Fibrous Roots of Cimicifuga Are at Risk of Hepatotoxicity

The cause of liver damage by using black cohosh preparation has been concerned but remains unclear. After a preliminary investigation, the black cohosh medicinal materials sold in the market were adulterated with Asian cohosh (Cimicifuga) without removing the fibrous roots. The safety of Cimicifuga rhizome and fibrous roots is unknown and has not been reported. Therefore, in this paper, the rhizome and fibrous roots of Cimicifuga dahurica (Turcz.) Maxim (C. dahurica) were completely separated, extracted with 70% ethanol, and freeze-dried to obtain crude rhizome extract (RC) and fibrous roots extract (FRC). UHPLC-Q-TOF-MS was used to identify 39 compounds in the rhizome and fibrous roots of Cimicifuga, mainly saponins and phenolic acids. In the L-02 cytotoxicity experiment, the IC₅₀ of fibrous roots (1.26 mg/mL) was slightly lower than that of rhizomes (1.417 mg/mL). In the 90-day sub-chronic toxicity study, the FRC group significantly increased the level of white blood cells, ALP, ALT, AST, BILI and CHOL (p < 0.05); large area of granular degeneration and balloon degeneration occurred in liver tissue; and the expression of p-NF-kB in the nucleus increased in a dose-dependent manner. Overall, Fibrous roots of Cimicifuga are at risk of hepatotoxicity and should be strictly controlled and removed during the processing.     

Optimization of ultrasonic‐assisted extraction of phenolic antioxidants from Malus baccata (Linn.) Borkh. using response surface methodology

In this study, the optimum extraction conditions for maximum recovery of the content of total phenolics (TPC) and total antioxidant abilities were analyzed for Malus baccata (Linn.) Borkh. using response surface methodology. The effects of ethanol percentage (X₁,%), ultrasonic power (X₂, W) and extraction temperature (X₃, °C) on the total phenolic content (Y₁) and antioxidant ability (Y₂) were evaluated. A second‐order polynomial model produced a satisfactory fitting of the experimental data with regard to total phenolic content (R² = 0.9942, P < 0.0001) and antioxidant ability (R² = 0.9966, P < 0.0001). The optimized conditions were ethanol concentration of 61.0%, ultrasonic power of 308.6 W, extraction temperature of 51.1°C for TPC and 60.5%, 311.4 W, 51.6°C for antioxidant ability, the predicted values agreed well with the experimental values. Results implied that the major phenolic compounds in obtained extracts as chlorogenic acid, quercetin‐3‐gal/glu, quercetin‐3‐xyl/ara, phloretin‐2‐xyloside, quercetin‐3‐ rhamnoside, and phloridzin.     

Optimization of extraction process and the antioxidant activity spectrum–effect relationship of Angelica dahurica

Despite the large number of studies indicating that Angelica dahurica has strong antioxidant capacity, there are no clear details on the specific antioxidant components involved. In this study, the chromatograms and antioxidant activity of A. dahurica were obtained by gas chromatography–mass spectrometry (GC–MS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and ferric ion reducing antioxidant power (FRAP) methods. The factors affecting free radical scavenging were investigated under different extraction conditions, on the basis of the single-factor experiment. The results showed that the optimum conditions for the DPPH method were ultrasonic extraction, using 80% methanol as extract the extraction solvent, a 20:1 (ml/g) ratio of liquid to material and an extraction time of 30 min. Furthermore, the spectrum–effect relationship between the GC–MS chromatograms and the antioxidant effect of A. dahurica was established to evaluate the antioxidant components of A. dahurica using multiple data analysis methods. Isoimperatorin and byakangelicol made the greatest contribution to scavenging DPPH free radicals and ferric-reducing antioxidant power. This result may provide the basis for developing new and effective products based on the antioxidant ingredients of A. dahurica.     

