LC/MS Analysis of Inositol Phosphates (VG-50 2D)

Phytic acid is a phosphoric acid ester of myo-inositol, in which six hydroxyl groups are replaced phosphate group, and it is also called myo-inositol hexaphosphate (IP6). In this application, phytic acid, IP5 (myo-inositol pentaphosphate) and IP4 (myo-inositol tetraphosphate) were simultaneously analyzed using HILICpak VG-50 2D, a polymer-based amino column with LC/MS detection. Alkaline conditions are suitable for high sensitive detection.

Sample: 5 µL
11,000-fold diluted Phytic acid
(ca.50 % in Water, ca.1.1 mol/L, Tokyo Chemical Industry Co., Ltd.)

  1. 1.Phosphoric acid
  2. 2.Phytic acid (IP6)
  3. 3.IP5
  4. 4.IP4
chromatogram of fructose and dextrose in cranberry fruit juice
Column
Shodex HILICpak VG-50 2D (2.0 mm I.D. x 150 mm)
Eluent
*0.1 M Ammonium carbonate buffer (pH10)/CH3CN=60/40
Flow rate
0.2 mL/min
Detector
ESI-MS (SIM Negative)
Column temp.
30 ℃

*Dissolve 7.9 g of ammonium bicarbonate in 980 mL of water, then add 20 mL of 25 % ammonia solution.

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