Using the protocols you learned in Activity 1, calculate the amount of p-nitrophenol formed in all of your samples and record it in Table 16 in the third column.

Table 14. p-Nitrophenol standards

Standard	Amount of p-Nitrophenol (nmol*)	Absorbance 410 nm
S1	0	0
S2	12.5	0.206
S3	25	0.385
S4	50	0.831
S5	100	1.676

*1 nmol = 1 nanomole = 1 x 10–9 mol = 0.000000001 mol

Table 15. Determination of p-nitrophenol produced using a high and a low substrate concentration based on p-nitrophenol standards.

Cuvette	Standard That Is Most Similar	Amount of p-Nitrophenol Produced (nmol)
H1
H2
H3
L1
L2
L3

 

2. If you do not have a spectrophotometer, skip to Analysis of Results.

Quantitative Analysis of the Amount of Product Formed at Different Substrate Concentrations

1. Blank your spectrophotometer with the S1 standard at 410 nm. Measure the absorbance values for your six cuvettes and record the absorbance values in Table 16 in the second column.
2. Using the protocols you learned in Activity 1, calculate the amount of p-nitrophenol formed in all of your samples and record it in Table 16 in the third column.

Table 16. Determination of p-nitrophenol produced using a high and low substrate concentration based on a standard curve (similar to the one shown in Figure 8).

 

Cuvette	Absorbance at 410 nm	Amount of p-Nitrophenol Produced (nmol)
H1	0.584	37
H2	1.116	67
H3	2.500	150
L1	0.078	5
L2	0.289	20
L3	0.712	45

 

2.5 × 108 ÷ 1.8 = 150