정상 액체 크로마토그래피를 이용한 taxane으로부터 taxol의 분리

장경곤; 노경호; 정성택

Journal of the Korean Industrial and Engineering Chemistry, Vol.8, No.2, 286-291, April, 1997

Export Citation

정상 액체 크로마토그래피를 이용한 taxane으로부터 taxol의 분리

Separation of Taxol from Taxanes by NP-HPLC

장경곤, 노경호, 정성택

초록

항암제로 탁월한 효능이 있는 taxol을 정상 액체 크로마토그래피를 이용하여 주목나무의 추출물에서 분리하였다. Taxol을 불리하기 위해 시료 주입량은 5μl에서 100μl로 변화시키고, 이동상 유속은 1m1/min로 고정하여 일정 용매법으로 실험을 하였다. 실험에 사용된 이동상은 헥산, 에탄올, 메탄올, 1-프로판올, 이소프로판올이었다. Taxol과 유사한 성질를 갖는 표준시료 cephalomannine, taxol, 10-deacetyltaxol의 혼합물에서 taxol을 분리하였다. Cephalomannine, taxol, 10-deacetyltaxol 혼합물의 분리 실험 결과로 taxol의 분리 최적 조건은 2성분계에서 헥산/에탄올의 부피 조성비가 96/4이며, 3성분계에서는 헥산/1-프로판올/메탄올의 부피 조성비가 96/2/2이었다. 분리시간은 2성분계에서 80분인데 비해 3성분계에서는 50분으로 적은 용매를 사용하여 분리할 수 있었다. 위에서 얻는 실험조건으로 실제 주목나무의 추출물 1 ㎍에서 taxol를 분리할 수 있었다.

Taxol which is recognized as a powerful anticancer reagent was extracted from yew extract and separated by NP-HPLC (Normal-Phase High Performance Liquid Chromatography). The experiments were performed in the isocratic mode with 5μl to 100μl injection volume and 1ml/min mobile phase flow rate. The major mobile phase was hexane and small amounts of ethanol, methanol, 1-propanol and isopropanol were added to change the retention behavior. Prior to a real sample, the artificial mixture of taxol, cephalomannine and 10-deacetyltaxol was tested. They are hard to be separated because of similar chemical structures. The experimental results showed that the proper composition of mobile phase for separating the three components was found 96% hexane and 4% ethanol(vol.%) or 96% hexane, 2% 1-propanol and 2% methanol(vol.%). Compared to the run time of 80 minutes for the binary system, the mixture was separated within 50 minutes with the less amount of mobile phase for the ternary system. Finally, l㎍ of taxol was separated from yew tree extracts under the optimum operating conditions.

[References]

Wani MC, Taylor HL, Wall ME, Coggon P, McPhail AT, J. Am. Chem. Soc., 93, 2325 (1971)
Huizing MT, Rosing H, Koopman F, Keung ACF, Beijen JH, J. Chromatogr. B, 664, 373 (1995)
Wolff ME, "Burger's Medicinal Chemistry and Drug Discovery," 5th ed., John Wiley & Sons, NY, U.S.A., 996 (1995)
Harvey SD, Campbell JA, Kelsey RG, Vance NC, J. Chromatogr. B, 664, 300 (1991)
Ketchum REB, Gibson DM, J. Liq. Chromatogr., 16, 2519 (1993)
Witherup KM, Look SA, Stasko HW, Mccloud TG, Muschik HJ, J. Liq. Chromatogr., 12, 2117 (1989)
Appendino G, Tagliapietta S, Ozen HC, Gariboldi P, Gabetta B, Bombardelli E, J. Nat. Prod., 56(4), 514 (1993)
Ketchum REB, Gibson DM, J. Liq. Chromatogr., 18, 1093 (1995)
Wu DR, Lohse K, Greenblatt HC, J. Chromatogr. A, 702, 234 (1995)
Castor TP, Tyler TA, J. Liq. Chromatogr., 16, 724 (1993)
Cardellina JH, J. Liq. Chromatogr., 14, 661 (1991)
Hamilton RJ, Sewell PA, "Introduction to High Performance Liquid Chromatography," 2nd Ed., Chapman and Hall, NY, U.S.A., 29 (1982)
Kopycki WJ, Elsohly HN, Mc-Chesney JD, J. Liq. Chromatogr., 17, 2569 (1994)

[Referenced By]

[1] Wani MC, Taylor HL, Wall ME, Coggon P, McPhail AT, J. Am. Chem. Soc., 93, 2325 (1971)

[2] Huizing MT, Rosing H, Koopman F, Keung ACF, Beijen JH, J. Chromatogr. B, 664, 373 (1995)

[3] Wolff ME, "Burger's Medicinal Chemistry and Drug Discovery," 5th ed., John Wiley & Sons, NY, U.S.A., 996 (1995)

[4] Harvey SD, Campbell JA, Kelsey RG, Vance NC, J. Chromatogr. B, 664, 300 (1991)

[5] Ketchum REB, Gibson DM, J. Liq. Chromatogr., 16, 2519 (1993)

[6] Witherup KM, Look SA, Stasko HW, Mccloud TG, Muschik HJ, J. Liq. Chromatogr., 12, 2117 (1989)

[7] Appendino G, Tagliapietta S, Ozen HC, Gariboldi P, Gabetta B, Bombardelli E, J. Nat. Prod., 56(4), 514 (1993)

[8] Ketchum REB, Gibson DM, J. Liq. Chromatogr., 18, 1093 (1995)

[9] Wu DR, Lohse K, Greenblatt HC, J. Chromatogr. A, 702, 234 (1995)

[10] Castor TP, Tyler TA, J. Liq. Chromatogr., 16, 724 (1993)

[11] Cardellina JH, J. Liq. Chromatogr., 14, 661 (1991)

[12] Hamilton RJ, Sewell PA, "Introduction to High Performance Liquid Chromatography," 2nd Ed., Chapman and Hall, NY, U.S.A., 29 (1982)

[13] Kopycki WJ, Elsohly HN, Mc-Chesney JD, J. Liq. Chromatogr., 17, 2569 (1994)