Anthraquinone Content Research Articles

Influence of the drying method on the bioactive compounds and pharmacological activities of rhubarb.

Raw rhubarb samples that have been subjected to different drying procedures will have different therapeutic effects, possibly due to processing-induced variations in the chemical composition. In the present work, the fresh materials were processed by smoking, sun-drying, shade-drying and oven-drying at low, moderate and high temperatures. To facilitate the selection of a suitable drying method for rhubarb, the quality of rhubarb processed under various drying conditions was evaluated based on the simultaneous determination of multiple bioactive constituents in combination with bioactivity assays. The total concentrations of 12 compounds in smoked rhubarb were higher than the concentrations of the same components in raw rhubarb and rhubarb products processed using other drying techniques. Smoked rhubarb was found to substantially inhibit Na+ /K+ -ATPase and thrombin. In addition, higher content of anthraquinones led to higher Na+ /K+ -ATPase inhibitory activities, and higher gallic acid content increased the antithrombin capacity. The results confirmed that post-harvest fresh plant materials, especially roots, were still physiologically active organs that could undergo series of anti-dehydration mechanisms, including the production of related secondary metabolites during the early stages of dehydration. Therefore, the proper design of drying processes could enhance the quality of rhubarb as well as other similar medicinal plants. © 2018 Society of Chemical Industry.

Antibacterial activity of anthraquinone from cassia seed on spiced pig head

[Objective] To optimize the extraction of anthraquinone from cassia seed by ultrasonic extraction and its antibacterial activity. [Method] The influences of different extraction time and ethanol concentration, on anthraquinone content were evaluated by a single factor experiment. And anthraquinone content was determined by ultraviolet spectrophotometry. The bacteriostasis of anthraquinone on spiced pig head’s common putrefying bacteria: Staphylococcus, Serratieae, Bacillus, Proteus and the minimal inhibitory concentration (MIC) were studied by oxford plate assay system. [Result] The best extraction time was 30 minutes and the best ethanol concentration was 80%. The antibacterial activity of the cassia seed anthraquinone on Staphylococcus Aureus, Bacillus Proteus is obviously, the minimum inhibitory concentrations were 0.125 g/mL, 0.125 g/mL, 1 g/mL respectively and no inhibitory effect on Serratieae. [Conclusions] The anthraquinones from Cassia seed can inhibit a part of spoilage bacteria in spiced pig heads.

Antibacterial Activity of Anthraquinone from Aloe on Spiced Pig Head

[Objective] To optimize the extraction of anthraquinone from Aloe by ultrasonic extraction and its antibacterialactivity. [Method]The influences of different extraction time and ethanol concentration, on anthraquinone contentwere evaluated by asingle factor experiment. And anthraquinone content was determined by ultraviolet spectrophotometry. The bacteriostasis of anthraquinone on spiced pig head’s common putrefying bacteria: Staphylococcus, Serratieae, Bacillus, Proteus and the minimal inhibitory concentration (MIC) were studied by oxford plate assay system. [Result]The best extraction time was 30 minutes and the best ethanol concentration was 80%. The antibacterial activity of the Aloe anthraquinone on Staphylococcus Aureus, Bacillus Proteus is obviously, the minimum inhibitory concentrations were 0.0625 g/mL, 0.05 g/mL, 0.125 g/mL respectively and no inhibitory effect on Serratieae. [Conclusions] The anthraquinones from Aloe can inhibit a part Of spoilage bacteria inspiced pig heads.

