I'm planing to analyze amino acids by HPLC. However, OPA-based method is complicated and does not reach the safety policy. Therefore, is there any other HPLC method for amino acids analysis? Previously, I used colorimetic using nihydrin but it was operated with amino acid analyzer not common HPLC system.
https://www.agilent.com/cs/library/slidepresentation/Public/Amino%20Acid%20Analysis_062410_Rita%20Steed.pdf
New approach for amino acid profiling in human plasma by selective fluorescence derivatization DOI: 10.1007/s00726-012-1243-9
http://www.fortis-technologies.com/resources/Fortis+Poster+HPLC+2012+Amino+Acids+v2.pdf
Article Standardised methods for amino acid analysis of food
I surely agree with you that OPA method is a complex method.
I would like to recommend the derivatization by PITC (phenylisothiocyanate; commecially available as Pico-Tag kit of Waters) and quantitative separation by our RP-HPLC method (please see file; PTH Roma RP-HPLC). This separation method does not use the notorious ion-pair reagents, and reliable.
Please, Cysteine should firstly be derivatized by using 4-vinylpyridine (Pyridil-ethylation (PE); standard PE-Cys is commercially available from MP Biomedicals though Sigma stopped selling) (please see file; PE derivatization).
You can synthesize PE-Cys from Cysteine according to the procedure as indicated in the file "PE derivatization". Synthesized PE-Cys can be purified and fractionated by using HPLC-photometric method. This fractionation may be achievable by using volatile eluent (please see file; Lipoyllysine Determin).
take a look at my latest poster on the analysis of Hypoglycin A in Ackee. It is one form of free amino acid. I used a mixed-mode column (Thermo Acclaim Q1) which will retain amino acid in an anion exchange mode.
Better yet, Imtakt has a new column for amino acid without derivatization. It was introduced at the ASMS meeting in San Diego three weeks ago.
Pico-Tag method seems to be the most popular method for amino acid analysis.
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