Biological Activity of Essential Oils of Four Juniper Species and Their Potential as Biopesticides.

The objective of this study was to assess the biological activity of essential oils (EOs) of four Juniperus species obtained via two different distillation methods and their potential as biopesticides. The studied factors were juniper species (Juniperus communis L., J. oxycedrus L., J. pygmaea C. Koch., and J. sibirica Burgsd), plant sex (male (M) and female (F)), and distillation method (hydrodistillation via a standard Clevenger apparatus (ClevA) and semi-commercial (SCom) steam distillation). The hypothesis was that the EO will have differential antioxidant, antimicrobial, and insecticidal activities as a function of plant species, plant sex, and distillation method. The two distillation methods resulted in similar EO composition within a given species. However, there were differences in the EO content (yield) due to the sex of the plant, and also differences in the proportions of some EO components. The concentration of α-pinene, ß-caryophyllene, δ-cadinene and δ-cadinol was dissimilar between the EO of M and F plants within all four species. Additionally, M and F plants of J. pygmaea, and J. sibirica had significantly different concentrations of sabinene within the respective species. The EOs obtained via ClevA extraction showed higher antioxidant capacity within a species compared with those from SCom extraction. All of the tested EOs had significant repellent and insecticidal activity against the two aphid species Rhopalosiphum padi (bird cherry-oat aphid) and Sitobion avenae (English grain aphid) at concentrations of the EO in the solution of 1%, 2.5%, and 5%. The tested EOs demonstrated moderate activity against selected pathogens Fusarium spp., Botrytis cinerea, Colletotrichum spp., Rhizoctonia solani and Cylindrocarpon pauciseptatum. The results demonstrate that the standard ClevA would provide comparable EO content and composition in comparison with SCom steam distillation; however, even slight differences in the EO composition may translate into differential bioactivity.

Use of water and ethanol extracts from wine grape seed pomace to prepare an antioxidant toothpaste.

BACKGROUND: Extracts of fresh wine grape seeds/skin or of grape pomace seeds were used to prepare antioxidant natural toothpastes. RESULTS: Ethanol extracted twice more polyphenols than water; ultrasound did not provide any improvement in the extraction. The addition of freeze-dried ethanol extracts of seeds or skin, at 2% and 10%, to the commercial toothpaste significantly increased the polyphenol content, both from white grape seeds and skin and from red grape seed pomace. The evaluation of time stability (shelf life) revealed a decrease, after 4 months, of 3.9% and 9.4% in total polyphenol content, in 5% and 10% water extracts, but not for ethanol extracts. 1,1-Diphenyl-2-picrilhydrazil1 antiradical activity was the highest in 10% of seed water extract toothpaste and, after 4 months, the activity was stable. CONCLUSION: Ethanol and water are efficient and safe solvents to create natural toothpaste with grape or pomace seed extract with antioxidant activity. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Optimization of antioxidant, anti-diabetic, and anti-inflammatory activities and ganoderic acid content of differentially dried Ganoderma lucidum using response surface methodology.

The dried Ganoderma lucidum (GL) has been widely used for its pharmacological properties and bioactive ganoderic acids (GAs). Herein, extraction procedures combining ultra-sonication and heating were optimized using response surface methodology based on four variables (antioxidant activity, anti-diabetic activity, total GAs content, and total polysaccharide content) and principal component analysis. The extraction of freeze-dried GL at temperatures between 64.2 and 70 °C for 1.2 h maximized the antioxidant activity and GA content, whereas the polysaccharide content and anti-diabetic activity were maximized by extraction between 66.8 and 70 °C for more than 2.8 h. Heat-dried GL extracted at 50 °C for 3 h provided the greatest anti-inflammatory activity against HaCaT cells by suppressing the response to inflammation related cytokines at mRNA levels. These results suggest that extraction conditions might be a limiting factor for target-oriented investigations, and optimized extraction methods may improve the potential effect and quality of harvested GL products.

Development of an aqueous ultrasound-assisted extraction process of bioactive compounds from beet leaves: a proposal for reducing losses and increasing biomass utilization.

