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Objective: To track the chymotrypsin-catalyzed hydrolysis of p-nitrophenyl acetate using the spectrometers.
Compound Molecular Formula Molecular Weight Boiling Point (°C) Melting Point (°C) Density (g/cm3)
p-nitrophenyl acetate C8H7NO4 181.14 -- 77 --
acetonitrile C2H3N 41.05 82 -45 0.786
Hydrochloric Acid HCl 36.46 110 -27.32 1.18
Sodium phosphate dibasic Na2HPO4 141.96 -- 250 0.5-1.2
Sodium phosphate monobasic NaH2PO4 119.98 -- -- --

Chemical Structures
p-nitrophenyl acetate



acetonitrile Hydrochloric Acid Sodium Phosphate dibasic
Sodium Phosphate monobasic





 
Observations:
For this lab, I was partnered up with 5 other people, should have been 3 but there were two extra in the class. First, we cleaned out 2 cuvettes to make sure they were free of chymotripsin. We left the bleach in the cuvettes for roughly 1 minute, rinsed them with DI water for another minute, and rinsed them with acetone so that they dry without too much trouble. The weight I was assigned was 15mg, the other weights were 5mg, 10mg, and 20mg.
Once one cuvette was dry, we proceeded to make a “zero” for the spectrometer. We added 2 portions of 940uL of the phosphate buffer and 100uL of p-nitrophenyl acetate. We used this sample to “zero” our spectrometer. Next, we each prepared our samples respectively and obtained the graph assigned to our concentration of chymotrypsin.
We used labtop 002, everything should be in the JABS folder.

Data:
Cleaning Cuvettes
Time bleach left in cuvette ~1 minute
Time rinsed with DI water ~1 minute
Preparation of Sample
Assigned weight 15mg
Amount HCl in microcentrifuge tube 1000uL
Amount phosphate buffer in cuvette -940uL =1880uL total
-940uL
Amount p-nitrophenyl acetate in cuvette 100uL
Amount...