Laser-based optical activity detection of amino acids and proteins

115 Pages·2016·1.64 MB·English

Iowa State University Capstones, Theses and
Retrospective Theses and Dissertations
Dissertations
1987
Laser-based optical activity detection of amino
acids and proteins
Barbara Hollenbeck Reitsma
Iowa State University
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Reitsma, Barbara Hollenbeck, "Laser-based optical activity detection of amino acids and proteins " (1987).Retrospective Theses and
Dissertations. 11721.
https://lib.dr.iastate.edu/rtd/11721
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Order Number 8721023
Laser-based optical activity detection of amino acids and
proteins
Reitsma, Barbara Hollenbeck, Ph.D.
Iowa State University, 1987
U M I
300N,ZeebRA
Ann Arbor, MI 48106
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Laser-based optical activity detection of
amino acids and proteins
by
Barbara Hollenbeck Reltsma
A Dissertation Submitted to the
Graduate Faculty in Partial Fulfillment of the
Requirements for the Degree of
DOCTOR OF PHILOSOPHY
Department: Chemistry
Major: Analytical Chemistry
Approved:
Signature was redacted for privacy.
In Charge o ajoi^ork
Signature was redacted for privacy.
Signature was redacted for privacy.
For the Grad e College
Iowa State University
Ames, Iowa
1987
11
TABLE OF CONTENTS
Page
GENERAL INTRODUCTION 1
Introduction to Amino Acids and Proteins 1
Chromatography of Amino Acids and Proteins 8
Laser-based Optical Activity Detector 13
SECTION I. HPLC DETERMINATION OF ENANTIOMERIC RATIOS OF
AMINO ACIDS WITHOUT CHIRAL SEPARATION 21
INTRODUCTION 22
EXPERIMENTAL 28
Chromatography 28
Optical Activity Detector 29
Procedure 30
Calculations 30
RESULTS AND DISCUSSION 31
REFERENCES 4(1
SECTION II. OPTICAL ACTIVITY AND ULTRAVIOLET ABSORBANCE DETECTION
OF DANSYL-L-AMINO ACIDS SEPARATED BY HPLC 48
INTRODUCTION 49
EXPERIMENTAL 51
Reagents 51
Chromatography 51
Detection 52
Calculations 53
RESULTS AND DISCUSSION 54
ill
REFERENCES 63
SECTION III. REVERSED-PHASE HPLC OF SOYBEAN TRYPSIN INHIBITOR WITH
OPTICAL ACTIVITY AND ULTRAVIOLET ABSORBANCE DETECTION 64
INTRODUCTION 65
EXPERIMENTAL ' 70
Reagents 70
Chromatography 70
Detection 71
Procedure 71
Calculations 72
RESULTS AND DISCUSSION 73
REFERENCES 9^
GENERAL SUMMARY 94
REFERENCES 99
ACKNOWLEDGEMENTS 104