BACKGROUND: Red beet plants are cultivated worldwide for the consumption of their roots, generating large amounts of unexploited by-products. In particular, beet leaves (BLs) represent about 50% of the whole plant and are usually discarded as waste. This constitutes not only an economic issue, since multiple resources invested in the production will be wasted, but also an environmental problem because of the pollution associated with their disposal. However, BLs comprise an important source of functional compounds (polyphenols and betalains) that could be recovered from the raw material, representing a sustainable solution for the underutilization of this by-product. This study proposes the recovery of polyphenols and betalains using an aqueous ultrasound-assisted extraction (UAE) process at different powers (35, 50, and 100 W) that was characterized and optimized. RESULTS: UAE significantly enhanced the recovery of bioactive compounds and shortened the time required for extraction in comparison with traditional macerations (35 < 50 < 100 W). During UAE, the temperature of the systems increased as a function of the power applied, favouring the recovery of these phytochemicals. Additionally, a Box-Behnken design and response surface methodology were employed to optimize UAE conditions (90 W ultrasound power, 1:20 solid:liquid ratio, 16 min extraction time), under which the yields were 14.9 mg g-1 (polyphenols), 949.1 µg g-1 (betaxanthins), and 562.2 µg g-1 (betacyanins), consistent with the values predicted by the models. CONCLUSION: This study enabled the development of a green-solvent UAE process that constitutes an effective post-harvest by-products strategy to minimize losses and increase biomass utilization through the recovery of bioactive compounds from BLs, promoting sustainability in the agri-food chain. © 2020 Society of Chemical Industry.

Environmentally Friendly Methods for Flavonoid Extraction from Plant Material: Impact of Their Operating Conditions on Yield and Antioxidant Properties.

The flavonoids are compounds synthesized by plants, and they have properties such as antioxidant, anticancer, anti-inflammatory, and antibacterial, among others. One of the most important bioactive properties of flavonoids is their antioxidant effect. Synthetic antioxidants have side toxic effects whilst natural antioxidants, such as flavonoids from natural sources, have relatively low toxicity. Therefore, it is important to incorporate flavonoids derived from natural sources in several products such as foods, cosmetics, and drugs. For this reason, there is currently a need to extract flavonoids from plant resources. In this review are described the most important parameters involved in the extraction of flavonoids by unconventional methods such as ultrasound, pressurized liquid extraction, mechanochemical, high hydrostatic pressure, supercritical fluid, negative pressure cavitation, intensification of vaporization by decompression to the vacuum, microwave, infrared, pulsed electric field, high-voltage electrical discharges, and enzyme-assisted extraction. There are no unified operation conditions to achieve high yields and purity. Notwithstanding, progress has been achieved in the development of more advanced and environmentally friendly methods of extraction. Although in literature are found important advances, a complete understanding of the extraction process in each of the unconventional techniques is needed to determine the thermodynamic and kinetic mechanisms that govern each of the techniques.

Influence of different extraction techniques on recovery, purity, antioxidant activities, and microstructure of flaxseed gum.

Four extraction techniques (that is, hot water extraction [HWE], alkaline-acidic extraction [AAE], ultrasound assisted extraction [UAE], and microwave assisted extraction [MAE]) were compared for flaxseed gum extraction and their influence on the yield, purity, structural characterization (monosaccharide composition, molecular weight distribution, and microstructure by transmission electron microscope), and antioxidant activity (in terms of scavenging ability of 2,2-diphenyl-1-picrylhydrazyl [DPPH], 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid [ABTS], and reducing power) was investigated. The HWE achieved higher yield (8.96%) than UAE (7.84%) followed by MAE (7.01%) and AAE (6.44%). Moreover, the four flaxseed gum (FSG) samples exhibited the identical monosaccharide composition, but slight difference was observed in the content, whereas the molecular weight ratios exhibited significant difference. All samples displayed concentration-dependent manner for all antioxidant assays. UAE-FSG showed significant higher scavenging ability on DPPH free radical, ABTS free radical, reducing power, and ß-carotene bleaching assay followed by HWE-FSG, MAE-FSG, and AAE-FSG. Overall results showed that UAE was favorable to the purity of FSG, whereas HWE was more advantageous to improve the extraction yield and facile as it requires no special equipment. PRACTICAL APPLICATION: Extraction methods directly affect the recovery and purity of the extracted compound. Thus, this study could help in selection of appropriate extraction method for FSG. The results suggested that FSG possesses potential healthcare application in food industry because of their nutrition composition and antioxidant activities, and thus, it can be used for formulation of functional food as a natural antioxidant agent.

