二维液相色谱测定绿豆芽中赤霉素、6-苄基腺嘌呤、4-氯苯氧乙酸和2,4-二氯苯氧乙酸
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摘要
在豆芽食品中对添加的植物生长调节剂的监控是十分重要的。在此基础上建立了绿豆芽抽提样品中赤霉素、6-苄基腺嘌呤、4-氯苯氧乙酸和2,4-二氯苯氧乙酸4种植物生长调节剂的二维液相色谱定量分析方法。实验中以ZB-10 C18(ODS-AP,10 mm×10 mm)为第一维色谱柱,V(甲酸)∶V(甲醇)∶V(水)=1∶10∶89溶液等度洗脱,实现绿豆芽样品的前处理净化;随后通过阀切换将分析物转移至第二维色谱柱(Supersil ODS 2. 5μm,4. 6 mm×150 mm),以甲醇-0. 1%甲酸水溶液为流动相梯度洗脱,并以紫外254 nm为检测波长实现上述4种植物生长调节剂的分离分析。该方法中,赤霉素、6-苄基腺嘌呤、4-氯苯氧乙酸和2,4-二氯苯氧乙酸的线性范围分别为0. 004~2、0. 000 1~0. 2、0. 005~2以及0. 008~2 mg/m L,检出限分别为1. 4、0. 03、1. 6、2. 4μg/m L;峰面积的RSD为2%~3%,回收率为95%~104%。实际样品的分析中,发现该绿豆芽样品中赤霉素和2,4-二氯苯氧乙酸的添加量为0. 018 mg/g和0. 006 mg/g,4-氯苯氧乙酸未检出。另外,还发现样品中由于存在干扰物而不能准确定量分析低浓度的6-苄基腺嘌呤。研究表明,该方法可以应用于绿豆芽的直接抽提样品,对其中的赤霉素、4-氯苯氧乙酸和2,4-二氯苯氧乙酸能准确进行定量分析,不需进一步的样品前处理。
A method for simultaneous determination of gibberellin acid,6-benzyladenine,4-chlorophenoxyacetic acid and 2,4-dichlorophenoxyacetic acid in mungbean sprouts was established by 2 D-LC.The separation was first performed on the first-dimension column ZB-10 C18( ODS-AP,10 mm×10 mm).The analysts were trapped on this column for purification and the mobile phase used here was V( formic acid) ∶ V( methanol) ∶ V( water) = 1 ∶ 10 ∶ 89 in isocratic elution mode. Then,the trapped analysts were transferred to the second-dimension column( Supersil ODS 2. 5 μm,4. 6 mm×150 mm) using valve-switching technique,methanol and 0. 1% formic acid water solutions were used as mobile phase in gradient elution mode.Thus,they could be determined at254 nm with a UV detector.The results showed that the linear ranges of gibberellin acid,6-benzyladenine,4-chlorophenoxyacetic acid and 2,4-dichlorophenoxyacetic acid were 0. 004 ~ 2,0. 000 1 ~ 0. 2,0. 005 ~ 2 and 0. 008 ~ 2 mg/m L,respectively; and their detection limits were 1. 4,0. 03,1. 6 and 2. 4 μg/m L,respectively.The RSD values were 2% ~ 3% and the recoveries ranged from95% ~ 104%.When those plant hormone residues were purified by the first-dimension column,they could be successfully detected in mungbean sprouts at the level of 0. 018 mg/g for gibberellin acid and 0. 006 mg/g for 2,4-dichlorophenoxyacetic acid; It was also found that 4-chlorophenoxyacetic acid was not detectable and 6-benzyladenine could not be accurately determined in these extracts.The method is accurate and reliable,and can be applied directly to the determination of gibberellins acid,4-chlorophenoxyaceti acid,and 2,4-dichlorophenoxyacetic acid from the extracts of mungbean sprouts without further sample preparation,but not for 6-benzyladenine.
