Velocity, uPlate Height, H

Multipath Term, A

Mass Transfer (both), CuLongitudinal diffusion, B/u

A + B/u + Cu

H = A + B/u + (Cs + Cm) u

PRICNIPLES OF CHROMATOGRAPGHY

Physical method of separation that distributes components to separate between two phases moves in a definite direction.

Substances are separated based on their differential distribution between two phases

Substances will move with the mobile phase at different rate depending upon their Partition or Distribution co-efficients.

Asheesh Pandey32

PRINCIPLESThe samples are subjected to flow by mobile liquid phase onto or through the stable stationary phase. Separation of fractions of mixture based on their relative affinity towards the two phases during their travel.The fraction with greater affinity to stationary phase travels slower and shorter while that with less affinity travels faster and longer.The separation is based on Differential partitioning between the mobile and stationary phases.

Asheesh Pandey33

Asheesh Pandey34

Asheesh Pandey35

FACTORES AFFECTING THE SEPARATIONIntermolecular interaction between the two phases

Extent of dispersion of solute molecules over the stationary phase

Asheesh Pandey36

CLASSIFICATION OF CHROMATOGRAPHYTechniques by Chromatographic bed shape Column chromatography Planar chromatography Paper chromatography Thin layer chromatographyTechniques by Physical state of mobile phase Gas chromatography Liquid chromatographyAffinity chromatography Supercritical fluid chromatography Asheesh Pandey37

TECHNIQUES BY CHROMATOGRAPHIC BED SHAPE

A.COLUMN CHROMATOGRAPHYPRINCIPLESSolid materials (Adsorbants) Ability to hold the molecules at their surfaceAttractive forces (Vanderwalls & Hydrogen )Functional groups (Hydroxyl/ Aromatic)Silica

Asheesh Pandey38

Stationary bed is within a tube. Solvent is driven through the column by applying Positive pressure.Separations - 20 minutes

Modern flash chromatography :Pre-packed plastic cartridges, Solvent is pumped through the cartridge. Quicker separations Less solvent usage.

Asheesh Pandey39

Column : Diameter - 5mm to 50mm Height - 5cm to 1 m with a tap Filter (a glass frit or glass wool plug)The individual components are retained by the stationary phase differently and separate from each other while they are running at different speeds through the column with the eluent. During the entire chromatography process the eluent is collected in a series of fractions. The composition of the eluent flow can be monitored and each fraction is analyzed for dissolved compounds, e.g., UV absorption, or fluorescence.

Asheesh Pandey40

STATIONARY PHASE

Asheesh Pandey41 Silica gel, Alumina. Cellulose

SOLVENTSHydroxyl groups - AlcoholCarboxyl group - AcetoneNon polar Compounds Hexane Heptane TouleneAsheesh Pandey42

Flow rate - Separation. Pump or compressed gas (e.g. Air, Nitrogen, Argon)A faster flow rate of the eluent: Minimizes the time required to run a column Minimizes diffusion Better separation.

Asheesh Pandey43

Asheesh Pandey44

Asheesh Pandey45

Asheesh Pandey46

B. PLANAR CHROMATOGRAPHYSeparation technique - Stationary phase is present as or on a plane.Paper Paper ChromatographyLayer of solid particles spread on a support such as a glass plate - Thin layer Chromatography.Different compounds in the sample mixture travel different distances according to how strongly they interact with the stationary phase as compared to the mobile phase. Retention factor (Rf)Asheesh Pandey47

47

Asheesh Pandey48

PAPER CHROMATOGRAPHYAsheesh Pandey49

Asheesh Pandey50

This paper is made of cellulose, a polar substance, and the compounds within the mixture travel farther if they are non-polar. More polar substances bond with the cellulose paper more quickly, and therefore do not travel as far.

Asheesh Pandey51PRINCIPLE

Asheesh Pandey52

Asheesh Pandey53

b) THIN LAYER CHROMATOGRAPHY

Widely employed laboratory technique Stationary phase - Adsorbent - Silica gel Alumina CelluloseWidely used in pharmaceutical & food stuff industry Advantages :Simple, Rapid and CheapFaster runs Better separationsChoice between different adsorbents. Better resolution Allow for quantification

Asheesh Pandey54

Used to identify the unknown compounds and to determine the purity of mixture.

TLC Plate - Aluminium or glass - coated by stationary phase.Coated material : 0.1-0.3mm in thickness Fluorescent indicator that will make it florescence during the UV light exposure.Asheesh Pandey55

MOBILE PHASE

Volatile Organic solventsAsheesh Pandey56STATIONARY PHASESilica gel, Alumina, or Cellulose on a flat, inert substrate.

