High-Purity Separation of Phospholipids by Preparative High-Performance Liquid Chromatography

Duk Hui Kang; Kyung Ho Row

Journal of Industrial and Engineering Chemistry, Vol.7, No.3, 178-182, May, 2001

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High-Purity Separation of Phospholipids by Preparative High-Performance Liquid Chromatography

Duk Hui Kang, and Kyung Ho Row^†

Center of Advanced Bioseparation Technology and Dept. of Chem. Eng., Inha University, 253 Yonghyun-Dong, Nam-Ku, Inchon 402-751, Korea

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Soybean lecithin (phospholipids), mainly phosphatidylethanolamine(PE), phosphatidylinositol(PI), and phosphatidylcholine(PC), is a commercial by-product of vegetable oil production. The purpose of this work was to investigate the effects of packing size in a chromatographic column, and to obtain the optimum mobile phase composition for preparative work. Normal-phase High-Performance Liquid Chromatography (HPLC) was applied. Hexane, methanol, and isopropanol(IPA) were used as the ternary mobile phases. With the commercially available Nova-Pak column (4 In packing size, 3.9 mm i.d., 15 cm length), the gradient condition of mobile phase composition was that hexane linearly decreased from 85 to 0 vol.% and IPA and methanol, linearly increased 10 to 30 and 5 to 70 vol.% in the following two steps during the total run time of 25 min at the mobile phase flow rate of 1.5 mL/min. For the three other preparative packing sizes experimented were 15, 5/20, and 25/40 μm, the optimum mobile phases were experimentally obtained. The resolutions between Pi and PI, PI and PC with 15 μm packings was better over the other two preparative packings. A slight change in mobile phase composition was observed to increase the resolution at the same mobile phase flow rate.

Keywords:Phospholipid;Preparative HPLC;Gradient Condition;Packing Size

[References]

Olsson NU, Salem N, J. Chromatogr. B, 692, 245 (1997)
Munster C, Lu J, Schinzel S, Bechinger B, Salditt T, Eur. Biophys. J., 28, 683 (2000)
Hong Y, Pak Y, Phytochemistry, 51, 861 (1999)
Eckard PR, Taylor LT, Slack GC, J. Chromatogr. A, 826, 241 (1998)
Grizard G, Sion B, Bauchart D, Boucher D, J. Chromatogr. B, 740, 101 (2000)
Abidi SL, Mounts TL, J. Chromatogr. A, 773, 93 (1997)
Miwa H, Yamamoto M, Futata T, Kan K, Asano T, J. Chromatogr. B, 677, 217 (1996)
Christie WW, Lipid Analysis, 2nd ed., p. 107, Pergamom Press, New York (1992)
Miguel ID, Roueche A, Betbeder D, J. Chromatogr. A, 840, 31 (1999)
Bunger H, Pison U, J. Chromatogr. B, 672, 25 (1995)
Abidi SL, Mounts TL, J. Chromatogr. B, 741, 213 (1996)
Snyder LR, Kirkland JJ, Introduction to Modern Liquid Chromatography, 2nd ed., p. 34, Wiley, New York (1979)
Row KH, Lee JW, Korean J. Chem. Eng., 14(5), 412 (1997)
Row KH, Principles and Applications of Liquid Chromatography, p. 12, Inha Univ. (2000)

[Referenced By]

[Related Articles]

[1] Olsson NU, Salem N, J. Chromatogr. B, 692, 245 (1997)

[2] Munster C, Lu J, Schinzel S, Bechinger B, Salditt T, Eur. Biophys. J., 28, 683 (2000)

[3] Hong Y, Pak Y, Phytochemistry, 51, 861 (1999)

[4] Eckard PR, Taylor LT, Slack GC, J. Chromatogr. A, 826, 241 (1998)

[5] Grizard G, Sion B, Bauchart D, Boucher D, J. Chromatogr. B, 740, 101 (2000)

[6] Abidi SL, Mounts TL, J. Chromatogr. A, 773, 93 (1997)

[7] Miwa H, Yamamoto M, Futata T, Kan K, Asano T, J. Chromatogr. B, 677, 217 (1996)

[8] Christie WW, Lipid Analysis, 2nd ed., p. 107, Pergamom Press, New York (1992)

[9] Miguel ID, Roueche A, Betbeder D, J. Chromatogr. A, 840, 31 (1999)

[10] Bunger H, Pison U, J. Chromatogr. B, 672, 25 (1995)

[11] Abidi SL, Mounts TL, J. Chromatogr. B, 741, 213 (1996)

[12] Snyder LR, Kirkland JJ, Introduction to Modern Liquid Chromatography, 2nd ed., p. 34, Wiley, New York (1979)

[13] Row KH, Lee JW, Korean J. Chem. Eng., 14(5), 412 (1997)

[14] Row KH, Principles and Applications of Liquid Chromatography, p. 12, Inha Univ. (2000)