柱前衍生反相高效液相色谱法同时测定植物络合素(PCn)等巯基化合物

Translated title of the contribution: Analysis of Phytochelatins and Other Thiol-containing Compounds by RP-HPLC with Monobromobimane Pre-column Derivatization

孙琴, 叶志鸿, 王晓蓉, 黄铭洪

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Abstract

介绍一种荧光试剂monobromobimane柱前衍生,反相高效液相色谱-荧光检测测定标准样品和植物组织中植物络合素(PCn)等巯基化合物的方法。结果表明,采用乙腈和0.1%三氟乙酸组成的二元梯度洗脱程序,标准混合液中PCn等5种巯基化合物15min内得到很好的分离。以谷胱甘肽为代表考察方法的精密度,在1.25~160ng/μL浓度范围内与峰高有较好的线性关系,相关系数r2=0.9999。谷胱甘肽的回收率平均为97.64%,40d内的变异系数平均为2.21%。方法具有快速、灵敏、稳定性好、易于实施、同时测定PCn等多种巯基化合物等优点。通过该方法的应用,首次揭示Cd超积累植物东南景天对Cd的超量积累和解毒与PCn无关。

In this paper phytochelatins (PCn) and other thiol-containing compounds in the mixed standard solution and plant tissues were analyzed by RP-HPLC separation and fluorescence detection with monobromobimane pre-column derivatization. The results showed that by using ACN-0.1 % TFA binary gradient elution, PCn and other thiol-containing compounds in the mixed standard solutions were rapidly separated within 15 min. Glutathione was representatively selected to test the precision of this method. The calibration curve was linear in a wide range of 1.25~160 ng/μL (regression coefficient r~2=0.999 9). It was confirmed that this method was rapid, simple, highly sensitive, stable, and had a property of the simultaneous determination of PCn and other thiol-containing compounds. This method has been applied to determine PCn and other thiol-containing compounds in a Cd hyperaccumulator Sedum alfredii in response to Cd. It was found that Cd hyperaccumulation and detoxification in this plant is not based on PCn synthesis, since no PCn was detected in any tissue at any Cd treatment.
Translated title of the contributionAnalysis of Phytochelatins and Other Thiol-containing Compounds by RP-HPLC with Monobromobimane Pre-column Derivatization
Original languageChinese (Simplified)
Pages (from-to)304-310
Number of pages7
Journal南京大学学报(自然科学)
Volume41
Issue number3
Publication statusPublished - May 2005

User-Defined Keywords

  • 植物络合素
  • 巯基化合物
  • 反相高效液相色谱
  • 柱前衍生
  • monobromobimane
  • 东南景天
  • phytochelatins
  • thiol-containing compounds
  • RP-HPLC;pre-column derivatization
  • Sedum alfredii

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