Asheesh Pandey57

Asheesh Pandey58

SPRAYSAsheesh Pandey59

2.TECHNIQUES BY PHYSICAL STATE OF MOBILE PHASEA. GAS CHROMATOGRAPHYGas-Liquid chromatography, (GLC)Mobile phase Gas (Helium) Carrier Gas Pressure = 4 kg/cm2 Stationary phase - Column, which is typically "packed" or "capillary".The stationary phase is adhered to the inside of a small-diameter glass tube (a capillary column) or a solid matrix inside a larger metal tube (a packed column).Partition Coefficient of Volatile analyte between a solid stationary phase (Silicone) and a mobile gas (Helium).

Asheesh Pandey60

AdvantagesHigh sensitivity, High Resolution, High speedHigh Accurasy, Highly Quantitative APPARATUS Gas Chromatograph, GC analyzer, Normal syringes and one micro syringe, Beakers, Sample bottles and Electronic weight.

CHEMICALS Methanol, Isopropyl Alcohol and water

SAMPLE: Gases, Liquid, SolidsM.Wt: 2-800Volatile

Asheesh Pandey61

Asheesh Pandey62

APPLICATIONQuantitative & Qualitative analysis of low polarity compoundsAnalytical chemistry, Biochemistry, Petrochemical, Environmental monitoringMeasure picomoles of a substance in a 1 ml liquid sample, or parts-per-billion concentrations in gaseous samplesMeasuring toxic substances in soil, air or water. Asheesh Pandey63

APPLICATION OF GC- MSEnvironmental monitoring : Oraganic PollutantsCriminal forensics : Analyze the particles (Fibre) from a human body in order to help link a criminal to a crime.Law enforcement : Detection of illegal narcotics, Forensic toxicology : Find drugs and/or poisons in biological specimens of suspects, victims, or the deceased.Sports anti-doping analysis : Test athletes' urine samplesSecurity : Explosive detection (September 11 development) systems have become a part of all US airports.Food, beverage and perfume : from spoilage or Adultration - aromatic compounds, esters, fatty acids, alcohols, aldehydes, terpenesMedicine : Congenital metabolic diseases In Born error of metabolism

Asheesh Pandey64

B. LIQUID CHROMATOGRAPHYMobile phase - Liquid. Column or a plane. Very small packing particles and a relatively high pressure -High Performance Liquid Chromatography (HPLC).

Asheesh Pandey65

Asheesh Pandey66

Asheesh Pandey67

Asheesh Pandey68

Asheesh Pandey69

Asheesh Pandey70

LC- MSMass spectra is obtained rapidlySmall amount of material is required to form the spectra.Data collected is highly informative with respect to molecular structure.

Asheesh Pandey71

APPLICATION Pharmacokinetics : How quickly a drug will be cleared from the hepatic blood flow and organs of the body. Proteomics : Peptide mass fingerprintingDrug development: Peptide Mapping, Glycoprotein Mapping, Natural Products Dereplication, Bioaffinity Screening, In Vivo Drug Screening, Metabolic Stability Screening, Metabolite Identification, Impurity Identification, Degradant Identification, Quantitative Bioanalysis, and Quality Control.Fungal toxinsPesticides, HerbicidesAsheesh Pandey72

HIGH PERFORMANCE LIQUID CHROMATOGRAPHYHPLC V/S LC TECHNIQUEColumns : Small diameter (4.6 mm), stainless steel, glass or titanium. Column packing with very small (3, 5 and 10 m) particles Relatively high inlet pressures and controlled flow of the mobile phase.Detecting very small amountsHigh resolution Rapid analysisSpeed, efficiency, sensitivity and ease of operationHigh degree of versatilityEasily separate a wide variety of chemical mixtures400atmospheres. PUMP PRESSURE"Ultra High Performance Liquid Chromatography" systems 1000atmospheres.

Asheesh Pandey73

Asheesh Pandey74

ELUTION : Isocratic and Gradient.