Kinetic modeling for high voltage electrical discharge extraction based on discharge energy input.

In this work, a kinetic model for protein extraction from Camellia oleifera seed cake using high voltage electrical discharge extraction (HVED) was built with discharge energy inputs as primary variables. The results showed that both the equilibrium yields and the mass transfer coefficient of HVED were highly dependent on the HVED specific energy input per pulse (kJ/kg). After linear and nonlinear fitting with five different basic functions, the best model satisfied the power function relationship through optimizing the influence of specific energy input per pulse on the equilibrium yields and the mass transfer coefficients. Based on the observations, a predictive model that correlates the energy input, mass of raw material and kinetics of HVED extraction was proposed. The validity of the predictive model was verified, and the observed deviation was found to be less than 10%. This could provide a model basis for optimization of HVED at different processing capacities.

Biosurfactant trehalose lipid-enhanced ultrasound-assisted micellar extraction and determination of the main antioxidant compounds from functional plant tea.

Hydrosoluble trehalose lipid (a biosurfactant) was employed for the first time as a green extraction solution to extract the main antioxidant compounds (geniposidic acid, chlorogenic acid, caffeic acid, and rutin) from functional plant tea (Eucommia ulmoides leaves). Single-factor tests and response surface methodology were employed to optimize the extraction conditions for ultrasound-assisted micellar extraction combined with ultra-high-performance liquid chromatography in succession. A Box-Behnken design (three-level, three-factorial) was used to determine the effects of extraction solvent concentration (1-5 mg/mL), extraction solvent volume (5-15 mL), and extraction time (20-40 min) at a uniform ultrasonic power and temperature. In consequence, the best analyte extraction yields could be attained when the trehalose lipid solution concentration was prepared at 3 mg/mL, the trehalose lipid solution volume was 10 mL and the extraction time was set to 35 min. In addition, the recoveries of the antioxidants from Eucommia ulmoides leaves analyzed by this analytical method ranged from 98.2 to 102%. These results indicated that biosurfactant-enhanced ultrasound-assisted micellar extraction coupled with a simple ultra-high-performance liquid chromatography method could be effectively applied in the extraction and analysis of antioxidants from Eucommia ulmoides leaf samples.

Optimization of the extraction procedure for the determination of phenolic acids and flavonoids in the leaves of globe artichoke (Cynara cardunculus var. scolymus L.).

Globe artichoke (Cynara cardunculus var. scolymus L.) is not only used as a vegetable and ornamental plant, but also as important medicinal plant for the treatment of dyspeptic disorders. The European Pharmacopoeia describes a method for the quality assessment of dry artichoke leaves, which is time-consuming and requires huge amounts of organic solvents. In this study, an ultrasound-assisted extraction method was studied, which proved to be more efficient than the standard protocol of the European Pharmacopoeia, since it led to comparable results, was faster and easier to handle, and was more sustainable due to a reduced need for organic solvents.

Green solvents and Ultrasound-Assisted Extraction of bioactive orange (Citrus sinensis) peel compounds.

Byproducts such as orange peel have potential uses because of their bioactive compounds, which are important for their potential to reduce the risk factors of diseases caused by aging. The lack of effective techniques and the high levels of pollution produced by the conventional extraction of bioactive compounds using organic solvents have highlighted the need to enhance the 'green chemistry' trend. This study evaluates the use of ultrasound to extract bioactive compounds from orange peel. The antioxidant capacity, phenolic content, ascorbic acid, total carotenoids, and HPLC profile of phenolic compounds from orange peel extracts were obtained by a physicochemical evaluation. The results demonstrate that the optimal conditions for the ultrasound-assisted extraction of bioactive orange peel compounds were a power of 400 W, a time of 30 min, and 50% ethanol in water. These conditions were used to obtain a total carotenoid concentration of 0.63 mg ß-carotene/100 g, vitamin C concentration of 53.78 mg AA/100 g, phenolic concentration of 105.96 mg GAE/100 g, and antioxidant capacity of ORAC = 27.08 mM TE and TEAC = 3.97 mM TE. The major phenolic compound identified in all orange peel extracts was hesperidin, with a maximum concentration of 113.03 ± 0.08 mg/100 g.

