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Difference Between Calibration Curve Absorbance and Concentration

June 25, 2018Posted byMadhu

Thekey differencebetween calibration curve absorbance and concentration is thatcalibration curve is a graph of absorbance and concentration, absorbance is the amount of light absorbed by a sample whereas concentration is the amount of a substance distributed in a unit volume.

Spectroscopyis an analytical technique which is useful in determining the concentration of an unknown compound present in a given sample. Therefore, it is aquantitative analysis. In this technique, we can determine the concentration of the compound using a curve. We should draw this curve between absorbance and the concentration. And we can draw the graph for several absorbance values obtained for different known concentration values. Then, we can use the absorbance value for the unknown sample to determine the concentration of that sample using the graph.

CONTENTS

1.Overview and Key Difference
2.What is Calibration Curve
3.What is Absorbance
4.What is Concentration
5.Relationship Between Calibration Curve Absorbance and Concentration
6.Side by Side Comparison – Calibration Curve Absorbance vs Concentration in Tabular Form
7.Summary

What is Calibration Curve?

A calibration curve is a standard graph which shows the change in the response of an analytical instrument towards different concentrations of the analyte. The analyte is the substance in which we need to find the concentration. In order to draw the calibration curve, we should use the known concentrations of the substance present in our sample to get a set of responses or signals. For spectroscopic techniques, the responses or signals are the absorbance values. Then, we can draw a graph between absorbance and concentration.

Difference Between Calibration Curve Absorbance and Concentration

Figure 01: The Structure of a Calibration Curve

We should draw the graph using the absorbance values obtained for each known concentration. We can use this graph to determine the unknown concentration of the same compound in a given sample by measuring the absorbance of that sample. The concertation value at the point of that absorbance value in the curve is the concentration of the compound in the sample.

What is Absorbance?

Absorbance is the response of a spectrophotometer towards the concentration of a sample. It measures the amount of light absorbs by the sample. This value depends on the amount and nature of the compound present in the sample. We can give this value using the following equation;

A = log(I/I0)

Where A is the absorbance, I is the intensity of the incident beam, and Iois the intensity of the transmitted beam through the sample. We can give this relationship in another way as follows:

A = –logT

Where T is thetransmittance. Therefore, the absorbance is related to the transmittance. for the same substance, if the concertation is high, the absorbance is also high and vice versa. That is because, if the concentration is high, the sample has a high amount of the compound that absorbs the light from the light beam. Moreover, when measuring the absorbance from a spectrophotometer, we should not use very high or very low concentrations. This is because, if we use very high concentrations, the intensity of the incident light beam may not sufficient for the total amount of compound present in the sample to absorb. If we use a low concentration, the sensitivity of the instrument may not sufficient enough to detect the low amount of compound in the sample.

What is Concentration?

Concentration is the amount of a substance that distributes throughout a unit volume of a sample. We can measure this in the unit of mol/L in which we give the amount of substance as a mole value and volume of the sample in liters. We call this the molar concentration. It is the most common unit for the measurement of the concentration.

Apart from that, we can determine the concertation as mass concentration, number concentration or a volume concentration.

t是什么he Relationship Between Calibration Curve Absorbance and Concentration?

We draw a calibration curve from two data sets: absorbance values and concentrations. We should take the absorbance as the y-axis and concentration as the x-axis because we can change the value of the concentration. Hence, it is the independent variable. But the absorbance changes according to the value of the concentration. Therefore, it is the dependent variable. Then, if we measure the absorbance of a sample, we can obtain the concentration of that sample using this calibration curve.

t是什么he Difference Between Calibration Curve Absorbance and Concentration?

A calibration curve is a standard graph which shows the change in the response of an analytical instrument towards different concentrations of the analyte. It indicates absorbance in y-axis and concentration in x-axis. Absorbance is the response of a spectrophotometer towards the concentration of a sample. It has no units. Concentration is the amount of a substance which distributes throughout a unit volume of a sample. Its unit is mol/L.

Difference Between Calibration Curve Absorbance and Concentration in Tabular Form

Summary – Calibration Curve Absorbance vs Concentration

Calibration curves, absorbance and concentration, are widely used terms in spectroscopy. The difference between calibration curve absorbance and concentration is that the calibration curve is a graph of absorbance and concentration and absorbance is the amount of light absorbed by a sample whereas concentration is the amount of a substance distributed in a unit volume.

Reference:

1. “Calibration Curve.” Wikipedia, Wikimedia Foundation, 15 June 2018.Available here
2. “Absorbance.” Wikipedia, Wikimedia Foundation, 15 June 2018.Available here

Image Courtesy:

1.’Calibration curve’By Kkmurray – Own work,(CC BY-SA 3.0)viaCommons Wikimedia

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Filed Under:Analytical Chemistry

About the Author:Madhu

Madhu is a graduate in Biological Sciences with BSc (Honours) Degree and currently persuing a Masters Degree in Industrial and Environmental Chemistry. With a mind rooted firmly to basic principals of chemistry and passion for ever evolving field of industrial chemistry, she is keenly interested to be a true companion for those who seek knowledge in the subject of chemistry.

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