Effective Isolation of Picrocrocin and Crocins from Saffron: From HPTLC to Working Standard Obtaining.

Saffron is widely cultivated and used as a spice. Recently published data on the chemical composition and pharmacological potential of saffron determine its use in pharmacy and medicine. The proposed high-performance thin-layer chromatography (HPTLC) method allows good separation of 11 analytes. The saffron quality (Iran, Ukraine, Spain, Morocco samples) assessment was based on the European Pharmacopoeia monograph and ISO 3632. The HPTLC method for the safranal, crocin, and picrocrocin quantification was proposed and validated. The crocins content in Ukrainian saffron was from 17.80% to 33.25%. Based on qualitative and quantitative assessment results, the saffron sample from Zaporizhzhia (Ukraine) had the highest compounds content and was chosen to obtain the working standards of picrocrocin and crocins (trans-4GG, trans-2G, trans-3Gg) by preparative chromatography. The compounds were isolated from lyophilized extract of saffron using a Symmetry Prep C18 column (300 × 19 mm × 7 µm), and identified by spectroscopic techniques (HPLC-DAD, UPLC-ESI-MS/MS). The purity of crocins and picrocrocin was more than 97%. A novel method proposed to obtain working standards is simple and reproducible for the routine analysis of saffron quality control.

Development of extraction technique and GC/FID method for the analysis of cannabinoids in Cannabis sativa L. spp. santicha (hemp).

INTRODUCTION: Nowadays, the interest in industrial Cannabis sativa L. herb has been increasing in the world. As a result, it is becoming one of the most studied plants due to its multifunctional benefits. OBJECTIVES: To the best of our knowledge, no study has been conducted so far to determine the impact of extraction methods and conditions on the extraction yields of CBD and CBG from the Cannabis sativa L. ssp. Santhica. Therefore, we aimed to investigate a simple and sensitive GC-FID method to determine CBD and CBG in hemp extract. METHODS: As regards sample preparation, three extraction techniques were compared, including maceration (ME), ultrasound-assisted extraction (UAE) and reflux-heat extraction (RHE), in order to obtain a high recovery of the CBD of interest from the plant material. The GC-FID method developed in this study represents a powerful tool for the extraction and analysis of non-psychoactive cannabinoids from hemp varieties to be used for the preparations of extracts with a high content of bioactive compounds for both pharmaceutical and nutraceutical applications. RESULTS: A simple extraction procedure for CBD and CBG from hemp was also optimized in this work, by using ultrasound assisted extraction method with 96% ethanol, material/solvent ratio 1:10 and extraction time 10 min at room temperature. CONCLUSION: The overall analytical method was fully validated in agreement with international guidelines. Therefore, proving a powerful and reliable tool for both the selection of hemp varieties with a high content of bioactive compounds and the quality control of its derivatives.

Analysis of content of phenolic acids in Lithuanian propolis using high-performance liquid chromatography technique.

UNLABELLED: The aim of the study was to analyze phenolic acids in Lithuanian propolis and to compare it with the composition of propolis in neighboring countries (Latvia and Poland) according to the predominant flora in the collecting places. The study was also aimed at the evaluation of the effect of the layer thickness (mm) of the harvested propolis on the quality of the raw material in determining the amount of phenolic acids. MATERIALS AND METHODS: The object of the study was propolis collected in Lithuania, Poland, and Latvia in late July of 2006 and 2007. The qualitative and quantitative analysis of phenolic acids was performed using the high-performance liquid chromatography technique (HPLC). RESULTS: The results of the study showed that the quantitative and qualitative composition of phenolic acids in propolis depended on the plants from which the bees in the area collected substances for the raw material of propolis. The predominant phenolic acids were determined to be ferulic and coumaric acids, and they may be among the main indicators of quality in the standardization of the raw material and preparations of propolis. CONCLUSION: We created an HPLC-based analysis method for the identification and quantification of phenolic acids in propolis. The variety of phenolic acids in propolis depends on the vegetation predominating in the harvesting area. Studies have shown that the highest amount of phenolic acids is observed in propolis harvested in areas characterized by the predominance of deciduous trees and meadows. Results have also shown that ferulic and coumaric acids are the predominant phenolic acids in propolis. The thickness of the layer of the collected propolis in the hive also influences its chemical composition.