Optimization of the ultrasound‐assisted extraction of antioxidant phloridzin from Lithocarpus polystachyus Rehd. using response surface methodology

The purpose of this study was to optimize the extraction process of phloridzin from Lithocarpus polystachyus Rehd. leaves using response surface methodology and to determine the antioxidant capacity of the extract. A Box–Behnken design was used to analyze the effects of ethanol concentration, liquid–solid ratio, soak time and extraction time on the extraction yield of phloridzin. The content of phloridzin was determined by high‐performance liquid chromatography. To assess the antioxidant capacity of the extract, three in vitro test systems were used (1,1‐,diphenyl‐2‐picrylhydrazyl, hydroxyl radical scavenging test and reduction force). The optimal parameters obtained by response surface methodology were a volume fraction of ethanol of 64%, a liquid–solid ratio of 37:1, a soaking time of 35 h and a sonication time of 38 min. The proportion of the extraction of phloridzin from L. polystachyus under these industrial process conditions was 3.83%. According to the obtained results, response surface methodology could be suggested as an adequate model for optimizing the extraction process of phloridzin from L. polystachyus . Ultrasound extraction significantly increased the extraction rate of phloridzin, which could be used as an antioxidant in pharmaceutical and food products.     

中药升麻的化学成分II. 升麻酰胺的化学结构

自兴安升麻(Cimicifuga dahurica)根茎分得一个新的桂皮酰胺衍生物, 命名为升麻酰胺(cimicifugamide, 1)经解析其IR、MS、1^H和13^C NMR、1^H-1^H COSY、1^H-1^H NOESY、13^C-1^H COSY、13^C-1^HCOLOC及选择性远程DEPTN图谱, 确证其结构为N-(3'-甲氧基-4'-羟基苯乙基)-4-O-β-D-吡喃半乳糖-阿魏酰胺。     

Identification of Pulsatilla chinensis (Bge.) Regel and look-alike species by UHPLC-TOF-MS using multivariate statistical analysis

Pulsatillae Radix, the root of Pulsatilla chinensis (Bge.) Regel, is recorded in the Pharmacopoeia of the People's Republic of China and has been widely used for its pharmacological activities, such as anti-inflammatory, antioxidant, antibacterial, antitumor, and cardiovascular benefits. However, there are several look-alike species that can be marketed as Pulsatillae Radix. To distinguish P. chinensis (Bge.) Regel from its look-alikes, viz. Pulsatilla cernua (Thunb.) Bercht et Opiz., Pulsatilla dahurica (Fisch.) Spreng., Anemone tomeutosa (Maxim.) Pei., and Rhaponticum uniflorum (L.) DC, we used ultra high performance liquid chromatography with time-of-flight mass spectrometry combined with principal component analysis to compare their chemical compositions. Four ions, a (RT 8.98 min, m/z 1381.6671), b (RT 10.64 min, m/z 1219.6143), c (RT 11.52 min, m/z 1217.5978), and d (RT 13.6 min, m/z 749.4463), from P. chinensis (Bge.) Regel were identified as potential chemical markers to distinguish it from look-alike species using an unsupervised statistical model combined with orthogonal partial least-squares discriminant analysis. The results of this study provide an effective method for identifying and distinguishing P. chinensis (Bge.) Regel from similar plants.     

Optimization of Microwave-Assisted Extraction (MAE) for the Isolation of Antioxidants from Basil (Ocimum basilicum L.) by Response Surface Methodology (RSM)

Abstract

The recovery of antioxidants from basil (Ocimum basilicum L.) was modeled with the aid of response surface methodology (RSM) using microwave-assisted extraction (MAE). Face-centered central design (FCCD) was employed to optimize the MAE operational parameters including the extraction time (1 to 7?min), extraction temperature (30 to 120?°C), solid-to-solvent ratio (0.1 to 0.4), and solvent concentration (20 to 80% ethanol, v/v), and to obtain the best possible combinations of these parameters for a high antioxidant yield from basil. The total antioxidant capacity (TAC) was expressed in trolox (TR) equivalents per gram of dried sample (DS). Three of the operational parameters (temperature, extraction time and solvent concentration) were shown to have significant effect on the extraction efficiency of antioxidants in basil extracts (p?<?0.05). The solvent concentration was shown to be the most significant factor on antioxidant yield obtained by MAE. There was a close relationship between experimental and predicted values using the proposed method. This optimized MAE method shows an application potential for the efficient extraction of antioxidants from basil in the food and pharmaceutical industries.     

