凝胶净化-高效液相色谱法测定霉变甘蔗中3-硝基丙酸
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摘要
目的建立凝胶净化-高效液相色谱法测定霉变甘蔗中3-硝基丙酸(3-nitropropionic acid,3-NPA)的方法。方法样品利用乙腈涡旋震荡提取,提取液经凝胶渗透色谱净化后,用高效液相色谱法进行测定。色谱柱为Agilent Eclipse plus C_(18)柱(4.6 mm×150 mm,5μm),柱温40℃,进样体积10μL,流动相为A:0.02 mol/L的KH_2PO_4水溶液(以磷酸调pH 3.0),B:乙腈,等度洗脱。结果方法线性范围为0.5～20.0 mg/kg,相关系数为0.99989,方法检出限为0.05 mg/kg,定量限为0.15 mg/kg,不同浓度水平的加标回收率为91.7%～99.4%,精密度为7.23%～8.62%(n=6)。结论该方法稳定准确,适用于霉变甘蔗中3-硝基丙酸的检测。
OBJECTIVE To establish a method for determination of 3-nitropropionic acid(3-NPA) in moldy sugarcane by gel permeation chromatography-high performance liquid chromatography(GPC-HPLC) method. METHODS Samples were extracted by acetonitrile and cleaned up by GPC-HPLC. The sample extract was separated on an Agilent Eclipse plus C_(18) column(4.6 mm×150 mm,5 μm), keeping the column temperature at 40℃ with an injection volume of 10 μL. The condition of mobile phase was KH_2PO_4 aqueous solution(0.02 mol/L, pH 3.0 with phosphoric acid) as mobile phase A and acetonitrile as mobile phase B for isocratic elution. RESULTS There was a good linearity of the calibration curve for 3-NPA in the range of 0.5-20.0 mg/kg with the correlation coefficient of 0.99989. The detection limit of the method was 0.05 mg/kg, and the limit of quantification was 0.15 mg/kg. The average recovery at three spiked levels was in the range of 91.7%-99.4%,and the relative standard deviation was 7.23%-8.62%(n=6). CONCLUSION This method is stable and accurate, and is suitable for the detection of 3-nitropropionic acid in sugarcane.
OBJECTIVE To establish a method for determination of 3-nitropropionic acid(3-NPA) in moldy sugarcane by gel permeation chromatography-high performance liquid chromatography(GPC-HPLC) method. METHODS Samples were extracted by acetonitrile and cleaned up by GPC-HPLC. The sample extract was separated on an Agilent Eclipse plus C_(18) column(4.6 mm×150 mm,5 μm), keeping the column temperature at 40℃ with an injection volume of 10 μL. The condition of mobile phase was KH_2PO_4 aqueous solution(0.02 mol/L, pH 3.0 with phosphoric acid) as mobile phase A and acetonitrile as mobile phase B for isocratic elution. RESULTS There was a good linearity of the calibration curve for 3-NPA in the range of 0.5-20.0 mg/kg with the correlation coefficient of 0.99989. The detection limit of the method was 0.05 mg/kg, and the limit of quantification was 0.15 mg/kg. The average recovery at three spiked levels was in the range of 91.7%-99.4%,and the relative standard deviation was 7.23%-8.62%(n=6). CONCLUSION This method is stable and accurate, and is suitable for the detection of 3-nitropropionic acid in sugarcane.
引文
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[12] 李小林,刘敏,别伟,等.凝胶渗透色谱-高效液相色谱法测定食品中的8种抗氧化剂[J].现代仪器,2011,17(3):31-35.
[13] 刘兴玠,李秀芳,孙艳洁,等.变质甘蔗中毒的预防研究[J].卫生研究,1993,22(1):31-33.
[2] 张秋燕,白玉兰,高彩莲,等.变质甘蔗中毒临床随访和预防[J].中华儿童保健杂志,1998,6(2):92-93.
[3] 吴琳莉,李志龙,苏鹤,等.霉变甘蔗中毒16例临床报告[J].现代中医药,2007,2(27):22-23.
[4] 汪儆,雷祖玉,冯学勤,等.我国某些豆科植物中3-硝基丙酸的气相色谱和气相色谱-质谱研究[J].草地学报,1992,9(2):34.
[5] 江涛,张庆林,罗雪云,等.3-硝基丙酸的高效液相色谱分析[J].卫生研究,1999,28(5):300-302.
[6] 李兵,吴国华,刘伟,等.固相萃取-超高效液相色谱串联质谱法检测甘蔗中3-硝基丙酸的方法研究[J].中国食品卫生杂志,2012,24(2):127-132.
[7] 中华人民共和国卫生部.变质甘蔗食物中毒诊断标准及处理原则:WS/T 10—1996 [S].北京:中国标准出版社,1996.
[8] 胡文娟,王玉华,陈晓明,等.甘蔗及甘蔗汁中3-硝基丙酸的薄层色谱测定法[J].卫生研究,1988,17(5):39-42.
[9] 邵国建,姚建花,朱文涛.固相萃取-离子色谱法测定甘蔗中3-硝基丙酸[J].中国卫生检验杂志,2016,26(15):2154-2156.
[10] 刘勇,吴南,吴永宁,等.3-硝基丙酸的气相色谱-热能检定器测定法及其在大鼠体内的毒物代谢动力学研究[J].卫生研究,1993,22(1):35-38.
[11] 马娜,陈玲,熊飞.固相萃取技术及其研究进展[J].上海环境科学2002,21(3):181-184.
[12] 李小林,刘敏,别伟,等.凝胶渗透色谱-高效液相色谱法测定食品中的8种抗氧化剂[J].现代仪器,2011,17(3):31-35.
[13] 刘兴玠,李秀芳,孙艳洁,等.变质甘蔗中毒的预防研究[J].卫生研究,1993,22(1):31-33.