Why the assay of serum cystine by protein precipitation and chromatography should be abandoned.
The higher bias of serum cystine estimations by a HPLC method compared with those by ion exchange techniques is shown to be largely due to differences in the sample preparation procedures of the two techniques. The ion exchange methods utilised sulphosalicylic acid serum protein precipitation and post-column ninhydrin derivatisation of cystine, whilst the high pressure liquid chromatography technique employed automated dialysis for removal of proteins and pre-column ortho-phthalaldehyde derivatisation of cystine after its conversion to cysteine and then to S-carboxymethylcysteine. Examination of these procedures showed that whilst the high pressure liquid chromatographic method accurately estimates total serum cystine and cysteine, many factors affect the precision and accuracy of serum cystine estimations using the ion exchange techniques. In particular, serum protein precipitation techniques that are currently employed for the preparation of samples for cystine analysis by ion exchange chromatography should be abandoned.
Cooper
J D
JD
Biochemistry Department, Coventry and Warwickshire Hospital, Wiltshire, UK.
Turnell
D C
DC
Green
B
B
Wright
D J
DJ
Coombes
E J
EJ
D001798:Blood Proteins / Q000302:isolation & purification*; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D003553:Cystine / Q000097:blood*; D006801:Humans
D001798:Blood Proteins; D003553:Cystine