Introduction

A spectrophotometer is designed to measure the amount of light absorbed by a substance. The instrument operates by passing a beam of light through the substance being investigated and measuring the intensity of light reaching a detector. http://www.chm.davidson.edu/vce/spectrophotometry/Spectrophotometry.html Spectrophotometry: a valuable technique which is especially useful when identifying compounds by determining their absorption of radiation in the visible light and UV regions. All coloured compounds absorb light at different wavelengths; this depends on two main principles: the concentration of the absorbing solute and the length of the path through absorbing solution. The beer-lambert law combines these two principles, it s=is usually expressed as follows: A=ECL (Absorbance=absorption coefficient. Concentration. Light path (cm)). The aim of the two experiments that will be carried out is to be able to understand how a spectrophotometer functions and to control it correctly to measure absorbance at different wavelengths; another aim is to calculate the absorption coefficient of a given substance using the beer lambert principle.

Method:
Experiment 1: Spectrophotometer is zeroed at wavelength 400 nm using distilled water as a blank, the absorbance of bromophenol blue and methyl orange is measured. Repeat the previous step at 20 nm intervals from 420 nm to 700 nm On the same graph plot absorbance against () If required take further readings to precisely locate ( max) Experiment 2: Dilute each compound to cover the concentration range 1 to 10 mg/l Set the spectrophotometer to wavelength of ( max), and zero against distilled water and measure the absorbance against each dilution of each compound. Plot absorbance against concentration for each compound on the same graph

1. Discussion
Explain why it was necessary to zero the spectrophotometer using distilled water. Comment on the difference in max of the two dyes. Do you think it relates to their chemical structures? Do you think your results have any errors and what would they be due to?