Water availability effects on plant growth, seed yield, seed quality in Cassia obtusifolia L., a medicinal plant

The seeds of Cassia obtusifolia L. are widely used in traditional Chinese medicine and health tea. The objective of this study was to investigate the effect of water availability on plant growth, seed yield, and seed quality in C. obtusifolia for improving its cultivation. Pot and field irrigation experiments were conducted in Qingdao of China in 2013 and 2014, respectively. Seven water irrigation treatments including 100%, 90%, 80%, 70%, 60%, 50% and 40% of field capacity (FC) in pot experiments while five water irrigation treatments including 100%, 85%, 70%, 55% and 40% of field capacity (FC) in field experiments were applied on the plant of C. obtusifolia, respectively. In both experiments, the results showed that different drought stress significantly resulted in different reduction in plant growth and seed yield attributes of C. obtusifolia, and more pronounced with the severity of drought. However, as for the seed quality attributes, both results showed that weak and moderate drought stress significantly increased the content of anthraquinones (plant secondary metabolites) while significantly decreased the content of protein (plant primary metabolites) in the seeds of C. obtusifolia, except that strong drought stress significantly decreased both of them. Notably, the weak drought stress (70% FC in pot and in field) did not decrease the seed yield significantly which resulted in a biggest harvest index among other treatments and thus resulted in the highest anthraquinones yields per plant. In addition, results showed that strong and moderate drought stress affected the proportion between hydrophilic and lipophilic anthraquinones in seed of C. obtusifolia significantly. Our study suggested that a certain of deficit irrigation has the potential to compromise seed yield and seed quality of C. obtusifolia, which can be used as a practical irrigation strategy for cultivation improving of C. obtusifolia as a medicinal plant.

Simultaneous Determination of 10 Anthraquinones in Rhubarb Based on HPLC-Q-HR/MS

ObjectiveTo develop a simple, sensitive, and precise method for simultaneous determination of 10 anthraquinones in Rhubarb. MethodsHPLC-Q-HR/MS was employed for simultaneous quantification of free anthraquinones (aloe-emodin, emodin, chrysophanol, physcion, and rhein) and their glycosides. Chromatographic analysis was performed on an XBridge™ C18 column (2.1 mm × 150 mm, 5 µm) with mobile phases consisting of 3 mmol/L ammonium acetate (A) and methanol (B) at a flow rate of 0.3 mL/min. ResultsAll calibration curves exhibited good linear relationship (R2 > 0.999). The limits of detection (LOD) and quantification (LOQ) were in the range of 0.39-2.97 ng/mL and 0.56-8.90 ng/mL, respectively. The overall intra- and inter-day precisions of analytes presented as relative standard deviations (RSDs) were less than 2.79%. Relative recoveries varied between 97.83% and 104.28%. The validated method was applied to assess the quality of Rhubarb collected from different regions of China. Results showed that chrysophanol and rhein-8-O-β-D-glucoside was the largest portion of free anthraquinones and anthraquinone glycosides in Rhubarb, respectively. The total content of anthraquinones was higher in Rhubarbs from Sichuan, Qinghai, Yunnan, and Gansu provinces than that in those from Shandong and Henan provinces, while no significant variability existed in different regions of the same province. ConclusionHPLC-Q-HR/MS method is accurate and reliable for simultaneous quantification of above free anthraquinones and their glycosides in Rhubarb and can be applied to standardize the quality of Rhubarb and its quality control in different regions.

The Extracts of Morinda officinalis and Its Hairy Roots Attenuate Dextran Sodium Sulfate-Induced Chronic Ulcerative Colitis in Mice by Regulating Inflammation and Lymphocyte Apoptosis.

