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  • High-internal-phase-emulsion polymeric monolith coupled with liquid chromatography-electrospray tandem mass spectrometry for enrichment and sensitive detection of trace cytokinins in plant samples.

High-internal-phase-emulsion polymeric monolith coupled with liquid chromatography-electrospray tandem mass spectrometry for enrichment and sensitive detection of trace cytokinins in plant samples.

Analytical and bioanalytical chemistry (2015-05-31)
Fuyou Du, Lin Sun, Xian Zhen, Honggang Nie, Yanjie Zheng, Guihua Ruan, Jianping Li
ABSTRACT

High-internal-phase-emulsion polymers (polyHIPEs) show great promise as solid-phase-extraction (SPE) materials because of the tremendous porosity and highly interconnected framework afforded by the high-internal-phase-emulsion (HIPE) technique. In this work, polyHIPE monolithic columns as novel SPE materials were prepared and applied to trace enrichment of cytokinins (CKs) from complex plant samples. The polyHIPE monoliths were synthesized via the in-situ polymerization of the continuous phase of a HIPE containing styrene (STY) and divinylbenzene (DVB) in a stainless column, and revealed highly efficient and selective enrichment ability for aromatic compounds. Under the optimized experimental conditions, a method using a monolithic polyHIPE column combined with liquid chromatography-electrospray tandem mass spectrometry (LC-MS-MS) was developed for the simultaneous extraction and sensitive determination of trans-zeatin (tZ), meta-topolin (mT), kinetin (K), and kinetin riboside (KR). The proposed method had good linearity, with correlation coefficients (R (2)) from 0.9957 to 0.9984, and low detection limits (LODs, S/N = 3) in the range 2.4-47 pg mL(-1) for the four CKs. The method was successfully applied to the determination of CKs in real plant samples, and obtained good recoveries ranging from 68.8 % to 103.0 % and relative standard deviations (RSDs) lower than 16 %.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Styrene, ReagentPlus®, contains 4-tert-butylcatechol as stabilizer, ≥99%
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Methanol, anhydrous, 99.8%
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Methanol, purification grade, 99.8%
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Kinetin, suitable for plant cell culture, crystalline
Sigma-Aldrich
Kinetin, suitable for plant cell culture, BioReagent, amorphous powder
Sigma-Aldrich
trans-Zeatin hydrochloride, suitable for plant cell culture, ≥97%
Sigma-Aldrich
Methanol, NMR reference standard
Sigma-Aldrich
Divinylbenzene, technical grade, 80%
Sigma-Aldrich
Divinylbenzene, technical grade, 55%
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Kinetin, ≥99.0% (HPLC)
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Gibberellin, 80% gibberellin A3 basis (TLC)
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Gibberellic acid, 90% gibberellin A3 basis (HPLC)
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Gibberellic acid, suitable for plant cell culture, BioReagent, ≥90% gibberellin A3 basis (of total gibberellins.)
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trans-Zeatin, BioReagent, suitable for plant cell culture, ≥97%
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Gibberellic acid potassium salt, suitable for plant cell culture, BioReagent, ~95%, ≥50% total GA3 basis
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Toluene, JIS special grade, ≥99.5%
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Methanol-12C, 99.95 atom % 12C
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Toluene, anhydrous, 99.8%
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Potassium persulfate, 99.99% trace metals basis
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Acetonitrile, electronic grade, 99.999% trace metals basis
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Isonicotinic acid, 99%
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