Considerable inter- and intraspecific variation with respect to the quantity and composition of plant natural products exists. The processes that drive this variation remain largely unknown. Understanding which factors determine chemical diversity has the potential to shed light on plant defenses against herbivores and diseases and accelerate drug discovery. For centuries, Cinchona alkaloids were the primary treatment of malaria. Using Cinchona calisaya as a model, we generated genetic profiles of leaf samples from four plastid (trnL-F, matK, rps16, and ndhF) and one nuclear (ITS) DNA regions from twenty-two C. calisaya stands sampled in the Yungas region of Bolivia. Climatic and soil parameters were characterized and bark samples were analyzed for content of the four major alkaloids using HPLC-UV to explore the utility of evolutionary history (phylogeny) in determining variation within species of these compounds under natural conditions. A significant phylogenetic signal was found for the content of two out of four major Cinchona alkaloids (quinine and cinchonidine) and their total content. Climatic parameters, primarily driven by changing altitude, predicted 20.2% of the overall alkaloid variation, and geographical separation accounted for a further 9.7%. A clade of high alkaloid producing trees was identified that spanned a narrow range of altitudes, from 1,100 to 1,350 m. However, climate expressed by altitude was not a significant driver when accounting for phylogeny, suggesting that the chemical diversity is primarily driven by phylogeny. Comparisons of the relative effects of both environmental and genetic variability in determining plant chemical diversity have scarcely been performed at the genotypic level. In this study we demonstrate there is an essential need to do so if the extensive genotypic variation in plant biochemistry is to be fully understood.

The optimised HPLC method offers a simple and efficient quantification of the four major alkaloids. Copyright © 2017 John Wiley & Sons, Ltd.

In this study, we present a dissolution test system that allows for the testing of dissolution of nano- and micrometer size materials under highly controlled atmospheric composition (O2 and CO2), temperature, and pH. The system enables dissolution testing in physiological simulant fluids (here low-calcium Gamble’s solution and phagolysosomal simulant fluid) and derivation of the temporal dissolution rates and reactivity of test materials. The system was validated considering the initial dissolution rates and dissolution profiles using eight different materials (γ-Al2O3, TiO2 (NM-104 coated with Al2O3 and glycerin), ZnO (NM-110 and NM-113, uncoated; and NM-111 coated with triethoxycaprylsilane), SiO2 (NM-200—synthetic amorphous silica), CeO2 (NM-212), and bentonite (NM-600) showing high intra-laboratory repeatability and robustness across repeated testing (I, II, and III) in triplicate (replicate 1, 2, and 3) in low-calcium Gamble’s solution. A two-way repeated-measures ANOVA was used to determine the intra-laboratory repeatability in low-calcium Gamble’s solution, where Al2O3 (p = 0.5277), ZnO (NM-110, p = 0.6578), ZnO (NM-111, p = 0.0627), and ZnO (NM-113, p = 0.4210) showed statistical identical repeatability across repeated testing (I, II, and III). The dissolution of the materials was also tested in phagolysosomal simulant fluid to demonstrate the applicability of the ATempH SBR system in other physiological fluids. We further show the uncertainty levels at which dissolution can be determined using the ATempH SBR system.

An effective, robust, and inexpensive grinding device for the grinding of herb samples like bark and roots was developed by rebuilding a commercially available coffee grinder. The grinder was constructed to be able to provide various particle sizes, to be easy to clean, and to have a minimum of dead volume. The recovery of the sample when grinding as little as 50 mg of crude Cinchona bark was about 60%.Grinding is performed in seconds with no rise in temperature, and the grinder is easily disassembled to be cleaned. The influence of the particle size of the obtained powders on the recovery of analytes in extracts of Cinchona bark was investigated using HPLC.