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Simple and Efficient Profiling of Phospholipids in Phospholipase D-modified Soy Lecithin by HPLC with Charged Aerosol Detection

  • Original Paper
  • Published:
Journal of the American Oil Chemists' Society

Abstract

Dietary phosphatidylinositol (PI) can be synthesized via phospholipase D (PLD)-catalyzed transphosphatidylation of phosphatidylcholine (PC), abundant in soy lecithin, with myo-inositol. However, a generated mixture of phospholipid (PL) classes poses a challenge for analysis. Our current work on Streptomyces PLD engineering requires a robust analytical method for profiling of PI and related PLs derived from the transphosphatidylation reactions. Therefore, we optimized an HPLC-based method with charged aerosol detector (CAD) for PL quantification. PLs were separated on a normal phase silica column by a gradient elution system using two solvents containing chloroform/methanol/1 M formic acid–triethylamine buffer in different ratios. Retention times of the PL standards and LC–MS under identical conditions were used to identity PL classes. PL standards gave linear response in 100- and 10-fold (lyso-PI) concentration range. The method provided a simple, sensitive, repeatable, and precise analysis of PI, PC, phosphatidylethanolamine, phosphatidic acid, and lyso forms of PC and PI. Compared to the similar existing method, introduction of CAD provided a three- to fivefold decrease at the lower end and a two- to fivefold increase at the upper end of the dynamic range. High precision, high sensitivity, and low limits of detection and quantification further underline the benefits of CAD in PL analysis.

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Acknowledgments

This work was supported in part by a Grant-in-Aid for scientific research C from Japan Society for the Promotion of Science to Y.I. [22560770]. The financial support to J.D. by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan is deeply appreciated.

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Authors and Affiliations

  1. Laboratory of Molecular Biotechnology, Department of Bioengineering Sciences, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8601, Japan

    Jasmina Damnjanović, Hideo Nakano & Yugo Iwasaki

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  1. Jasmina Damnjanović
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  2. Hideo Nakano
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  3. Yugo Iwasaki
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Correspondence to Yugo Iwasaki.

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Damnjanović, J., Nakano, H. & Iwasaki, Y. Simple and Efficient Profiling of Phospholipids in Phospholipase D-modified Soy Lecithin by HPLC with Charged Aerosol Detection. J Am Oil Chem Soc 90, 951–957 (2013). https://doi.org/10.1007/s11746-013-2236-x

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  • DOI: https://doi.org/10.1007/s11746-013-2236-x

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