Separation of three phenolic high‐molecular‐weight compounds from the crude extract of Terminalia Chebula Retz. by ultrasound‐assisted extraction and high‐speed counter‐current chromatography

This study presents an efficient strategy for separation of three phenolic compounds with high molecular weight from the crude extract of Terminalia chebula Retz. by ultrasound‐assisted extraction and high‐speed counter‐current chromatography. The ultrasound‐assisted extraction conditions were optimized by response surface methodology and the results showed the target compounds could be well enriched under the optimized extraction conditions. Then the crude extract was directly separated by high‐speed counter‐current chromatography without any pretreatment using n‐hexane/ethyl acetate/methanol/water (1:7:0.5:3, v/v/v/v) as the solvent system. In 180 min, 13 mg of A, 18 mg of B, and 9 mg of C were obtained from 200 mg of crude sample. Their structures were identified as Chebulagic acid (A, 954 Da), Chebulinic acid (B, 956 Da), and Ellagic acid (C) by ¹H NMR spectroscopy.     

Optimization of microwave‐assisted extraction for anthocyanins,polyphenols, and antioxidants from raspberry (Rubus Coreanus Miq.) using response surface methodology

Anthocyanins (Acys), polyphenols, and antioxidants were extracted from raspberry (Rubus Coreanus Miq.) using a highly efficient microwave‐assisted extraction technique. Different solvents, including methanol, ethanol, and acetone, were tested. The colors of the extracts varied from light yellow to purple red or dark red. SEM and other nutrient analyses verified that ethanol was the most favorable medium for the microwave‐assisted extraction of raspberry due to its high output and low toxicity. Effects of process parameters, including microwave power, irradiation time, and solvent concentration, were investigated through response surface methodology. Canonical analysis estimated that the highest total Acys content, total polyphenols content, and antioxidant activity of raspberry were 17.93 mg cyanidin‐3‐O‐glucoside equivalents per gram dry weight, 38.57 mg gallic acid equivalents per gram dry weight, and 81.24%, respectively. The polyphenol compositions of raspberry extract were identified by HPLC with diode array detection, and nine kinds of polyphenols were identified and quantified, revealing that chlorogenic acid, syringic acid, and rutin are the major polyphenols contained in raspberry fruits. Compared with other fruits and vegetables, raspberry contains higher Acy and polyphenol contents with stronger antioxidant activity, suggesting that raspberry fruits are a good source of natural food colorants and antioxidants.     

Microwave-Assisted Extraction of Curcuma longa L. Oil: Optimization,Chemical Structure and Composition,Antioxidant Activity and Comparison with Conventional Soxhlet Extraction

Curcuma root (Curcuma longa L.) is a very important plant in gastronomy and medicine for its unique antiseptic, anti-inflammatory, antimicrobial and antioxidant properties. Conventional methods for the extraction of curcuma oil require long extraction times and high temperatures that can degrade the active substances. Therefore, the objectives of the present study were: (i) first, to optimize the extraction yield of curcuma oil by applying a Box-Behnken experimental design using surface response methodology to the microwave-assisted extraction (MAE) technique (the independent variables studied were reaction time (10–30 min), microwave power (150–200 W) and curcuma powder/ethanol ratio (1:5–1:20; w/v); and, (ii) second, to assess the total phenolic content (TPC) and their antioxidant activity of the oil (at the optimum conditions point) and compare with the conventional Soxhlet technique. The optimum conditions for the MAE were found to be 29.99 min, 160 W and 1:20 w/v to obtain an optimum yield of 10.32%. Interestingly, the oil extracted by microwave-assisted extraction showed higher TPC and better antioxidant properties than the oil extracted with conventional Soxhlet technique. Thus, it was demonstrated that the method applied for extraction influences the final properties of the extracted Curcuma longa L. oil.     

Determination of phenolic compounds by MALDI–TOF and essential oil composition by GC–MS during three development stages of Origanum majorana L.

This study aimed to investigate the effect of the maturation process of sweet marjoram (Origanum majorana L.) on essential oil composition, the phenolic profile of ethanolic extract and their antioxidant capacities. The essential oil composition was studied at three stages of maturity by GC–MS. Thirty compounds were detected representing 100% of the total essential oil. p‐Menth‐1‐en‐4‐ol was the major compound (37.15–76.94%) followed by cyclohexanol‐3,3,5 trimethyl (5.41–15.99%) and α‐terpineol (0.94–11.34%). During the maturation process, an accumulation of oxygenated monoterpenes was observed. The phenolic composition was studied using matrix‐assisted laser desorption/ionization time of flight. The analysis showed the presence of short flavonoid monomers at all stages of maturation. The antioxidant capacity of ethanolic extracts and essential oils was evaluated using the DPPH assay, iron chelating power and reducing power assay. The highest phenolic content and antioxidant capacity were found at flowering stage. These findings on essential oil composition, phenolic profile and antioxidant capacity of O. majorana at three different stages of development provide more information on how these secondary metabolites are accumulated.     