ISOCRATIC :ISO ==> SAME- Solvent Composition Stays the Same for the Entire Run EX: 60:40 Alcohol:Water

GRADIENT : Solvent Composition Changes Throughout the Run

Asheesh Pandey75

Asheesh Pandey76

Asheesh Pandey77

Asheesh Pandey78

TYPES OF HPLCNature of the stationary phase Separation process

Adsorption chromatographyIon-exchange chromatographySize exclusion chromatographyAsheesh Pandey79

APPLICATIONProtein separationInsulin purificationPlasma fractionationEnzyme purificationAsheesh Pandey80

SIZE EXCLUSION CHROMATOGRAPHYGel filtration or gel permeation chromatography Separation - Molecular size of its components.Larger molecules are rapidly washed through the column, smaller molecules penetrate inside the porous of the packing particles and elute later.

Asheesh Pandey81

APPLICATIONS

Asheesh Pandey82

Asheesh Pandey83

AFFINITY CHROMATOGRAPHY

Based on specific & non-covalent binding of the proteins to other molecules Ligands ( His-tags, biotin or antigens)Physical properties of the analyte.

Biochemistry in the purification of proteins (Enzymes) bound to tags.

After purification, some of these tags are usually removed and the pure protein is obtained.

Asheesh Pandey84

SUPERCRITICAL FLUID CHROMATOGRAPHYUsed for the analysis and purification of low to moderatemolecular weight , thermally labile molecules. Principles are similar to those of(HPLC)Mobile phase - High pressure liquid orSuper critical Carbon Dioxide.Modifiers Methanol, Ehanol, isopropyl alcohol, acetonitrile and Chloroform.APPLICATIONUse in industry primarily for separation ofChiral (Asymmetric Carbon atoms) molecules.Serine Soman Glyceraldehyde Phosphours (Phosphine) Sulfar metal Cobalt Enkephalins

Asheesh Pandey85

DETECTORGas Chromatography or liquid Chromatography To visualize components of the mixture being eluted off the chromatography column.

Asheesh Pandey86

DETECTORSFlame ionization detectorAerosol-based detector Flame photometric detector (FPD). Atomic-emission detector (AED).Mass spectrometer ( MS) detectorNitrogen Phosphorus Detector, Evaporative Light Scattering Detector (ELD) : LC.

Asheesh Pandey87

DETECTORSUV detectorsThermal conductivity Detector, (TCD)Fluorescence detectorElectron Capture Detector, (ECD)Photoionization Detector, (PID)Refractive indexDetector (RI or RID)Radio flow DetectorChiral Detector

Asheesh Pandey88

Asheesh Pandey89

HPLC InstrumentationAsheesh Pandey90

90

In This Section, We Will Discuss:General components of a high performance liquid chromatograph.HPLC solvent delivery systems.How automatic injectors work.Common HPLC detectors.Asheesh Pandey91

91

HPLC Instrumentation OverviewAsheesh Pandey92

Principle PatternAn Example

DetectorThermostatted Column CompartmentAutosamplerBinary PumpVacuum DegasserSolvent CabinetSolvent ReservoirsController

Solvent FiltersSolvent Inlet FilerStainless Steel or glass with 10 micron porosity.Removes particulatesfrom solvent.Precolumn FilterUsed between the injector andguard column.2 to 0.5 micronRemoves particulates from sampleand autosampler wear debris.Must be well designed to preventdispersion.Asheesh Pandey93Guard column

Injector

Analytical ColumnPrecolumn Filter

Solvent Inlet Filter

Vacuum DegassingAsheesh Pandey94

Functions of the Solvent Delivery SystemThe solvent delivery system has three basic functions:

Provide accurate and constant flow.Provide accurate mobile phase compositions.Provide the force necessary to push the mobile phase through the tightly packed column.Asheesh Pandey95

Multichannel Gradient ValveDetermines mobile phase composition.

Largest solvent plug fills first.

Agilent 1100 and 1200 quaternary pump.96

Dual Piston Parallel PumpAsheesh Pandey97

SinglePistonDelivery

CombinedDelivery

Piston 'A' AdvancingPiston B Retracting

CheckValves

AB

PumpheadPistonRotarySwitchingValve

Dual Piston in Series PumpAsheesh Pandey98

First piston displaces solvent at twice the speed and stroke volume of the second piston.

Provides constant flow and the pressure necessary to get through column.

Ballvalves for Reciprocating Piston PumpsAsheesh Pandey99

Pump Seals and PistonsAsheesh Pandey100

PistonSupport RingsSeal KeepersSealsWear Retainers

2345

1

Frits and FiltersAsheesh Pandey101Frits, Filters, and Sieves are used to protect other parts of the LC from pump and seal material.