Morinda officinalis is beneficial for the treatment of inflammatory bowel disease (IBD). The hairy root with higher genetic and biochemical stability cultured from M. officinalis might have similar effects to treat IBD. In this study, the main chemical composition of the root extracts of M. officinalis (MORE) native plant and the hairy root extract of M. officinalis (MOHRE) was compared by quantitative HPLC. The difference of their therapeutic effects and potential mechanism was evaluated using 3% dextran sodium sulfate-induced chronic colitis in mice and T lymphocytes in vitro. The results found that MOHRE possesses many specific peaks unobserved in the chromatogram of native plant. The content of iridoids in the MORE (3.10%) and MOHRE (3.01%) is somewhat similar but quite different for their anthraquinones’s content (0.14 and 0.66%, respectively). Despite all this, treatment with both MORE and MOHRE significantly attenuated the symptoms of colitis, including diarrhea, body weight loss, colon shortening, histological damage, and decreased inflammatory cytokine levels. In addition, they dose-dependently increased the apoptosis of T lymphocyte in vivo and in vitro. And, the differences for treatment effects on ulcerative colitis (UC) between them both in this study were mostly insignificant. The results demonstrated that the effects of MORE and MOHRE for the treatment of UC are similar, although there are a few difference on their chemical composition, indicating the hairy root cultured from M. officinalis might be able to replace its native plant on treatment of UC. The successful derivation of a sustainable hairy root culture provides a model system to study the synthetic pathways for bioactive metabolites, which will make the use of bioreactors to largely produce traditional medicine become reality.

Preventive effects of Polygonum multiflorum on glucocorticoid-induced osteoporosis in rats.

In Traditional Chinese Medicine, Polygonum multiflorum (PM) is known for its anti-aging properties. A previous study by our group showed that extracts of PM were able to prevent and treat bone loss in vivo, and the active components emodin and 2,3,5,4,-tetrahydroxystilbene-2-O-β-glucoside (TSG) promoted the osteogenic differentiation of mesenchymal stem cells in vitro. The aim of the present study was to investigate the preventive effects of PM on glucocorticoid-induced osteoporosis (GIO) in rats. A crude extract of PM was prepared with 75% ethanol, purified and enriched using a D-101 macroresin column and elution with 30% ethanol, and the material obtained was assessed by high-performance liquid chromatography. Male or female Sprague Dawley rats (n=180) were randomly divided into nine groups: Control, prednisone, prednisone plus calcitriol (CAL), prednisone plus 30% ethanolic eluate of PM [high (H), medium (M) and low (L) dose] and prednisone plus crude extract of PM (H, M and L dose). Prednisone was orally administered to the osteoporosis model rats for 21 weeks, alongside which they received PM extracts. The weight of the viscera, anterior tibial muscle and other tissues was recorded at the end of the experiment. The femur and lumbar vertebra were collected for the measurement of three-dimensional microarchitecture by micro-computed tomography scanning, assessment of biomechanical properties and determination of bone mineral density (BMD). In the 30% ethanolic eluate of the PM extract, the content of TSG and combined anthraquinone was 9.20 and 0.15%, respectively, and that in the crude extract of PM was 2.23 and 0.03%, respectively. Over 6 weeks, the weight of the rats the in prednisone group decreased (P<0.05), while the weight of rats treated with M and H doses of 30% ethanolic eluate was increased compared with that in the prednisone group (P<0.05). Rats exposed to prednisone exhibited a deteriorated bone microarchitecture, low BMD, decreased bone volume/total volume and poor biomechanical properties. Furthermore, the weight of the adrenal gland and the anterior tibial muscle was decreased. 30% ethanolic eluate of PM at M and L doses and crude extract of PM at the H dose counteracted the alterations of skeletal and other characteristics induced by prednisone in rats, as did CAL. In conclusion, extracts of PM exerted a protective effect on bone tissue in GIO rats.

Quality evaluation of rhubarb dispensing granules based on multi-component simultaneous quantitative analysis and bioassay

