Development of an Efficient HPLC Fluorescence Detection Method for Brassinolide by Ultrasonic-Assisted Dispersive Liquid–Liquid Microextraction Coupled with Derivatization
Development of an Efficient HPLC Fluorescence Detection Method for Brassinolide by Ultrasonic-Assisted Dispersive Liquid–Liquid Microextraction Coupled with Derivatization
复制标题
DOI：
10.1007/s10337-014-2767-9
复制
发表时间：
2014-10
期刊：
影响因子：
1.7
通讯作者：
Tao Lv;Xian-En Zhao;Shuyun Zhu;Zhongyin Ji;Guang Chen;Zhiwei Sun;Cuihua Song;J. You;Y. Suo-Y.-S
中科院分区：
文献类型：
--
作者：
Tao Lv;Xian-En Zhao;Shuyun Zhu;Zhongyin Ji;Guang Chen;Zhiwei Sun;Cuihua Song;J. You;Y. Suo-Y.-S
建立了超声辅助分散液-液微萃取(UA-DLLME)-衍生化联用技术用于油菜素内酯(油菜素内酯，油菜素内酯)的高效液相色谱荧光检测。用9-菲轮硼酸作BL的标记剂。对UA-DLLME萃取剂和分散剂的种类和用量、pH值、超声时间等参数进行了优化。优化了9-菲硼酸用量、吡啶体积比、衍生化时间和温度等衍生工艺参数。在最佳条件下，在L−1溶液中，布洛芬的定量线性范围为50-1,000 ng，线性响应良好，相关系数为0.9996。L−1的检出限为8.0ng，定量限为25.0ng.保留时间和峰面积的RSD分别为0.68-0.97%和4.61-6.54%，日内和日间分别为1.32-1.94%和7.28-9.75%。准确度在82.3-125.1%范围内令人满意。保留时间和峰面积的相对标准偏差分别为0.82-1.79和3.95-8.53%。BL的富集度为189。回收率在82.0-108.6之间，基质效应在90.0-115.3%之间。该方法已用于拟南芥、胡萝卜和油菜中盐基含量的测定。与许多其他方法相比，叶的灵敏度要高得多。
A novel hyphenated technique based on ultrasonic-assisted dispersive liquid–liquid microextraction (UA-DLLME) coupled with derivatization has been established for the determination of brassinolide (BL, a representative of brassinosteroids) by HPLC fluorescence detection. 9-Phenanthreneboronic acid is used as labeling reagent of BL. UA-DLLME parameters containing type and volume of extraction and disperser solvent, pH and ultrasonication time are optimized. Derivatization parameters are optimized included amount of 9-phenanthreneboronic acid, volume ratio of pyridine, derivatization time and temperature. Under optimal conditions, quantitative linear range of BL is 50–1,000 ng L−1and excellent linear response is observed with correlation coefficient of 0.9996. Limit of detection and limit of quantification are calculated as 8.0 and 25.0 ng L−1, respectively. RSDs of retention time and peak area are in the range of 0.68–0.97 % and 4.61–6.54 % for intra-day precision, 1.32–1.94 % and 7.28–9.75 % for inter-day precision, respectively. Accuracy is satisfactory in the range of 82.3–125.1 %. RSDs’ values of repeatability are in the range of 0.82–1.79 and 3.95–8.53 % for retention time and peak area, respectively. Enrichment factor for BL is 189. The results of recovery and matrix effect are in the range of 82.0–108.6 and 90.0–115.3 %, respectively. The proposed method has been applied for the determination of BL inArabidopsis thaliana,Daucus carotaandBrassica campestrisL. leaves with much higher sensitivity than many other methods.