The aim of this study was to test the performance of a HPLC method, designated for rapid quantification of chlorogenic acids (CGA) in green coffee extract (GCE). The precision statistics associated with the method were assessed using three independent laboratories with five samples analyzed in triplicate. Seven main CGA isomers (3-CQA, 5-CQA, 4-CQA, 5-FQA, 3,4-diCQA, 3,5-diCQA and 4,5-diCQA) were quantified. The concentration of total CGA in the samples varied from 32.24% to 52.65% w/w. The repeatability and reproducibility standard deviations for the determination of individual isomers varied, respectively, from 0.01 to 0.28 and 0.05-1.59. The repeatability and reproducibility standard deviations of the calculated total CGA, corresponding to the sum of the seven main CGA isomers, varied respectively, from 0.17 to 0.58 and 0.55-2.01. The fast HPLC method evaluated in this study was considered precise and appropriate for the determination of CGA in GCE.

An ultra-high-performance liquid chromatographic (UHPLC) separation was developed for six kava pyrones (methysticin, dihydromethysticin (DHM), kavain, dihydrokavain (DHK), desmethoxyyangonin (DMY), and yangonin), two unidentified components, and three Flavokavains (Flavokavain A, B, and C) in Piper methysticum (kava). The six major kavalactones and three flavokavains are completely separated (Rs > 1.5) within 15 min using a HSS T3 column and a mobile phase at 60 °C. All the peaks in the LC chromatogram of kava extract or standard solutions were structurally confirmed by LC-UV-MS/MS. The degradations of yangonin and flavokavains were observed among the method development. The degradation products were identified as cis-isomerization by MS/MS spectra. The isomerization was prevented or limited by sample preparation in a non-alcoholic solvent or with no water. The method uses the six kava pyrones and three flavokavains as external standards. The quantitative calibration curves are linear, covering a range of 0.5–75 μg/mL for the six kava pyrones and 0.05–7.5 μg/mL for the three flavokavains. The quantitation limits for methysticin, DHM, kavain, DHK, DMY, and yangonin are approximately 0.454, 0.480, 0.277, 0.686, 0.189, and 0.422 μg/mL. The limit of quantification (LOQs) of the three flavokavains are about 0.270, 0.062, and 0.303 μg/mL for flavokavain C (FKC), flavokavain A (FKA), and flavokavain B (FKB). The average recoveries at three different levels are 99.0–102.3% for kavalactones (KLs) and 98.1–102.9% for flavokavains (FKs). This study demonstrates that the method of analysis offers convenience and adequate sensitivity for determining methysticin, DHM, kavain, DHK, yangonin, DMY, FKA, FKB, and FKC in kava raw materials (root and CO2 extract) and finished products (dry-filled capsule and tablet).

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According to the Food and Agriculture Organization (FAO), protein demand is expected to increase globally by around 40% by 2030 as a response to the world’s population growth. Due to their clean label, vegan or vegetarian based applications, nutritional value, and cost-efficient properties, plant-based proteins have been widely studied. However, most of the alternatives currently found in the market have some challenges because of their poor solubility, emulsifying, gelling, and foaming attributes. Hemp seed protein has gained increasing attention due to its unique amino acids and fatty acids profiles. In this study, commercial HPC mixtures were adjusted to pH 2, 4, 6, 8, 10, and 12 followed by ultrasonication (US) for 5 min (5 s on: 5 s off) and incubated for an hour before neutralizing to pH 7. Following the treatments, the samples were analyzed for their hydrodynamic diameter, conductivity, zeta potential, polydispersity index, surface hydrophobicity, solubility, electrophoresis (SDS-PAGE), free sulfhydryl group, and optical characteristics. The samples treated with ultrasound at pH 8 and 10 significantly (p < 0.05) enhanced the solubility of the hemp seed protein by 12.12% and 19.05%, respectively. Similarly, the samples treated with ultrasonication and pH shifting at pH 6, 8, and 10 also significantly increased the amount of free sulfhydryl content (p < 0.05) to 41.6, 58.72, and 46.54 mmol/g from 32.8 mmol/g, respectively. This study shows that the application of ultrasonication and pH shifting is a promising alternative method to modify the functional properties of HPC and widen their applications in the food, cosmetics, and pharmaceutical industries.

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