Separation and identification of enzymatically prepared dephosphorylated products of myo‐inositolhexakisphosphate using LC‐MS |
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| Authors: | Purva Vats Bharat Bhushan Asit K. Chakarborti Uttam C. Banerjee |
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| Affiliation: | 1. Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, CT, USA;2. Department of Medicinal Chemistry, National Institute of Pharmaceutical Education and Research (NIPER), SAS Nagar, Punjab, India |
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| Abstract: | LC‐MS technique described here is a new way for the separation and direct determination of UV–Vis insensitive inositol phosphates (InsP2‐InsP6). This circumvents the need of radioisotopic labeling and post‐column derivatization techniques. The method involves separation of various enzymatically dephosphorylated derivatives of InsP6 on C18‐column using MeOH/H2O (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 InsP6 in a sequential manner leading to InsP2 (InsP2·2Na, tR 4.4–4.54 min, base peak m/z 382.9) as the end product. |
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| Keywords: | Inositol phosphates LC‐MS Phytase Phytic acid TLC |
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