Measuring Absorbance Of Nitrophenol

In this science research laboratory, we measured the spectra of the protonated and ionized forms of p-Nitrophenol (PNP) by adding pH 5 and pH 10 buffer, respectively. The deprotonated sample ended up gift off a yellow color, and we measured two samples absorbances at contrastive wavelengths development a spectrophotometer. This lab also indispensable us to measure the pKa of PNP, by measuring the absorbance of many different samples of PNP + increasingly basic buffer samples, at the wavelength identified in the first experiment (~410nm). The wavelengths measured were from the visible range (350-800nm), and the spectrophotometer was blanked onward each measurement.
One of the main sciences covered in this lab was learning how to properly operate and utilize a spectrophotometer. This skill will be very important in the future, as many common assays (and future biochemistry labs) require quantification of the DNA/ribonucleic acid/protein in a sample at the end using spectroscopic methods. In this lab, I knowing how to use the spectrophotometer both by itself (manually) and by even outting up and using computing machine software to record data from the machine. I found taboo how to set the spectrophotometer software to automatically read all absorbances amidst a certain range of wavelengths, which was very convenient, compared to having to manually set the wavelength each time.



The computer program even measured the wavelengths in increments of 1nm, which was much smaller than the recommended increments of 10nm, which therefore gave my group much more detailed and precise data.
When using the machine, I also learned that if you are using a cuvette that has been used before, you should wipe it with a kimwipe before putting it into the machine. This step that makes sense, but it never occurred to me before, since I am used to using new, disposable cuvettes in my lab. This lab also taught me how to properly guide a cuvette, and that it is important to fill it with enough sample so that you can get a reading, but not too much, as...If you want to get a full essay, order it on our website: Orderessay



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