2. A. A crude dialysate yields an absorbance reading of 12 when undiluted. What
ID: 1085109 • Letter: 2
Question
2. A. A crude dialysate yields an absorbance reading of 12 when undiluted. What dilution would be necessary to get the absorbance of 0.5? 1:24 dilution needed B. Most cuvettes hold ~1 mL total volume and must be at least ¾ full to obtain a reading. If you wish to dilute the sample above, what's a reasonable volume to use for your diluted sample? C. Using the dilution factor you determine in part (A) and final volume chosen in part (B), what volume of the crude dialysate and what volume of buffer (to dilute) would you use?Explanation / Answer
2B)
net volume a cuvette can hold=3/4* 1ml=0.75ml
2C) dilution factor in part A)
Absorbance is directly proportional to concentration of absorbing species and so to the volume of the species
Absorbance1/Abs2=Volume1/volume2=12/0.5=24/1
dilution factor =1/24
To prepare 24 ml of diluted sample, 1ml undiluted sample is used
Thus, To prepare 0.75 ml of diluted sample, 1/24*0.75=0.0312 ml undiluted sample is used
0.0312 ml undiluted sample must be diluted to 0.75ml by adding buffer=0.75ml-0.0312ml=0.719 ml
Buffer=0.719ml
crude vol=0.0312ml
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