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# Day 2 (pH Profile) Sequentially prepare the following mixtures in a glass cuvet:5aSb 2.006a6b7a7b 8a8b9a9bmL 0.10 M phosphate, pH 6.0 mL 0.10 M phosphate, pH 6.5 mL...

## Question

###### Day 2 (pH Profile) Sequentially prepare the following mixtures in a glass cuvet:5aSb 2.006a6b7a7b 8a8b9a9bmL 0.10 M phosphate, pH 6.0 mL 0.10 M phosphate, pH 6.5 mL 0.10 M phosphate, pH 7.0 mL 0.10 Mphosphate, pH 7.5 mL 0.10 M phosphate, pH 8.0 mL deionized water mL chymotrypsin stock (keep on ice) mL 0.0010 MHC2.002.002.002.002.002.002.002.002.000.500.500.500.500.500.500.500.500.500.500.400.400.400.400.400.400.400.400.400.40For each run, follow procedure for Day Steps 2-6 except use Biospec-180

Day 2 (pH Profile) Sequentially prepare the following mixtures in a glass cuvet: 5a Sb 2.00 6a 6b 7a 7b 8a 8b 9a 9b mL 0.10 M phosphate, pH 6.0 mL 0.10 M phosphate, pH 6.5 mL 0.10 M phosphate, pH 7.0 mL 0.10 Mphosphate, pH 7.5 mL 0.10 M phosphate, pH 8.0 mL deionized water mL chymotrypsin stock (keep on ice) mL 0.0010 MHC 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 0.50 0.50 0.50 0.50 0.50 0.50 0.50 0.50 0.50 0.50 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40 For each run, follow procedure for Day Steps 2-6 except use Biospec-1800 in kinetic mode to determine rates_ See the following page for parameter settings (CHY) The "b" runs serve as controls for the non-enzymatic hydrolysis of NPA, and are necessary because the rate of aqueous hydrolysis of NPA is strongly pH dependent:

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