The impact of packaging materials on the antioxidant phytochemical stability of aqueous infusions of green tea (Camellia sinensis) and yaupon holly (Ilex vomitoria) during cold storage.

Ready to drink (RTD) teas are a growing segment in the beverage category, brought about by improvements in the flavor of these products and healthy market trends driven by consumers. The presented results evaluated the antioxidant phytochemical stability of RTD teas from aqueous infusions of traditional green tea (Camellia sinensis) and a botanical tea from yaupon holly (Ilex vomitoria) as influenced by packaging materials during cold storage. Two common packaging materials for RTD products are glass and polyethylene terephthalate (PET) and have been compared to a retortable pouch (RP), an emerging packaging material for various types of food since it is durable, inexpensive, lightweight, and easy to sterilize. Storage stability was then evaluated for each aqueous infusion prepared at 10 g/L at 90 °C for 10 min and evaluated at 3 °C in the absence of light over 12 weeks. Analyses included quantification and characterization of individual polyphenolics by high-performance liquid chromatography-photodiode array and liquid chromatography-electrospray ionization-mass spectrometry as well as changes in total antioxidant capacity. For green tea, concentrations of the three major flavan-3-ols, epigallocatechin gallate, epigallocatechin, and epicatechin gallate were better retained in glass bottles as compared to other packages over 12 weeks. In yaupon holly, chlorogenic acid and its isomers that were the predominant compounds were generally stable in each packaging material, and a 20.6-fold higher amount of saponin was found as compared to green tea, which caused higher stability of flavonol glycosides present in yaupon holly during storage. The antioxidant capacity of green tea was better retained in glass and PET versus RP, whereas no differences were again observed for yaupon holly. Results highlight the superiority of oxygen-impervious glass packaging, but viable alternatives may be utilizable for RTD teas with variable phytochemical compositions.

Flavonol-rich fractions of yaupon holly leaves (Ilex vomitoria, Aquifoliaceae) induce microRNA-146a and have anti-inflammatory and chemopreventive effects in intestinal myofibroblast CCD-18Co cells.

Polyphenolics extracted from yaupon holly (Ilex vomitoria, Aquifoliaceae) (YH) leaves were investigated in human colon cells for their chemopreventive and anti-inflammatory activities. An activity-guided fractionation allowed the selection of YH flavonol-rich fraction due to its preferential inhibition of HT-29 colon cancer viability over the normal CCD-18Co colon cells. Quercetin and kaempferol 3-rutinosides, main components identified in this fraction, protected CCD-18Co cells against reactive oxidative species (ROS) in part due to increased activity of antioxidant enzymes. In addition, up-regulation of microRNA-146a (miR-146a) known as a negative regulator of pro-inflammatory NF-κB activation was the underlying molecular mechanism that protected CCD-18Co from inflammation.

Nitrogen fertilizer and gender effects on the secondary metabolism of yaupon, a caffeine-containing North American holly.

Yaupon (Ilex vomitoria) is a caffeine-containing dioecious shrub native to the southeastern United States that was historically brewed into a stimulating beverage. We tested predictions of the carbon/nutrient balance (CNB) hypothesis by determining whether nitrogen availability and gender influence production of caffeine and related alkaloids as well as phenolic compounds in leaves of pot-grown yaupon plants fertilized with ammonium nitrate. The CNB hypothesis predicts that additional nitrogen should result in increased alkaloid concentrations and decreased phenolic concentrations. An extension of the CNB hypothesis to dioecious plants predicts that females have higher C/N ratios and therefore higher phenolic concentrations and lower alkaloid concentrations than male conspecifics. In our study, caffeine and total alkaloid concentrations were 5-10 times higher in fertilized than control plants but did not vary by gender. Nevertheless, an observed interaction between gender and fertilization suggests that females respond more to fertilization than males in caffeine production. In addition, fertilized plants not only contained higher concentrations of alkaloids and total nitrogen but also allocated a larger proportion of their nitrogen to alkaloid production than control plants. Total phenolic concentrations were higher in control females than control males as predicted by the CNB hypothesis, but did not vary by treatment nor were there differences by gender among fertilized plants. We also found high correlations between antioxidant capacity and both classes of phenolic compounds detected in our study (cinnamic acid derivatives and flavonoids) indicating that in addition to their putative defensive function against herbivores, phenolics protect yaupon from oxidative stress. Explanation of the inconsistencies between our data and predictions of the CNB hypothesis may benefit from a re-appraisal of the physiological mechanisms by which resource availability affects secondary metabolism as well as consideration of the selective pressures to which secondary metabolism responds.

An improved thin-layer chromatography/mass spectrometry coupling using a surface sampling probe electrospray ion trap system.

A combined surface sampling probe/electrospray emitter coupled with an ion trap mass spectrometer was used for the direct read out of unmodified reversed-phase C18 thin-layer chromatography (TLC) plates. The operation of the surface sampling electrospray ionization interface in positive and negative ionization modes was demonstrated through the direct analysis of TLC plates on which a commercial test mix comprised of four dye compounds viz., rhodamine B, fluorescein, naphthol blue black, and fast green FCF, and an extract of the caffeine-containing plant Ilex vomitoria, were spotted and developed. Acquisition of full-scan mass spectra and automated collection of MS/MS product ion spectra while scanning a development lane along the surface of a TLC plate demonstrated the advantages of using an ion trap in this combination. Details of the sampling system, benefits of analyzing a developed lane in both positive ion and negative ion modes, levels of detection while surface scanning, surface scan speed effects, and the utility of three-dimensional data display, are also discussed.