Effects of acoustic power and pH on pectin-enriched extracts obtained from citrus by-products. Modelling of the extraction process.

BACKGROUND: The ultrasound-assisted extraction of pectic polysaccharides from orange by-products was investigated. Kinetics of mechanical agitation (0.2 × g) and acoustic (US1: 542 W L-1 and US2: 794 W L-1 ) extractions, were obtained and modelled at different pH values (1.5 and 2.0). All extractions were carried out at 25 °C, using citric acid as the extraction solvent. RESULTS: Higher pectic polysaccharides extraction yields were obtained with ultrasonic assistance, in comparison with the results obtained using mechanical agitation. Moreover, yield increases were significantly higher using the more acidic pH. Thus, at pH 1.5, pectin yield increased from ∼19%, obtained with agitation, to ∼47%, applying ultrasound; whereas, at pH 2.0, this increase was from ∼10%, with agitation, to ∼18%, applying ultrasound. A considerable decrease of the galacturonic acid proportion was observed on the extracts when ultrasound were applied for 60 min under pH 2.0. High methoxyl pectins were extracted at pH 1.5 whereas at pH 2.0, pectins exhibited a low methylation degree. Curves of acoustic and mechanical agitation extractions were properly represented by a second-order rate model (average mean relative error ≤ 7.4%). The extraction rate constant, initial extraction rate and maximum yield were determined for all experimental conditions. CONCLUSION: Overall, the results clearly indicated that the effect of ultrasound was highly dependent on the pH. Therefore, adequate acidic conditions must be applied in order to improve the efficiency of ultrasound on the pectin extraction process. © 2019 Society of Chemical Industry.

Partial propane extraction of aromatic compounds from dehydrated basils (Ocimum Lamiaceae).

BACKGROUND: Many conventional extraction methods for basils (Ocimum sp. Lamiaceae) produce only the extract as a usable product and leave the extracted herb as a waste product. We demonstrate partial extraction of chemically and morphologically diverse basil cultivars using propane at low temperature (20-27 °C) and pressure (950-1200 kPa) and evaluate the process for production of dual products, the extracted herb (raffinate) and the herb extract in terms of aromatic content and color. RESULTS: The extracts contained aromatic compounds that were characteristic of but not always identical in terms of relative abundance to the dehydrated herb. Extraction decreased total aromatics in the raffinate by 12-43% but the individual aromatic proportions remained essentially the same, preserving flavor characteristics of the raffinate. Color was mostly unchanged by the extraction process. CONCLUSION: Partial propane extraction resulted in two useful basil products (an extract and extraction raffinate). Aromatic extractability was tissue and cultivar dependent for basils. Therefore, partial extraction protocols should be optimized according to cultivar/plant tissue abundance to provide consistent aromatic intensity of these potential food products. © 2019 Society of Chemical Industry.

Combination of thermal pretreatment and alcohol-assisted aqueous processing for rapeseed oil extraction.

BACKGROUND: The alcohol-assisted aqueous extraction processing (AAEP) of oil has many advantages such as no need for demulsification and relative low cost compared with enzymatic aqueous extraction processing (EAEP). Three kinds of thermal pretreatments including dry-heating, wet-heating and soak-heating followed by the AAEP of rapeseed oil were investigated. RESULTS: Both soak-heating and wet-heating had a higher contribution rate to oil yield than dry-heating due to the enhancement of heat transfer rate owing to the high moisture content in the rapeseed cells. However, oil from soak-heated rapeseeds showed a much lower level on peroxide value (0.41 mmol kg-1 ) than that of wet-heated rapeseeds (5.23 mmol kg-1 ). In addition, transmission electron microscopy images illustrated that promoting effects of soak-heating and wet-heating on oil release were attributed to the coalescence of oil bodies. A relative low concentration of alcohol solution as an extraction medium, the highest oil recovery of 92.77% was achieved when ground rapeseeds (mean particle size: 21.23 µm) were treated with 45% (v/v) alcohol for 2 h at 70 °C and pH 9.0. Both the acid value and the peroxide value are lower than the commercial oil produced by extrusion and hexane extraction. Furthermore, the oil produced from AAEP also had higher content of tocopherols and lower content of trans-fatty acids than the commercial oil. CONCLUSION: AAEP of oil from soak-heated rapeseeds is a promising alternative to conventional oil extraction methods. © 2019 Society of Chemical Industry.

