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1. matching the absorbance for determination2. Reagent blank E0 = coloration of reagent3. Slope of calibration curve for calculation4. Citation & unit (e.g. NO3-N mg/l)    Factors for citation & unit switch (e.g. NO3; mmol/l)5. Sample blank (e.g. coloration of the sample) is not included!   „Esample“ comes on top of E0 : individually for sample, for small volumes mostly negligible

No problem - our experts explain to you which two essential factors are important. Learn more about the suitability or the correct handling of the devices and sensors as well as the reagents.

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E = ελ· c · d ελ = molar absorptivity, in l/mol x cmd = Path length of the cell, in cmc = Concentration of the analyte, in mol/l

Lambert-Beer’s lawExperiments by BOUGUER (1698–1758) and LAMBERT (1728–1777) showed that the absorbance is dependent on the thickness of the absorbing layer of the cell used. The relationship between the absorbance and the concentration of the analyte in question was discovered by BEER (1825–1863). The combination of these two natural laws led to the derivation of Lambert-Beer’s law, which can be described in the form of the following equation:

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Filter photometers, e.g. photoLab® S12In lab with stable conditions, slow drift and often stabilizing reference beam requires less zeroing.

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Photometry is a measurement method to analyse (aqueous) solutions by means of a light source.Light (physical) is a spectra of electromagnetic waves, divided into different ranges: Visible light (white light) ranges from approx. 380 – 780 nm

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In our handbooks you will find our knowledge for your application in a compressed and clear form. The handbooks contain numerous application examples, tables, and diagrams that provide you with expert knowledge from years of research and development in a simple and clear manner.

Determination of substances by their specific colour reaction and light absorbance in dependance of their chemical properties at a specific wavelength.

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Methods/ programs in photometers contain all data and compute result automatically, including various cuvette sizes. Barcoded test kits additionally call up the respective method=program.

Absorbance = „Extinction of light“:Each substance has a specific spectra with absorbance peak(s).Run spectrum to define maximum or optimal peak = wavelength definition for concentration measurement

An overview of the high-precision photometers from WTW, a brand of Xylem Analytics Germany - universal, digital and portable spectrophotometers, high-precision robust filter photometers and the versatile portable photometers with multiparameter function for turbidity, pH and redox available - can be found here!

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1. Coloured solution contains dissolved dye2. Absorption of light leads to a coloration(complementary to λ)3. Color intensity is correlating with concentration4. The chemical reaction of analyte leads to building or disappearance (e.g. COD 4-40 mg/l) of dye in defined reaction time5. Reaction must be selective for the analyte – no cross reaction with other disturbing substances6. The developed dye must be stable for time of measurement => e.g. reading within 10 minutes after reaction time w/o color deterioration (see analytical instructions)

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Spectrophotometers, e.g. photoLab® 7000 SeriesZero/base line is required for many functions of spectral tasks, concentration mode is similar to filter photometers with reference beam.

Zeroing/Baseline:correcting the meter to „E0“ => especially after transport or in changing conditions (temperature…) meters E0 „drift“ some mAbs => readings become inaccurate (mostly too high)

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On 36 pages, you will find in the practical manual for photometry, in addition to many other tips and experience values of our experts for practice, for example, the 10 most important questions and answers about plausible measured values in photometry. Download now free of charge!

Photometric measurement procedures are among the most important measurement methods, along with pH measurement. They are used in research and teaching to investigate new substances or enzymatic and biochemical processes. In the environmental field, they are used to monitor environmental influences. In quality control they are used for food and beverages. The methodology ranges from absorption/transmission measurement and concentration determination to scanning and kinetics measurement. For decades, WTW has simplified the complex measurement process of photometry. For the user, this means convenient and fast measurement through automatic measurement settings and analytical quality control (AQC).

The correlation of absorbance/concentration is determined by setting up a characteristic calibration curve for each substance (parameter).The chemical reaction must be known:Dilution series with defined concentrations, measured at defined and cuvette size (pathlength)

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Konzentrationsmessung:Measurement at specific wavelength, obtained by either matching LED, optical filters from white light or mono-chromator

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Note: Scratches, pipette errors and other errors in the handling of the measuring instruments additionally influence the accuracy of the measurement results and lead to a higher inaccuracy!

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At the lower end detection limits and tolerances of the procedure have the biggest influence on accuracy of readings: Limitation of chemical procedure, confidence interval and accuracy are often at the lower limit.

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Click here to see all our Blog posts for Photometry Water Quality Product Support & How-to Sustainability Laboratory Agriculture Food and Beverage Industry

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Transmission measurement is also being used to measure turbidity at 180° angle (unit FAU, e.g. for quality control) and for turbidity correction in concentration measurement.

Performing a zero (see manuals!)LED meters, e.g. pHotoFlex® SeriesPortable meters require a zero due to changing conditions, transportation and optics.

The range depends on the equipment, the reaction has detection limits. MR-values are reaching approx. ± 2 - 2,5 Abs (test dependent!)

AQA Tools:1. Optical or liquid filters2. Color solutions, e.g. PhotoCheck®3. Selected unscratched zero cuvettes4. Control standards of the substance

1. Transmission T(%): Ratio of light intensity after cuvette (I) and before (Io)2. Absorbance: Abs = - log10 (T ) or „extinction of light“ passing the cuvette3. Concentration: quantitative analysis of a substance (mg/l, ppm,…) at a defined wavelength based on a calibration curve

100      10      1      0,1      Transmission (T%)               -            Logarithmic correlation0          1        2      3         Absorbance A = -log10 (T)    -            linear correlation0          4        8      12      Concentration (mg/l)              -            linear correlation

Influence of meter drift:  ZeroingPackage leaflet of many tests show sensitivity by correlating absorbance A (E=A) to mg/l.Influence can be seen directly:For COD test 14560, 4-40 mg/l COD, an absorbance of 10 mE means 0.4 mg/l COD. 10 mE drift without zeroing means 0.4 mg/l or 10% evitable miss-reading in the low end!

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