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You are asked to separate two proteins via size exclusion chromatography. Your column is 5 cm in diameter and 25 cm in length. You add 50 g (dry weight) of Sephacryl gel (density = 1.25 g/cm³) to the column. The volume outside of the gel and pores is 77 cm³. The column operates at a flow rate of 1 mL/min to not compress the resin. The average retention times of proteins A and B are 244.5 and 273 min, respectively. Under the given conditions, you calculate that there are 1600 theoretical plates in the column. (This problem continues onto the next page). a) What is the volume inside of the pores? b) What are the elution limits of this system (i.e., what re the minimum and maximum volumes that particles will elute in)? c) What are the gel partition coefficients of proteins A and B for this system? d) What are the peak widths for both proteins? e) Assuming the peak widths are 22 mL and 28 mL for proteins A and B, respectively (these values are not necessarily correct), what is the resolution of the column?

You are asked to separate two proteins via size exclusion chromatography. Your column is 5 cm in diameter and 25 cm in length. You add 50 g (dry weight) of Sephacryl gel (density = 1.25 g/cm³) to the column. The volume outside of the gel and pores is 77 cm³. The column operates at a flow rate of 1 mL/min to not compress the resin. The average retention times of proteins A and B are 244.5 and 273 min, respectively. Under the given conditions, you calculate that there are 1600 theoretical plates in the column. (This problem continues onto the next page). a) What is the volume inside of the pores? b) What are the elution limits of this system (i.e., what re the minimum and maximum volumes that particles will elute in)? c) What are the gel partition coefficients of proteins A and B for this system? d) What are the peak widths for both proteins? e) Assuming the peak widths are 22 mL and 28 mL for proteins A and B, respectively (these values are not necessarily correct), what is the resolution of the column?

Introduction to Chemical Engineering Thermodynamics
Introduction to Chemical Engineering Thermodynamics
8th Edition
ISBN:9781259696527
Author:J.M. Smith Termodinamica en ingenieria quimica, Hendrick C Van Ness, Michael Abbott, Mark Swihart
Publisher:J.M. Smith Termodinamica en ingenieria quimica, Hendrick C Van Ness, Michael Abbott, Mark Swihart
Chapter1: Introduction
Section: Chapter Questions
Problem 1.1P
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You are asked to separate two proteins via size exclusion chromatography. Your column is 5 cm in diameter and 25 cm in length. You add 50 g (dry weight) of Sephacryl gel (density = 1.25 g/cm³) to the column. The volume outside of the gel and pores is 77 cm³. The column operates at a flow rate of 1 mL/min to not compress the resin. The average retention times of proteins A and B are 244.5 and 273 min, respectively. Under the given conditions, you calculate that there are 1600 theoretical plates in the column. (This problem continues onto the next page). a) What is the volume inside of the pores? b) What are the elution limits of this system (i.e., what re the minimum and maximum volumes that particles will elute in)? c) What are the gel partition coefficients of proteins A and B for this system? d) What are the peak widths for both proteins? e) Assuming the peak widths are 22 mL and 28 mL for proteins A and B, respectively (these values are not necessarily correct), what is the resolution of the column?
Transcribed Image Text:You are asked to separate two proteins via size exclusion chromatography. Your column is 5 cm in diameter and 25 cm in length. You add 50 g (dry weight) of Sephacryl gel (density = 1.25 g/cm³) to the column. The volume outside of the gel and pores is 77 cm³. The column operates at a flow rate of 1 mL/min to not compress the resin. The average retention times of proteins A and B are 244.5 and 273 min, respectively. Under the given conditions, you calculate that there are 1600 theoretical plates in the column. (This problem continues onto the next page). a) What is the volume inside of the pores? b) What are the elution limits of this system (i.e., what re the minimum and maximum volumes that particles will elute in)? c) What are the gel partition coefficients of proteins A and B for this system? d) What are the peak widths for both proteins? e) Assuming the peak widths are 22 mL and 28 mL for proteins A and B, respectively (these values are not necessarily correct), what is the resolution of the column?
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