Free radical-scavenging activities of Crataegus monogyna extracts.

UNLABELLED: The aim of this study was to investigate antiradical activity of aqueous and ethanolic hawthorn fruit extracts, their flavonoids, and flavonoid combinations. MATERIAL AND METHODS: Total amount of phenolic compounds and the constituents of flavonoids were determined using a high-performance liquid chromatography. The antioxidant activity of Crataegus monogyna extracts and flavonoids (chlorogenic acid, hyperoside, rutin, quercetin, vitexin-2O-rhamnoside, epicatechin, catechin, and procyanidin B(2)) quantitatively was determined using the method of spectrophotometry (diphenyl-1-picrylhydrazyl (DPPH.) radical scavenging assay and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid)(ABTS.+) radical cation decolorization assay). The level of tyrosine nitration inhibition was determined using a high-performance liquid chromatography. RESULTS: Ethanolic hawthorn fruit extract contained 182+/-4 mg/100 mL phenolic compounds, i.e. threefold more, as compared to aqueous extract. The antioxidant activity according to DPPH. reduction in the ethanolic extracts was higher 2.3 times (P<0.05). The ABTS.+ technique showed that the effect of ethanolic extracts was by 2.5 times stronger than that of aqueous extracts. Tyrosine nitration inhibition test showed that the effect of ethanolic extracts was by 1.4 times stronger than that of aqueous extracts. The investigation of the antiradical activity of the active constituents in aqueous and ethanolic extracts revealed that epicatechin and catechin contribute to radical-scavenging properties more than other components. Procyanidin B(2) only insignificantly influenced the antiradical activity of the extracts. CONCLUSION: Both aqueous and ethanolic hawthorn extracts had antiradical activity, but ethanolic extract had stronger free radical-scavenging properties, compared to the aqueous extract. The antioxidant activity of the studied preparations was mostly conditioned by epicatechin and catechin. The individual constituents of both extracts had weaker free radical-scavenging properties than the combination of these substances did.

The Influence of Different Oregano Species on the Antioxidant Activity Determined Using HPLC Postcolumn DPPH Method and Anticancer Activity of Carvacrol and Rosmarinic Acid.

The aim of this study was to evaluate concentration-dependent antioxidant and anticancer activities of CA and RA in ethanol extracts of three different Oregano species (Origanum onites L., Origanum vulgare L., and Origanum vulgare ssp. hirtum). The study revealed the highest RA antioxidant activity in O. vulgare ssp. hirtum (9550 ± 95 mmol/g) and the lowest in O. vulgare L. (2605 ± 52 mmol/g) (p < 0.05). The highest CA amount was present in O. onites L., which was 1.8 and 4.7 times higher (p < 0.05) than in O. vulgare ssp. hirtum and O. vulgare L., respectively. The anticancer activity was evaluated on human glioblastoma (U87) and triple-negative breast cancer (MDA-MB231) cell lines in vitro. RA anticancer activity was negligible. CA and the extracts were about 1.5-2 times more active against MDA-MB231 cell line (p < 0.05) compared to U87 cell line. The anticancer activities of three tested extracts were similar against U87 cell line (p > 0.05) but they had different activities against MDA-MB231 cell line.

Optimization of a CUPRAC-Based HPLC Postcolumn Assay and Its Applications for Ginkgo biloba L. Extracts.

The aim of the present work was to improve and validate the HPLC-CUPRAC postcolumn method for the evaluation of active antioxidant markers from the acetonic extracts of Ginkgo biloba leaves. Improvement of the HPLC online assay was performed by evaluating the suitable loop temperature, the reaction loop length, and the impact of flow rate. Separation of the analytes was performed by the HPLC method on an ACE C18 analytical column using a gradient elution program. The separated antioxidant markers in the extracts reacted with copper(II)-neocuproine (Cu(II)-Nc) reagent in the postcolumn reaction coil. The reagent was reduced by antioxidants to the copper(I)-neocuproine (Cu(I)-Nc) chelate with a maximum absorption at 450 nm. Validation experiments confirmed sufficient precision, sensitivity, and effectiveness of the corresponding method, which could be used for further evaluations of active antioxidant compounds in similar plant materials.