UHPLC-MS/MS phenolic profiling and in vitro antioxidant activities of Inula graveolens (L.) Desf

Inula graveolens (L.) Desf. is an annual aromatic herb which has various uses on alternative medicine in many region of the world. In this study, antioxidant activities of ethanol and water extracts of the plant leaves were determined by in vitro DPPH method and phenolic composition of the plant sample was determined by LC-MS/MS analysis. The results showed that chlorogenic acid, quinic acid, hyperoside, protocatechuic acid and quercetin were the major phenolic compounds among the 27 standard compounds. The significant antioxidant capacity of the plant might be related with the high abundance of phenolic compounds.     

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.     

Bioactive Compounds from Ephedra fragilis: Extraction Optimization,Chemical Characterization,Antioxidant and AntiGlycation Activities

Response surface methodology (RSM) with a Box–Behnken design (BBD) was used to optimize the extraction of bioactive compounds from Ephedra fragilis. The results suggested that extraction with 61.93% ethanol at 44.43 °C for 15.84 h was the best solution for this combination of variables. The crude ethanol extract (CEE) obtained under optimum extraction conditions was sequentially fractionated with solvents of increasing polarity. The content of total phenolic (TP) and total flavonoid (TF) as well as the antioxidant and antiglycation activities were measured. The phytochemical fingerprint profile of the fraction with the highest activity was characterized by using RP-HPLC. The ethyl acetate fraction (EAF) had the highest TP and TF contents and exhibited the most potent antioxidant and antiglycation activities. The Pearson correlation analysis results showed that TP and TF contents were highly significantly correlated with the antioxidant and antiglycation activities. Totally, six compounds were identified in the EAF of E. fragilis, including four phenolic acids and two flavonoids. Additionally, molecular docking analysis also showed the possible connection between identified bioactive compounds and their mechanisms of action. Our results suggest new evidence on the antioxidant and antiglycation activities of E. fragilis bioactive compounds that may be applied in the treatment and prevention of aging and glycation-associated complications.     

Simultaneous quantitative assessment of nine glycosides in tobacco by liquid chromatography–tandem mass spectrometry

A simple and efficient method combining ultrasound‐assisted extraction, the conditions of which were optimized by response surface methodology, with liquid chromatography and tandem mass spectrometry was established and validated for the absolute quantification of nine non‐volatile neutral glycosides originating from tobacco (Nicotiana tobaccum L.) leaves, comprising three phenolic glycosides, one benzanoid glycoside, and five sesquiterpene glycosides within three isomers, originating from tobacco leaves. Factors of extraction time, sample quantity, extraction solvent, liquid chromatographic conditions, and electrospray ionization parameters were carefully investigated to ensure the selectivity and sensitivity of the method. All calibration curves showed excellent coefficients of determination ranging from 0.9940 to 0.9996, within the range of tested concentrations. The limits of detection and quantification were 2.33–25.9 and 7.06–78.5 ng/mL, respectively. Satisfactory values of accuracy were between 80.1 to 107.9% among different sample matrixes. The relative standard deviations of intra‐ and inter‐day analysis were less than 13.7 and 13.0% respectively. The developed method was successfully applied in a pilot study to determine the amounts of the nine endogenous glycosides in real flue‐cured tobacco samples obtained from different habitats in China.     

Separation of three anthraquinone glycosides including two isomers by preparative high‐performance liquid chromatography and high‐speed countercurrent chromatography from Rheum tanguticum Maxim. ex Balf

Anthraquinone glycosides, such as chrysophanol 1‐O‐β‐d‐ glucoside, chrysophanol 8‐O‐β‐d‐ glucoside, and physion 8‐O‐β‐d‐ glucoside, are the accepted important active components of Rheum tanguticum Maxim. ex Balf. due to their pharmacological properties: antifungal, antimicrobial, cytotoxic, and antioxidant activities. However, an effective method for the separation of the above‐mentioned anthraquinone glycosides from this herb is not currently available. Especially, greater difficulty existed in the separation of the two isomers chrysophanol 1‐O‐β‐d‐ glucoside and chrysophanol 8‐O‐β‐d‐ glucoside. This study demonstrated an efficient strategy based on preparative high‐performance liquid chromatography and high‐speed countercurrent chromatography for the separation of the above‐mentioned anthraquinone glycosides from Rheum tanguticum Maxim.ex Balf.     

