R. J. Reynolds Tobacco Company

Background/Objectives: Date seeds contain a wide range of biologically active constituents and have been associated with multiple health benefits. Our study aimed to evaluate the utility of biphasic aqueous methanol/chloroform extraction for separating diverse metabolites from powdered Malakabi Ajwa date seeds and to identify the most effective chromatographic and spectroscopic techniques for their comprehensive profiling and structural characterization. Methods: Biphasic extraction generated organic, aqueous, and insoluble phases. Proper chromatography-mass spectrometry and spectroscopy analytical tools were then utilized to profile the metabolites in the aqueous and organic phases. Results and Conclusions: In the organic phase, fatty acids and their methyl esters were identified using GC-MS and confirmed by 1H NMR, while the absence of an O–H band in the FT-IR spectrum, together with 1H NMR data, supported the natural esterification of these fatty acids. In the aqueous phase, UV-Vis spectroscopy and LC-MS revealed anthocyanin pigments, and LC-MS additionally detected four amino acids. HPLC was used to quantify and validate the sugar composition, and 1H 1D and 2D NMR spectra enabled the assignment of α- and β-glucopyranose, β-fructopyranose, and sucrose signals. 31P NMR indicated that orthophosphate monoesters were the predominant phosphorus-containing compounds in both organic and aqueous phases. Overall, our findings show how an integrated chromatographic and spectroscopic workflow provides a robust strategy for comprehensive metabolite profiling of natural products such as Malakabi Ajwa date seeds.

Tobacco heating products (THPs) have reduced emissions of toxicants compared with cigarette smoke, and as they expose user to lower levels than smoking, have for a role to play in tobacco harm reduction. One key concern of Public Health is that new tobacco and nicotine products should not be more addictive than cigarettes. To assess their abuse liability, we determined nicotine pharmacokinetics and subjective effects of two THPs compared with conventional cigarettes and a nicotine replacement therapy (Nicotine inhaler). In a randomised, controlled, open-label, crossover study healthy adult smokers used a different study product in a 5 min ad libitum use session in each of four study periods. Product liking, overall intent to use again, urge for product and urge to smoke questionnaires were utilised to assess subjective effects. Nicotine uptake was greater for the cigarette (C max = 22.7 ng/mL) than for either THP (8.6 and 10.5 ng/mL) and the NRT (2.3 ng/mL). Median T max was significantly longer for the NRT (15.03 min) than for the tobacco products (4.05–6.03 min). Product liking and overall intent to use again was highest for the cigarette, and higher for the THPs than the NRT. Urge to smoke was reduced more by the cigarette than by the other three products. Urge to use the THPs was greater than the NRT. These findings suggest that the abuse liability of the THPs lies between that of subjects usual brand cigarettes and the NRT.

Ascorbic acid is a water-soluble vitamin common in food and dietary supplements. A usual problem with ascorbic acid analysis is the lack of stability of its samples and standard solutions owing to oxidation. A procedure to protect ascorbic acid from oxidation using mercaptoethanol is described in this study in connection with the comparison of three HPLC measuring methods. Two reversed-phase columns were evaluated for the separation. One technique uses UV detection, and two others use MS/MS detection. The methods were calibrated for quantitation on different ranges of concentrations. The LC-UV method covers the range 3.9 μg/ml to 500 μg/ml, one LC-MS/MS the range 80 ng/ml to 20 μg/ml, and the other 0.1 ng/ml to 20 μg/ml. As a proof of functionality all three methods were utilized for measuring vitamin C in energy drinks and chews (gummies). The sensitivity of LC-MS/MS methods was not necessary for the analysis of those samples, but the high sensitivity can be beneficial for other types of sample such as environmental or biological, where the levels of ascorbic acid are very low. The study showed that the formation of 2,3-diketogulonic acid is not a likely path for ascorbic acid oxidation following hydrolysis as reported in some studies.

In high-performance liquid chromatography, the dependence of retention factor k on volumetric fraction ϕ of organic phase is expressed by log k = F(ϕ) with F(ϕ) obtained by measuring log k at different ϕ values. From F(ϕ), a value kw is calculated by taking ϕ = 0. The equation log k = F(ϕ) is applied for predicting k, and kw is a descriptor of hydrophobic character of solutes and stationary phases. Calculated kw should not depend on the nature of organic component of mobile phase but extrapolation procedure leads to different kw for different organic components. The present study shows that the expression of F(ϕ) changes depending on the range of ϕ and the same function F(ϕ) cannot be used for the full range of ϕ from 0 to 1. Consequently, kw obtained by extrapolation of ϕ to zero is not correct because the expression of F(ϕ) was generated by fitting the data using ϕ with higher values. The present study shows the proper way to obtain the value of kw .

The retention behavior of six benzodiazepines (alprazolam, bromazepam, diazepam, flunitrazepam, medazepam, and nitrazepam) was studied using four different stationary phases, under reversed-phase mechanism in high-performance liquid chromatography. Four stationary phases were used for evaluating the retention of these compounds at fixed temperature. Functional dependences of the retention factor on the content of the organic modifier (methanol, or acetonitrile) in the composition of mobile phase were calculated. The extrapolated values of the retention factor for zero content of the organic modifier in mobile phase were higher for acetonitrile than for methanol for all studied compounds and for the four types of stationary phases.