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hello everyone in this lecture today Iamp;#39;m going to talk to you about how to create this standard curve using observance and concentration and how to determine the concentration of unknown substance from the standard curve in Excel the first step is measuring the concentration of different standard samples okay so when we measure the concentration we measure at least two times and making technical replicates okay so the observance for each sample is measured two times okay so therefore we have absence one and observance two for example for the standard containing 0 microgram per ml concentration the absorbance one is 0.1 and absorbance 2 is 0.009 okay and similarly we will measure a two times constant observance for each of these consonant standards and we will get the different observance values okay absorbance one and absorbance two so after determining the after getting determining the observance value the next step is to calculate the average of these two values okay so then