A method for simultaneous determination of gibberellin acid,6-benzyladenine,4-chlorophenoxyacetic acid and 2,4-dichlorophenoxyacetic acid in mungbean sprouts was established by 2 D-LC.The separation was first performed on the first-dimension column ZB-10 C18( ODS-AP,10 mm×10 mm).The analysts were trapped on this column for purification and the mobile phase used here was V( formic acid) ∶ V( methanol) ∶ V( water) = 1 ∶ 10 ∶ 89 in isocratic elution mode. Then,the trapped analysts were transferred to the second-dimension column( Supersil ODS 2. 5 μm,4. 6 mm×150 mm) using valve-switching technique,methanol and 0. 1% formic acid water solutions were used as mobile phase in gradient elution mode.Thus,they could be determined at254 nm with a UV detector.The results showed that the linear ranges of gibberellin acid,6-benzyladenine,4-chlorophenoxyacetic acid and 2,4-dichlorophenoxyacetic acid were 0. 004 ~ 2,0. 000 1 ~ 0. 2,0. 005 ~ 2 and 0. 008 ~ 2 mg/m L,respectively; and their detection limits were 1. 4,0. 03,1. 6 and 2. 4 μg/m L,respectively.The RSD values were 2% ~ 3% and the recoveries ranged from95% ~ 104%.When those plant hormone residues were purified by the first-dimension column,they could be successfully detected in mungbean sprouts at the level of 0. 018 mg/g for gibberellin acid and 0. 006 mg/g for 2,4-dichlorophenoxyacetic acid; It was also found that 4-chlorophenoxyacetic acid was not detectable and 6-benzyladenine could not be accurately determined in these extracts.The method is accurate and reliable,and can be applied directly to the determination of gibberellins acid,4-chlorophenoxyaceti acid,and 2,4-dichlorophenoxyacetic acid from the extracts of mungbean sprouts without further sample preparation,but not for 6-benzyladenine.
引文
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[8]吴平谷,谭莹,张晶,等.分级净化结合气相色谱-质谱联用法测定豆芽中10种植物生长调节剂[J].分析化学,2014,42(6):866-871.
[9]陈亚玲,江阳,李永新,等.高效液相色谱法同时测定瓜果中6种植物生长调节剂[J].现代预防医学,2016,43(5):891-895.
[10]张亚莲,柳菡,王岁楼,等.高效液相色谱-串联质谱法测定豆芽中8种药物残留[J].分析测试学报,2015,34(2):164-170.
[11]中华人民共和国卫生部.粮食和蔬菜中2,4-滴残留量的测定:GB/T 5009.175—2003[S].北京:中国标准出版社,2003:435-439.
[12]浙江省质量技术监督局.无公害豆芽第3部分:6-苄基腺嘌呤残留量和4-氯苯氧乙酸钠残留量的测定:DB33/T 625.3—2007[S].杭州:浙江省质量技术监督局,2007-01-07.
[2]KIM K G,PARK D W,KANG G R,et al. Simultaneous determination of plant growth regulator and pesticides in bean sprouts by liquid chromatography-tandem mass spectrometry[J].Food Chem.,2016,208(7):239-244.
[3]BHARADWAJ L,DHAMI K,SCHNEBERGER D,et al.Altered gene expression in human hepatoma Hep G2 cells exposed to low-level 2,4-dichlorophenoxyacetic acid and potassium nitrate[J].Toxicol.In Vitro,2005,19(5):603-619.
[4]TROUDI A,SAMET A M,ZEGHAL N.Hepatotoxicity induced by gibberellic acid in adult rats and their progeny[J].Exp.Toxicol.Pathol.,2010,62(6):637-642.
[5]GUIOCHON G,MARCHETTI N,MRIZIQ K,et al.Implementations of two-dimensional liquid chromatography[J].J.Chromatogr.A,2008,1 189(1):109-168.
[6]MINSHULL T C,JOBY C,DOCKRELL D H,et al.Analysis of histone post translational modifications in primary monocyte derived macrophages using reverse phase*reverse phase chromatography in conjunction with porous graphitic carbon stationary phase[J]. J. Chromatogr. A,2016,1 453:43-53.
[7]WANG H X,WANG B,ZHOU Y,et al.Rapid and sensitive screening and selective quantification of antibiotics in human urine by two-dimensional ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry[J]. Anal. Bioanal. Chem.,2014,406(30):8 049-8 058.
[8]吴平谷,谭莹,张晶,等.分级净化结合气相色谱-质谱联用法测定豆芽中10种植物生长调节剂[J].分析化学,2014,42(6):866-871.
[9]陈亚玲,江阳,李永新,等.高效液相色谱法同时测定瓜果中6种植物生长调节剂[J].现代预防医学,2016,43(5):891-895.
[10]张亚莲,柳菡,王岁楼,等.高效液相色谱-串联质谱法测定豆芽中8种药物残留[J].分析测试学报,2015,34(2):164-170.
[11]中华人民共和国卫生部.粮食和蔬菜中2,4-滴残留量的测定:GB/T 5009.175—2003[S].北京:中国标准出版社,2003:435-439.
[12]浙江省质量技术监督局.无公害豆芽第3部分:6-苄基腺嘌呤残留量和4-氯苯氧乙酸钠残留量的测定:DB33/T 625.3—2007[S].杭州:浙江省质量技术监督局,2007-01-07.