Capillary Zone Electrophoresis에서 Deoxyribonucleotides의 분리

노경호

HWAHAK KONGHAK, Vol.33, No.2, 258-263, April, 1995

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Capillary Zone Electrophoresis에서 Deoxyribonucleotides의 분리

Separation of Deoxyribonucleotides by Capillary Zone Electrophoresis

노경호

초록

Capillary zone electrophoresis에서 organic modifier로서 isopropanol을 첨가하는 경우와 계면활성제로서 사용된 sodium dodecyl sulfate(SDS)의 농도에 따라서 4개의 deoxyribonucleotides(dAmp, dGmp, dTmp, dCmp)의 분리도에 미치는 영향을 고찰하였다. 실험기기는 60㎛ i.d fused silica tubing(68.5cm 길이), 고압 전원 공급기, UV 검지기(254nm)로 구성되어 있다. 최적의 이동상을 구하기 위해서 서로 조성이 다른 8개의 이동상에 따라서 분리도에 미치는 영향을 살펴보았다. 실험결과에 의하면 0.05M SDS, phosphate, Tris로 구성된 이동상이 가장 좋은 분리도를 보여 주었고 관의 효율(이론 단수)과 분리도는 분리전압에 의해서 영향을 받았다.

The effects of the addition of isopropanol as a organic modifier and the effects of concentration of sodium dodecyl sulfate(SDS) in capillary zone electrophoresis are discussed in terms of resolution using the four deoxyribonucleotides of dAmp, dGmp, DTmp, and dCmp. The system is consisted of a 60㎛ i.d. fused sillica tubing(68.5 cm column length), a high-voltage power supply, and an UV detector set at 254nm. The effects of the eight different mobile phases on the column efficiency and resolution were compared to find the optimum mobile phase. The experimental results showed that the mobil phase of phosphate/Tris with 0.05M SDS was most suitable for the separation of the components and the column efficiency(Number of theoretical plates)and resolution were affected by the separation voltage.

[References]

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[1] Jogerson JW, Phillips M, "New Directions in Electrophoretic Methods," ACS Symposium Series 335, ACS (1987)

[2] Lauer HH, McManigill D, Trend Anal. Chem., 5, 11 (1986)

[3] Jorgenson JW, Lukacs KD, Anal. Chem., 53, 1298 (1981)

[4] Balchunas AT, Sepaniak MJ, Anal. Chem., 59, 1466 (1987)

[5] Terabe S, Otsuka K, Ichikawa K, Tsuchiya A, Ando T, Anal. Chem., 56, 111 (1984)

[6] Armstrong DW, Sep. Purif. Methods, 14, 213 (1985)

[7] Row KH, Griest WH, Maskarinec MP, J. Chromatogr., 409, 193 (1987)

[8] Burton DE, Sepaniak MJ, Maskarinec MP, Chromatographia, 21, 583 (1986)

[9] Burton DE, Sepaniak MJ, Maskarinec MP, J. Chromatogr. Sci., 24, 347 (1986)

[10] Gehrke CW, McCune RA, Gama-Sosa MA, Ehrlich M, Kuo KC, J. Chromatogr., 301, 199 (1984)

[11] Perrin DD, Dempsey B, "Buffers for pH and Metal Ion Control," John Wiley & Sons, Inc., New York (1974)

[12] Cohen AS, Terabe S, Smith JA, Karger BL, Anal. Chem., 59, 1021 (1987)

[13] Maskarinec MP, Burton DE, Sepaniak MJ, "Selectivity of Micelles for Electrokinetic CHromatography," ORNL Internal Report (1987)

[14] Armstrong DW, Sep. Purif. Methods, 14, 213 (1985)