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首页> 外文期刊>International Journal of Electrochemical Science >Rapid and Ultrasensitive Method for Determination of Phytochelatin2 using High Performance Liquid Chromatography with Electrochemical Detection
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Rapid and Ultrasensitive Method for Determination of Phytochelatin2 using High Performance Liquid Chromatography with Electrochemical Detection

机译：高效液相色谱-电化学检测法快速，超灵敏地测定植物螯合素 2

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Plants respond to heavy metal toxicity in a variety of different ways including synthesis ofphytochelatins. The synthesis of phytochelatins is catalyzed by -Glu-Cys dipeptidyl transpeptidasenamed as phytochelatin synthase (PCS). The main aim of this study was to optimize high performanceliquid chromatography coupled with electrochemical detector for determination of phytochelatin2. Theoptimized procedure was subsequently used for determination of the mentioned molecules with specialattention aimed at the possibility to determine PC2 after activation of PCS in the tobacco BY-2 cellstreated with different concentrations of cadmium(II) ions. The optimized conditions were as follows.Both the detector and the column were thermostated at 30 C. Mobile phase consisted of A:trifluoroacetic acid (80 mM) and B: 100% methanol. Compounds were eluted by the following linearincreasing gradient: 0-1 min (3 % of B), 12 min (20 % of B), 125 min (98 % of B), 150 min-1 (98 % of B). Flow rate of the mobile phase was 1 ml min . Detection was carried out at appliedpotential 900 mV postponed on four electrodes. Time of one analysis was 15 minutes. The estimateddetection limit for PC2 was 17 nM. In addition, the recoveries were from 93 to 98 %. Good precisionwas obtained with %C.V.s ranging from 5.3 to 7.5 % in the intra-assay. The inter-assay %C.V.s rangedfrom 8.9 to 10.2 %. Overall recoveries of were from 102 to 106 % (n = 30). Accuracy (%Bias) wasabout 5 %. Further, the attention was aimed at determination of PC2 in BY-2 tobacco cells treatedInt. J. Electrochem. Sci., Vol. 6, 2011 1368with cadmium(II) ions (0, 5, 10, 25, 50 and 100 M). It clearly follow from the results obtained thatthe content of PC2 enhanced with increasing concentration of the substrate and with the appliedconcentration of cadmium(II) ions.
机译：植物以多种不同方式对重金属毒性做出反应，包括植物螯合素的合成。植物螯合素的合成被称为植物螯合素合酶（PCS）的-Glu-Cys二肽基转肽酶催化。这项研究的主要目的是优化高效液相色谱法和电化学检测器，用于测定植物螯合素2。随后将优化程序用于确定上述分子的检测，并特别注意在用不同浓度的镉（II）离子处理的烟草BY-2细胞中PCS活化后确定PC2的可能性。优化条件如下：检测器和色谱柱均在30℃恒温。流动相由A：三氟乙酸（80 mM）和B：100％甲醇组成。通过以下线性增加的梯度洗脱化合物：0-1分钟（B的3％），12分钟（B的20％），125分钟（B的98％），150 min-1（B的98％）。流动相的流速为1 ml min。在推迟在四个电极上施加的900 mV电位下进行检测。一次分析的时间为15分钟。 PC2的估计检测限为17 nM。此外，回收率从93％到98％。批内分析的％C.V.s在5.3％至7.5％范围内获得了良好的精度。批间测定的％C.V.s为8.9-10.2％。的总回收率为102％至106％（n = 30）。准确度（％Bias）约为5％。此外，注意力集中在确定BY-2烟草处理过的Int细胞中的PC2。 J.电化学。科学，卷6，2011 1368，含镉离子（0、5、10、25、50和100 M）。从获得的结果可以清楚地看出，PC2的含量随着底物浓度的增加和镉（II）离子浓度的增加而增加。

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