On‐line concentration and pressurized capillary electrochromatographic analysis of phytohormones in corn

In the experiment, a high-performance liquid chromatography and electrospray ionization-tandem mass spectrometry with selected reaction monitoring was used to simultaneously determine various classes of phytohormones, including indole-3-acetic acid, α-naphthaleneacetic acid, 2-chlorobenzoic acid, 4-chlorobenzoic acid, indole-3-butyric acid, gibberellic acid, 2,4-dichlorophenoxyacetic acid, 2-naphthoxyacetic acid, abscisic acid, 2,3,5-triiodobenzoic acid, uniconazole, paclobutrazol and 2,4-epibassinolide in rape tissues. The analyses were separated by an HPLC equipped with a reversed-phase column using a binary solvent system composed of methanol and water, both containing 0.1% of formic acid. The matrix effect was also considered and determined. The technology was applied to analyze rape tissues, including roots, stems, leaves, flowers, immature pods and rape seeds. The rape tissues were subjected to ultrasound-assisted extraction and purified by dispersive solid-phase extraction, and then transferred into the liquid chromatography system. The detection limit for each plant hormone was defined by the ratio of signal/background noise (S/N) of 3. The results showed perfect linearity (R(2) values of 0.9987-1.0000) and reproducibility of elution times (relative standard deviations, RSDs,<1%) and peak areas (RSDs,<7%) for all target compounds.

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Simultaneous determination of 13 phytohormones in oilseed rape tissues by liquid chromatography‐electrospray tandem mass spectrometry and the evaluation of the matrix effect

Fan

Wang

et al. 2011

“…The flavonoids extraction procedure was similar to the method described previously with some modification [39]. Briefly, 4 mL of cold methanol ( -208C) was added to the tube containing the corn sample (120 mg) and incubated at -208C for 30 min.…”

Section: Sample Preparationmentioning

confidence: 99%

“…In order to enhance the detection sensitivity and enable the technique to analyze low levels of flavonoids in corn samples, gradient elution mode was used as an online sample concentration method to improve the detection sensitivity by increasing the sample injection volume without adverse effect on the band broadening. In our recently published papers [32,39], pCEC methods with isocratic elution was applied to the separation of water-soluble vitamins and phytohormones in corn and solvent gradient zone sharpening effect and field-enhanced sample stacking as online concentration methods were combined to improve the detection sensitivities of the analytes.…”

Section: Introductionmentioning

confidence: 99%

Pressurized CEC with gradient elution for separation of flavonoids from corn

Wang

Li²,

Chen

2009

Self Cite

A pressurized CEC (pCEC) method with gradient elution was developed for the analysis of flavonoids in two genetically modified corns and five nongenetically modified corns. The pCEC separation of flavonoids was performed on a monolithic silica-ODS column with UV detection at 270 nm. The effects of the concentration of organic solvent in the mobile phase, pH of the electrolyte buffer, applied voltage, and the gradient elution conditions were evaluated systematically. Gradient elution mode was used as an online concentration method to improve the detection sensitivity of flavonoids. Compared to the conventional injection (0.2 muL), there were 9- to 20-fold improvements in the detection sensitivity at the injection volume of 5 muL without adverse effect on the band broadening and the separation resolution. The LODs for the six flavonoids were in the range of 6.5-46 ng/mL under the gradient elution conditions. The developed gradient pCEC method was applied to evaluate flavonoids in various types of corns. The contents of flavonoids in genetically modified corns and nongenetically modified corns were compared.

“…Though monolithic columns are regarded as the promising separation channel, the related research is still in initial stage, there are no mature commercial columns yet. So far, there are few papers for plant hormones analysis based on CEC or pCEC [23,24]. In work by Yin and Liu [23], the open-tubular capillary column using polydopamine coating was prepared, then four Aux (including indole-3-butyric acid, 2,4-dichlorophenoxyacetic acid, IAA and phenoxyacetic acid) were chosen to evaluate and validate the performance of the above column in CEC system.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous analysis of endogenetic and ectogenic plant hormones by pressurized capillary electrochromatography

Zhang

Chen

et al. 2010

Pressurized CEC (pCEC) with UV detection was developed for the simultaneous analysis of endogenetic and ectogenic plant hormones. Effects of some crucial factors including mobile phase, buffer concentration, pH value and applied voltage on pCEC separation were investigated, respectively. Under the optimum conditions, namely a packed capillary column (3 microm Hypersil C(18), 45 cm total length with 100 microm id x 20 cm effective length) as the separation channel; ACN: 10 mmol/L phosphate (50:50, v/v, %) as the mobile phase, pH 4.0; pump flow rate 0.035 mL/min; separation voltage -10 kV; UV wavelength 222 nm, a complete separation of five hormones was accomplished. The pCEC assay presented good linearity (0.8-100 microg/mL), with LOD of 0.2 microg/mL and R in range of 0.998-0.999. Meanwhile, a lack-of-fit test was performed to evaluate these linear models. Results showed that no significant lack-of-fit was observed and these models were appropriate (alpha=0.05).