Saito Kazunori, Ishii Hiroshi, Nishida Fumiko, Saito Hiroshi, Abe Toshihiko, Toyota Yoshimasa
School of Marine Science and Technology, Tokai University, 3-20-1 Orido, Shimizu, 424-8610, Shizuoka, Japan.
Toxicon. 2002 Jan;40(1):97-101. doi: 10.1016/s0041-0101(01)00201-x.
Microcystins (MCs) were purified by DEAE and C(18) cartridge chromatography. Addition of EtOH to the eluents (20%) in DEAE chromatography gave higher resolution than no addition of EtOH. The chromatogram showed three peaks: MC-LR; MC-LY and MC-LF; MC-LW. MC-LR and MC-LW were obtained by one step chromatography with purity of 96 and 88%, respectively. The separation of MC-LF and MC-LW with DEAE chromatography was better than that with reversed-phase chromatography. MC-LY and MC-LF were separated with C(18) cartridge. On the chromatogram, there were three peaks consisting of MC-LY (81% purity), MC-LF (86%), and an unknown compound which was considered as a MC variant judging from the results in HPLC/PDA, FAB-MS, and 1H NMR analyses, but the structure could not be determined. It is concluded that the combination of DEAE and C(18) cartridge chromatography would be a practical approach for the purification of various MCs.
微囊藻毒素（MCs）通过DEAE和C(18)柱色谱法进行纯化。在DEAE色谱法中，向洗脱液中添加乙醇（20%）比不添加乙醇具有更高的分辨率。色谱图上显示出三个峰：MC-LR；MC-LY和MC-LF；MC-LW。通过一步色谱法分别获得了纯度为96%和88%的MC-LR和MC-LW。用DEAE色谱法分离MC-LF和MC-LW比用反相色谱法效果更好。MC-LY和MC-LF用C(18)柱进行分离。在色谱图上，有三个峰，分别为纯度81%的MC-LY、纯度86%的MC-LF以及一种未知化合物，根据高效液相色谱/光电二极管阵列检测（HPLC/PDA）、快原子轰击质谱（FAB-MS）和核磁共振氢谱（1H NMR）分析结果判断该未知化合物为一种微囊藻毒素变体，但其结构无法确定。得出结论，DEAE和C(18)柱色谱法相结合是纯化各种微囊藻毒素的一种实用方法。