Potentilla erecta (L.) rhizomes as a source of phenolic acids

The aim of this study was to evaluate the content of major phenolic acids from Potentilla erecta rhiozomes. Water and ethanol-water mixture was used for extraction of these compounds. The extracts were also evaluated for the quantification of total phenolic content and the antioxidant capacity. The contents of phenolic acids and resulting antioxidant activities are dependent on the nature of extracting solvent due to the presence of different antioxidant compounds. Results showed that P. erecta rhiozomes contained high amount of gallic and p-HBA acids. The contents of chlorogenic and protocatechuic acids in the extracts of Potentilla species have not been reported yet. The results suggested that the extracts could be used as the active cosmetics ingredients and nutraceuticals.     

Optimization of Ultrasound Assisted Extraction of Bioactive Compounds from Apple Pomace

In the present work, the optimization of the extraction of antioxidant compounds from apple pomace using ultrasound technology as an environmentally friendly and intensification process was developed. Different sonication powers, extraction temperatures and extraction times were studied and their influence on extraction yield and characteristics of the extracted samples (total phenolic compounds, flavonoid content and antioxidant capacity) are presented. The elaborated experimental design and the analysis of Pareto and response surface diagrams allowed us to determine the optimal extraction conditions. The conditions that allow the maximum extraction of phenolic compounds were found at 20 min, 90 °C and 50% ultrasound amplitude. Nevertheless, at these conditions, the antioxidant capacity measured by DPPH decreased in the extracted samples.     

Preparation and quantification of the total phenolic products in Citrus fruit using solid‐phase extraction coupled with high‐performance liquid chromatography with diode array and UV detection

Citrus fruit is an important health‐promoting food that is rich in dietary phenolic metabolites. Traditional Chinese medicines, such as Zhishi and Zhiqiao, come from young and immature fruits of Citrus cultivars. The preparation of diversified bioactive phenolic products and establishment of the corresponding quality control methodology are challenging and necessary. In the current study, four types of solid‐phase extraction sorbents for the enrichment and clean‐up of the phenolic matrix were evaluated. A solid‐phase extraction column coated with Strata‐X was finally used in the procedure. Twenty phenolic compounds were selected to evaluate the extraction performances of the sorbents using high‐performance liquid chromatography analysis. Under the optimized conditions, good linearities were obtained with R² more than 0.9996 for all analytes with LODs of 0.04–1.012 μg/g. Intra‐ and interday relative standard deviation values were less than 3%, and the recovery was equal to or higher than 90.02%. Compared to non‐solid‐phase extraction process, the content of total phenolic products was elevated 35.55–68.48% with solid‐phase extraction. Finally, the developed and validated method was successfully applied to the discrimination of Zhishi samples from different species as well as Zhishi and Zhiqiao samples in different development stages.     

Direct from solid natural products to pure compounds in a single step: Coupling online extraction with high‐speed counter‐current chromatography

Extraction is the most important step in the purification of bioactive compounds from natural products. This study introduces a simple online extraction strategy coupled with high‐speed counter‐current chromatography for efficient extraction and purification of bioactive components from solid natural products. For online extraction strategy, 1.0 g of ground Mangnolia officinalis or Piper nigrum was loaded into a guard column, which was then positioned on the manual injection valve instead of the sample loop. Bioactive components were directly extracted by the mobile phase of high‐speed counter‐current chromatography, and then transferred into high‐speed counter‐current chromatography for purification. In addition, the compatibility of the developed methodology for direct purification of bioactive components from fresh M. officinalis was successfully demonstrated. Obviously, in comparison with traditional offline heat‐reflux extraction, online extraction avoided the instrument, time, solvent, and energy consumption, and purified two phenolic compounds (honokiol and magnolol) from M. officinalis and three alkaloids (piperyline, piperine, and piperanine) from P. nigrum with high extraction efficiency. The superiority of the developed methodology is to establish an easy, rapid, and efficient technique for the purification of a wide variety of bioactive components from solid natural products.