HPLC测定不同方法提取的山黧豆中β-ODAP  

Determination of β-N-oxalyl-α,β-diaminopropionic Acid in Lathyrus Sativus by HPLC with Different Extraction Methods

作　　者：赵兴秀[1] 何义国[1] 邓静[1] 方春玉[1] 孟延发[2] 

机构地区：[1]四川理工学院生物工程系,四川自贡643000 [2]四川大学生命科学学院,四川成都610064

出　　处：《食品研究与开发》2011年第9期145-148,共4页Food Research and Development

基　　金：四川省教育厅资助项目(08ZB043);自贡科技局项目(08);四川理工学院资助项目(2007ZR006)

摘　　要：利用2,4-二硝基氟苯(FDNB)为柱前衍生试剂,建立了测定山黧豆中毒素β-Ν-草酰基-α,β-二氨基丙酸(简称β-ODAP)及其无毒性的异构体α-N-草酰基α,β-二氨基丙酸(简称α-ODAP)的恒组分洗脱高效液相色谱方法。并优化了流动相的pH、乙腈含量以及柱温。得出最佳色谱条件为:C18,250 mm×46 mm,5μm;流动相∶乙腈/溶液A(17∶83,体积比;溶液A的配比是K2HPO4:二甲基甲酰胺:水为0.03 mol∶10 mL∶990 mL,并用冰醋酸调pH为5.5);流速1.0 mL/min;检测波长360 nm;柱温28℃。利用该方法比较了水提取法、30%乙醇提取法及0.2 mol/L高氯酸提取法的优缺点。结果发现:水提取法和0.2 mol/L高氯酸提取法效果相当,水提取法和30%乙醇提取法均耗时长,需要24 h,而耗时长会不可避免的导致β-ODAP向α-ODAP的转换,且30%乙醇提取法不如水提法和高氯酸提取法效果好。而0.2 mol/L高氯酸提取法耗时短,只需要在0℃～4℃下提取1 h。An isocratic high performance liquid chromatographic（HPLC） method was developed to analyze neurotoxinβ-Ν-oxalyl-α,β-diaminopropionic acid（β-ODAP） and its innoxious isomer,α-N-oxalyl-α,β-diaminopropionic acid（α-ODAP） in Lathyrus sativus by precolumn derivatization with 2,4-dinitrofluoro-benzene（FDNB）.The chromatographic conditions were optimized including pH of the buffer and the content of acetonitrile in mobile phase and column temperature.The optimized conditions were found such as column of C18（250 mm×46 mm,5 μm）,mobile phase consisted acetonitrile/liquor A（17∶83,volume ratio,Liquor A consisted of K2HPO4,dimethyl formamide and water in the proportion of 0.03 mol∶10 mL∶990 mL and adjusted to pH 5.5）,flow rate of 1.0 mL/min,detecting wavelength 360 nm and the column thermostated at 28 ℃.By the means the extraction method of water,30 % aqueous ethanol and 0.2 mol/L perchloric acid.As a result,the extraction methods of water and 0.2 mol/L perchloric acid effected better than that of aqueous ethanol.But the time of 0.2 mol/L perchloric acid extraction method was only needed 1 h under the condition of 0 ℃-4 ℃,much shorter than that of water and 30 % aqueous ethanol extraction methods,24 h.

关 键 词：山黧豆 β-N-草酰基-α β-二氨基丙酸 2 4-二硝基氟苯 高效液相色谱 

分 类 号：O657.7[理学—分析化学] TS207.3[理学—化学]