Purge valvePTFE Frit

Damping UnitsFilled with compressible liquid separated from the mobile phase by a membrane.Pressure ripples reduced to < 2% original value.102

Damping Unit

Pressure2% P/PPump Ripple

Gradient FormationAsheesh Pandey103

Low Pressure GradientHigh Pressure Gradient

SummaryThe pump is the most critical piece of equipment for a successfully operating HPLC.

Performance parameters for HPLC pumps:

Flow PrecisionFlow RangeDelay VolumePressure PulseComposition PrecisionAsheesh Pandey104

Sample InjectorsRequirements:

Reproducible introduction of the sample volume into the mobile phase flow.

Two major designs:Automatic Injectors or Manual InjectorsAsheesh Pandey105

Manual InjectorsAsheesh Pandey106

Front ViewInjectRear ViewLoad - Inject

Sample Loop

Manual InjectorsAsheesh Pandey107

Sample inSolvent inSolvent outSample LoadSample Inject

From PumpTo columnSample inSolvent inSolvent out

From PumpTo column

Automatic InjectorsAsheesh Pandey108

Step 1Step 2Step 3

Rotor SealsAsheesh Pandey109

Rotor Seal found withinvalve

Column OvenConstant temperature for solvent and column is required to perform reproducible results.110

Common HPLC DetectorsAsheesh Pandey111

UV-VISDiode ArrayMultiple WavelengthVariable Wavelength

Mass Spectrometers

Refractive Index

Fluorescence

Light Scattering

Electrochemical

Radioactivity

Conductivity

Necessity for More Than One Detector - SensitivityAsheesh Pandey112

Necessity for More Than One Detector - SelectivityAsheesh Pandey113

Necessity for More Than One Detector - Qualitative InformationAsheesh Pandey114Qualitative Information

Take peak spectrum (UV)

Chlortoluron ?

44685896132138158172215200Take peak spectrum (MS)104

Mass/ChargeAtrazine ?

Wavelength (nm)6080100120140160180200220

HPLC Detector CharacteristicsDetector performance characteristics: Sensitivity (LoD, LoQ) Selectivity Linearity Qualitative information Reliability Ease of use UniversalityAsheesh Pandey115

LODThe limit of detection for a detector can be characterized by its signal to noise ratio (S/N) for an analyte under a given set of conditions.Asheesh Pandey116

NoisePeak

Limit of Detection - Limit of QuantitationLimit of detection (LOD) is a result of the whole chromatography system, not only the detector performanceLimit of quantification (LOQ) is a defined limit for a method used for a specific purpose.117

Linear rangeSlope = sensitivityMQLMDLResponseAmountIntercepte.g.,RSD 20e.g., S/N > 3

UV-Vis DetectorsAsheesh Pandey118

bc

Detector Flow CellI0ILog I0 = A = abcI

Principles: The fraction of light transmitted through the detector cell is related to the solute concentration according to Beers Law.Characteristics: Specific, Concentration Sensitive, good stability, gradient capability.Special: UV-Vis Spectral capability (Diode Array Technology ).

UV-Vis Detectors - Design PrinciplesSingle wavelength detection of multi wavelength detection possible. Wavelength calibration is done automatically using a holmium filter.119

UV LampGratingFlow cellReference diodeSample diodeCut-off filterHolmium oxide filterSlitMirror 2Mirror 1

Variable WavelengthDetector

UV-Vis Detector with Spectral CapabilityDiode Array UV-Vis Detector allows online measurement of spectra.Wavelength range 190 - 950 nm. Wavelength Resolution: Up to 1 nm.Wavelength calibration with Holmium oxide filter.120

Diode ArrayGratingOpticalSlitDetectorFlow CellHomiumFilterAchromaticLensUVLampVisLamp

Fluorescence DetectionAsheesh Pandey121

Electrochemical DetectorsGold for carbohydrates.Platinum for chlorite, sulfate, hydrazine, etc.Carbon for phenols, amines.Silver for chloride, bromide, cyanide.122

Thin-layer design

Porous flow-through design

Wall-jet design

HPLC-MSD API- ElectrosprayAsheesh Pandey123

Refractive Index Detector DesignThe Refractive Index Detection is strongly influenced by:Pressure changesTemperature changes Flow pulseGradient elution is not possible!124

Conductivity DetectorsAsheesh Pandey125

ref.capacitorcellvariable resistancesfixed resistorCrBalancecontrol AEFDB

~Schematics

Applications

water

soap products

detergents

soft drinks

blood

plating baths

nuclear fuel reprocessing

streamsIonsAcidsBasesSaltsin}