Sample preparation using an absorbent for removal of polyphenols and a solid-phase extraction (SPE) cartridge for cleanup followed by high-performance liquid chromatography (HPLC) has been investigated for the simultaneous determination of eight neonicotinoid insecticides (dinotefuran, nitenpyram, thiamethoxam, imidacloprid, clothianidin, imidaclothiz, acetamiprid, and thiacloprid). After tea samples were soaked with water and extracted with acetonitrile, sample extracts were treated with an appropriate amount of polyvinylpolypyrrolidone (PVPP) to effectively remove polyphenols. The treated extract was cleaned up with a Carb-PSA cartridge. Neonicotinoid insecticides were eluted with acetonitrile from the cartridge and dried. The extract was redissolved with methanol/water (1:9, v/v) and analyzed by conventional HPLC coupled with an ultraviolet detector. The recoveries of eight neonicotinoid insecticides in tea samples were 71.4-106.6% at 0.1-1.0 mg kg(-1) spiked levels. Relative standard deviations were <10% for all of the recovery tests. The established method was simple, effective, and accurate and could be used for monitoring neonicotinoid insecticides in tea.
The uptake, translocation, metabolism, and distribution behavior of glyphosate in nontarget tea plant were investigated. The negative effects appeared to grown tea saplings when the nutrient solution contained glyphosate above 200 mg L. Glyphosate was highest in the roots of the tea plant, where it was also metabolized to aminomethyl phosphonic acid (AMPA). The glyphosate and AMPA in the roots were transported through the xylem or phloem to the stems and leaves. The amount of AMPA in the entire tea plant was less than 6.0% of the amount of glyphosate. The glyphosate level in fresh tea shoots was less than that in mature leaves at each day. These results indicated that free glyphosate in the soil can be continuously absorbed by, metabolized in, and transported from the roots of the tea tree into edible leaves, and therefore, free glyphosate residues in the soil should be controlled to produce teas free of glyphosate.
We describe a novel analytical method for quantification of free amino acids in tea using variable mobile phase pH, elution gradient and column temperature of reversed-phase high-performance liquid chromatography (RP-HPLC). The study of mobile phase pH 5.7 was chosen to simultaneous quantification of 19 free amino acids in tea, while it improved maximum resolution of glutamine, histidine and theanine. Elution gradient was adapted for enhancing the solution of free amino acids, mainly because of adjustment of mobile phase A and B. The column temperature of 40 °C was conducive to separate free amino acids in tea. The limit of detection (LOD) and limit of quantitation (LOQ) of this method were in the range of 0.097-0.228 nmol/mL and 0.323-0.761 nmol/mL, respectively. The relative standard deviation of intraday and interday ranged in 0.099-1.909% and 3.231-7.025%, respectively, indicating that the method was reproducible and precise, while recovery ranged between 81.06-112.78%, showing that the method had an acceptable accuracy. This method was applied for the quantification of free amino acids in six types of tea. Multivariate analysis identified serine, glutamine, theanine and leucine as the most influencing factor for classify among analyzed sample.
The use of an in vitro cell suspension to study insecticide metabolism is a simpler strategy compared to using intact plants, especially for a difficult matrix such as tea. In this study, a sterile tea leaf callus was inoculated into B liquid media with 2,4-dichlorophenoxyacetic acid (2,4-D, 1.0 mg L) and Kinetin (KT, 0.1 mg L). After 3-4 subcultures (28 days each), a good cell suspension was established. Utilizing these cultures, the metabolic behaviors of six insecticides, including two organophosphates (dimethoate, omethoate) and four neonicotinoids (thiamethoxam, imidacloprid, acetamiprid, and imidaclothiz) were compared. The results showed that thiamethoxam, dimethoate, and omethoate were easily metabolized by tea cells, with degradation ratios after 75 days of 55.3%, 90.4%, and 100%, respectively. Seven metabolites of thiamethoxan and two metabolites of dimethoate were found in treated cell cultures using mass-spectrometry, compared to only two metabolites for thiamethoxam and one for dimethoate in treated intact plants.
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