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Separation and identification of enzymatically prepared dephosphorylated products of myo ‐inositolhexakisphosphate using LC‐MS
Author(s) -
Vats Purva,
Bhushan Bharat,
Chakarborti Asit K.,
Banerjee Uttam C.
Publication year - 2008
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200800372
Subject(s) - chemistry , chromatography , derivatization , mass spectrometry , resolution (logic) , hydrolysis , biochemistry , artificial intelligence , computer science
LC‐MS technique described here is a new way for the separation and direct determination of UV–Vis insensitive inositol phosphates (InsP 2 ‐InsP 6 ). This circumvents the need of radioisotopic labeling and post‐column derivatization techniques. The method involves separation of various enzymatically dephosphorylated derivatives of InsP 6 on C 18 ‐column using MeOH/H 2 O (30:70 v/v) and their identification using electron spray ionization MS in positive ion mode (+pESI‐MS). The LC‐MS studies revealed that the purified phytase from Aspergillus niger van Teighem hydrolyzes InsP 6 in a sequential manner leading to InsP 2 (InsP 2 ·2Na, t R 4.4–4.54 min, base peak m/z 382.9) as the end product.