Validated quantitative 31P NMR spectroscopy for positional isomeric impurity determination in L-α-glycerylphosphorylcholine (L-α-GPC)

Abstract

In this study a quantitative 31P nuclear magnetic resonance (31P NMR) spectroscopy method was described to determine positional isomeric impurity β-GPC in commercial products of L-α-GPC. The samples were dissolved in D2O and trimethyl phosphate (TMP) was selected as an internal calibrant. The measurements were performed on a Bruker 500 MHz spectrometer and the spectra were recorded under optimized process conditions. A good linear relationship was constructed for β-GPC in the range of 62.7–528.0 µg·mL−1, i.e. 0.03–0.25 % (w/w %, in relative to L-α-GPC) with a correlative coefficient of 0.9996. The limit of quantification (LOQ) and limit of detection (LOD) were 62.7 µg·mL−1 and 20.9 µg·mL−1 with signal to noise of 3 and 10, respectively. The spiked recoveries were in the range of 98.17–99.78 % with the relative standard deviation (RSD %) less than 1.0 %. Therefore, it could be supposed that the 31P NMR was a promising alternative method for sensitive determination of β-GPC for strict quality control of L-α-GPC.