Establishment of a quantitative and qualitative analysis and isolation method for tetracyclic iridoids from Morinda lucida Bentham leaves.

A new high performance liquid chromatography (HPLC) method has been established for quantitative and qualitative analysis of three tetracyclic iridoids: ML-2-3 (1), molucidin (2), and ML-F52 (3), which are responsible for anti-trypanosomal and anti-leishmanial activities of Morinda lucida Bentham leaves. Separation of 1-3 from dried 80% aqueous (aq.) ethanol extract was achieved on a reversed-phase cholester column packed with cholesteryl-bonded silica using an acetonitrile-0.1% aq. formic acid mobile phase system. Ultraviolet-visible (UV-VIS) spectroscopy was employed for detection of compounds, and their contents were determined by measuring absorbance at 254 nm. Depending on the above system, several factors potentially affecting the concentration of tetracyclic iridoids were evaluated resulting in several variation on plant organs, seasonality, variation between individual trees, and branch positions within the trees. Moreover, we developed a simple, quick, and effective method for tetracyclic iridoid isolation from M. lucida leaves that consisted of extraction by sonication into 80% aq. ethanol, basic hydrolysis, acid neutralization, liquid-liquid extraction into an organic solvent, and reverse phase open column chromatography. Employing this method, we have succeeded to obtain 1 as a colorless crystal yielding of 0.23%, which was 28 times higher than that of previous isolation method. Setting up methodology in this paper may be important for future in vitro and in vivo studies of tetracyclic iridoids and moreover for their applications in new drug design and development.

Separation and analysis of flavonoid chemical constituents in flowers of Juglans regia L. by ultra-high-performance liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry.

The flowers of Juglans regia L. have demonstrated their medicinal value as a part of edible plants. In this study, an analytical methodology for identification of flavonoid chemical constituents in flowers of Juglans regia L. was established using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). Thirty-six flavonoid compounds were identified based on highly accurate mass measurements, and the mass spectrometric fragmentation pathways of eleven representative compounds were proposed, which was helpful for the identification of different types of flavonoids. The study has laid the foundation for the research and development of effective drugs from flowers of Juglans regia L.

Qualitative and quantitative analysis on the chemical constituents in Orthosiphon stamineus Benth. using ultra high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

A sensitive and efficient method was established and validated for qualitative and quantitative analysis on the chemical constituents in Orthosiphon stamineus Benth. (O. stamineus) using ultra high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. Based on the retention time and MS spectra, 61 compounds were detected by using ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry. 52 chemical structures in the O. stamineus extracts including 26 phenolic acids, 11 flavonoids, 6 diterpenoids, 4 fatty acids and 5 tanshinones were tentatively identified without the time-consuming process of isolation. Moreover, five chemical constituents (Danshensu, Caffeic acid, Rosmarinic acid, Sinensetin and Eupatorin) were quantified in three different batches of O. stamineus samples by the developed ultra high-performance liquid chromatography coupled with electrospray ionization triple-quadrupole mass spectrometry method in 10 min. The method validation of the five compounds was performed with acceptable linearity (R2, 0.9930-0.9997), precision (RSD, 1.87-10.36%), repeatability (RSD, 0.59-4.87%) and recovery (105.30-110.53%, RSD ≤ 13.90%).

New insights into phenol and polyphenol composition of Stevia rebaudiana leaves.

The diversity in phenols and polyphenols of stevia leaf has been simplified applying sequential fractionation techniques on its ethanol extract through ultrasound assisted maceration. Two of the fractions obtained by reverse-phase column chromatography resulted differently active in an extensive antioxidant and cytotoxic screening. Both fractions were chemically profiled by ultra-performance liquid chromatography (UHPLC) coupled with electrospray ionization (ESI) quadrupole/time-of-flight (QqTOF) mass spectrometry (MS). One of the fractions was composed mainly of chlorogenic acids and flavonol triglycosides, whereas the other was rich in flavonoids mono- and diglycosides and in their hydroxycinnamoyl derivatives. Among the fifty compounds identified, non-phenol metabolites, such as benzyl primeveroside and roseoside, as well as a lignan polyphenol (5'), are reported for the first time as constituents of the Stevia leaf.

