UMER MEHBOOB

06331413056
CONSTRUCTION OF
STANDARD CURVES
CONTENTS

 Definition
 Steps for construction of standard curve
 Serial dilution
 Properties of graph
 How to make a graph?
 Applications
DEFINITION

 It is a type of a graph used as quantitative research technique.

 The integrity of the quantitative assay depends upon the quality of the standard
reference.
 The sample with known properties is the standard reference and the graph is
known as standard curve.
STEPS FOR CONSTRUCTION OF
STANDARD CURVES

 To prepare the standard solutions of known concentration mainly by serial

dilution.
 Relate this known quantity to something we can measure.
 Graph the relationship between the two properties.
SERIAL DILUTION

 The process that reduces the concentration of solute in a solution in a geometric

way.
1. Selection of the liquid.
2. Preparation of glassware.
3. Preparation of the stock solution.
4. Preparation of various dilutions.
VARIABLE ON Y AXIS

 Mostly we plot standard curve by examining the absorbance of the sample

solution by a UV-visible spectrophotometer.
 A mobile phase is placed in one cuvette and the sample is placed in another
cuvette .
PROPERTIES OF GRAPH

 The standard curve between concentration and absorbance validates Beer

Lambert’s law.
 R2 is a number between 0.0 and 1.0.
 The nearer the value is to 1, the better the line of best fit in to the data and the
more linear the relationship is between the two variables.
 The validity of the graph depends upon the weighing, pipetting and handling of
the sample.
HOW TO MAKE A GRAPH
ON EXCEL SHEET ??
APPLICATIONS

 To determine unknown concentration of the compound.

 To analyze commercial products.
 To determine the concentration of radioactive compounds.
 To determine the amount of hydrocarbons present in water.
 Can estimate the number of microorganisms in a sample.
 To determine the amount of fertilizers in a soil.