calibretion curve

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CALIBRATION CURVE & SECONDARY DERIVATIVE CURVE:

By Dept. of Industrial pharmacy CALIBRATION CURVE & SECONDARY DERIVATIVE CURVE 1 gvrr416@gmail.com

contents:

contents Calibration curve Definition How to plot a curve Advantages & Disadvantages Applications Secondary derivative curve Definition Types 2 gvrr416@gmail.com

INTRODUCTION:

INTRODUCTION Calibration curve : It is a general method for determining the concentration of substance in an unknown sample by comparing the unknown to a set of standard samples of known concentrations. In this curve, concentration is taken on X-axis and absorbance is taken on Y-axis. This type of curves are generally seen in Spectroscopic studies. 3

HOW TO PLOT A CURVE:

HOW TO PLOT A CURVE 4

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We should prepare a series of standards across the range of concentrations near the expected unknown concentration. The concentrations of standards must lie within the working range of the technique(instrumentation) which we are using. Analysing each of these standards using the chosen technique will produce the series of measurements. 5

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For most of the analyzes a plot of instrument response VS analyte concentration will show a linear relationship. Thereby we can measure the response of the unknown and using the calibration can interpolate to find out the concentration of the analyte. 6

Advantages :

Advantages Most analytical techniques use calibration curve. This curve provides a reliable way to calculate the uncertainty of the unknown concentration. This provides the data on an empherical relationship . The mechanism for the instruments response to the analyte may be predicted. 7

Disadvantage:

Disadvantage Standards requires a supply of the analyte material preferably of high purity and in known concentration. 8 gvrr416@gmail.com

APPLICATIONS:

APPLICATIONS Analysis of concentrations. Verifying the proper functioning of an analytic instrument or a sensor device such as ion selective electrode. Determining the basic effects of a control treatment such as Dose survival curve in clonogenic assay 9

DERIVATIVE SPECTROSCOPY:

DERIVATIVE SPECTROSCOPY It involves the conversion of normal spectrum to its first , second or higher derivative spectrum. Types; Zeroth order First derivative spectrum Second derivative spectrum Higher derivative spectrum 10 gvrr416@gmail.com

SECONDARY DERIVATIVE CURVE:

SECONDARY DERIVATIVE CURVE This spectrum is a plot of the curvature of the zeroth spectrum against wavelength. or A plot of d 2 A/d λ 2 vs λ 11 gvrr416@gmail.com

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In the practical applications of spectrophotometry to the analysis of complex samples the spectral bands of the analyte are super imposed on a broad gradually curved background. Background of this type can be reduced by differentiation. This can be illustrated by following graphs 12

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13 Graph 1 Graph 2 gvrr416@gmail.com

summary:

summary 14 gvrr416@gmail.com

REFERENCES:

REFERENCES A. H. Beckett, Practical Pharmaceutical Chemistry, Fourth Edition- Part Two, CBS Publishers and distributors. www.google.com 15 gvrr416@gmail.com

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