Chemical profiles and quality evaluation of Buddleja officinalis flowers by HPLC-DAD and HPLC-Q-TOF-MS/MS.

The dried flowers and inflorescences of Buddleja officinalis Maxim are used as traditional medicines in China, and aqueous extracts of the flowers have also been used since ancient times as a yellow rice colorant at local festivals. In this study, HPLC-Q-TOF-MS/MS was used to determine the overall chemical composition of this medicine-food plant. A total of 54 compounds, including 23 flavonoids, 19 phenylethanoid glycosides, 7 alkaloids and 5 other compounds, were detected in the methanol extracts of the herb using this method. Among them, 35 compounds were found firstly in this herb. HPLC fingerprints were also developed, together with a method for the simultaneous quantification of 11 constituents that could be used for quality evaluation of B. officinalis. Fingerprint analysis, using 28 characteristic fingerprint peaks, was used to assess the similarities among 12 samples collected from different geographic areas and showed that the similarity was >0.900. Simultaneous quantification of 11 markers in B. officinalis was then performed to determine consistency of quality. Additionally, the total phenolic content and antioxidant capacity of extracts of the 12 samples of B. officinalis flowers were measured using spectroscopic methods. B. officinalis was found to have good antioxidant capacity and to be a potential natural antioxidant. The highest antioxidant capacity was found in the samples from Guizhou, Sichuan and Guangxi Province. Our results provide valuable information for further understanding and exploiting the herb.

A multidimensional analytical approach based on time-decoupled online comprehensive two-dimensional liquid chromatography coupled with ion mobility quadrupole time-of-flight mass spectrometry for the analysis of ginsenosides from white and red ginsengs.

Here, time-decoupled comprehensive two-dimensional ultra-high liquid chromatography (UHPLC) coupled with an ion mobility (IM)-high resolution mass spectrometer (HRMS) was established and used to analyze ginsenosides from the main roots of white ginseng (WG) and red ginseng (RG), which enabled the separation of complex samples in four dimensions (2D-LC, ion mobility, and mass spectrometry). The incompatibility of mobile phases, dilution effect, and long analysis time, which are the main shortcomings of traditional comprehensive 2D-LC methods, were largely avoided in this newly established 2D-UHPLC method. The orthogonality of this system was 55%, and the peak capacity was 4392. Under the optimized 2D-UHPLC-IM-MS method, 201 ginsenosides were detected from white and red ginseng samples. Among them, 10 pairs of co-eluting isobaric ginseng saponins that were not resolved by 2D-UHPLC-HRMS were further resolved using 2D-UHPLC-IM-MS. In addition, 24 ginsenoside references were analyzed by UHPLC-IM-MS to obtain their collision cross section (CCS) values and ion mobility characteristics. Finally, the established new method combined with multivariate statistical analysis was successfully applied to differentiate WG and RG, and 9 ginsenosides were found to be the potential biomarkers by S-Plot and the values of max fold change, which could be used for classifying WG and RG samples. Overall, the obtained results demonstrate the applicability and potential of the established time-decoupled online comprehensive 2D-UHPLC-IM-MS system, and it will be extended to the analysis of other targeted or untargeted compounds, especially co-eluting isomers in more herbal extracts.

Rapid discrimination between red and white ginseng based on unique mass-spectrometric features.

Red ginseng (RG) and white ginseng (WG), two processed products of Panax ginseng C. A. Meyer, are in high demand due to their unique features. In this study, some of these unique features were identified and confirmed as biomarkers of RG by using ultra-high-performance liquid chromatography-mass spectrometry, data mining, support vector machine, and artificial neural network. Principal component analysis showed clear separation between the RG and WG extracts, indicating the presence of potential discriminators. In addition, 20 features that are dominant in RG were found by data mining. Samples of Panax quinquefolium (PQ) and Panax notoginseng (PN), close relatives of Panax ginseng C.A.Meyer, were investigated and it was found that 17 features which were absent in PQ and PN samples, were present in RG and WG. Five of these markers were identified as nitrogen-containing compounds that have not been previously reported. Finally, we found that RG can be identified among different ginseng medicinal herbs including RG, WG, PQ, and PN samples, by loading four feature markers corresponding to nitrogen-containing compounds into a discriminating model, based on a support vector machine or an artificial neural network. Thus, this study provides an efficient tool to identify RG during pharmacological research.