This study attempts to evaluate the quality of Chinese formula granules by combined use of multi-component simultaneous quantitative analysis and bioassay. The rhubarb dispensing granules were used as the model drug for demonstrative study. The ultra-high performance liquid chromatography (UPLC) method was adopted for simultaneously quantitative determination of the 10 anthraquinone derivatives (such as aloe emodin-8-O-β-D-glucoside) in rhubarb dispensing granules; purgative biopotency of different batches of rhubarb dispensing granules was determined based on compound diphenoxylate tablets-induced mouse constipation model; blood activating biopotency of different batches of rhubarb dispensing granules was determined based on in vitro rat antiplatelet aggregation model; SPSS 22.0 statistical software was used for correlation analysis between 10 anthraquinone derivatives and purgative biopotency, blood activating biopotency. The results of multi-components simultaneous quantitative analysisshowed that there was a great difference in chemical characterizationand certain differences inpurgative biopotency and blood activating biopotency among 10 batches of rhubarb dispensing granules. The correlation analysis showed that the intensity of purgative biopotency was significantly correlated with the content of conjugated anthraquinone glycosides (P<0.01), and the intensity of blood activating biopotency was significantly correlated with the content of free anthraquinone (P<0.01). In summary, the combined use of multi-component simultaneous quantitative analysis and bioassay can achieve objective quantification and more comprehensive reflection on overall quality difference among different batches of rhubarb dispensing granules.

Impact of phosphorus and potassium fertilizers on growth and anthraquinone content in Rheum tanguticum Maxim. ex Balf

Abstract The dried root of Rheum tanguticum plays an important role in formulations and prescriptions in traditional Chinese medicine and Kampo medicine. Due to over-exploitation, R. tanguticum resources have decreased sharply in recent years. The main objective of our investigation (a 3-year field experiment) was to explore the effect of different levels of phosphorus (superphosphate) and potassium (potassium sulfate) fertilizer on the biomass (root fresh weight, root increment, and root dry weight), yield, dry matter content, and anthraquinone content of this plant at different harvesting stages (green stage, growth stage, and wilting stage) under alpine conditions. The root fresh weight and root dry weight increased significantly at the wilting stage following treatment with 90 kg P2O5/ha (100% and 59%, respectively) in 2016 and 75 kg K2O/ha (43% and 41%, respectively) in 2015 compared to the control. The yield of root dry weight obtained from three-year-old R. tanguticum plants was 9200 kg/ha when 90 kg P2O5/ha of phosphorus fertilizer was applied, and 10,400 kg/ha when 75 kg K2O/ha of potassium fertilizer was applied. This yield reached a maximum at the wilting stage. The anthraquinone content of two-year-old R. tanguticum plants had already reached the standard level of the Chinese Pharmacopoeia; however, three-year-old plants had double the anthraquinone content of two-year-old plants. Phosphorus and potassium fertilizers had no obvious influence on the anthraquinone content of R. tanguticum at the same harvesting stage.

Antioxidant, phenolic and antifungal profiles of Acanthus mollis (Acanthaceae).

Acanthus mollis is used as ornamental and medicinal plant. The ethnopharmacology reports indicate that extracts have anti-inflammatory activity. Phytoconstituents profile was evaluated by estimating the content of anthraquinones, flavonoids and phenols. In addition, the antioxidant activity was evaluated using four methods: Hydrogen atoms transfer (TRAP, ORAC and DPPH assays), and single electron transfer (FRAP assay). Finally, antifungal activity was determined by the M27-A2 test. The results shown that ethanol extracts have the highest concentration of phenols, anthraquinones and flavonoids. Total antioxidant capacity, extracts of ethyl acetate and ethanol are those with the highest activity, which correlates strongly with the presence of phenols. The antifungal activity measured in various strains of Candida is concentrated in ethyl acetate extracts of flower and leaf ethanol, a phenomenon may be related to antioxidant activity.

Safety of cassia gum as a feed additive for dogs and cats based on a dossier submitted by Intercolloid (UK) Ltd.

The additive cassia gum consists mainly of high‐molecular weight polysaccharides composed primarily of a linear chain of 1,4‐β‐d‐mannopyranose units with 1,6‐linked α‐d‐galactopyranose units. In 2014, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) delivered an opinion on the safety and efficacy of cassia gum. The FEEDAP Panel concluded, based on positive findings observed in a bacterial reverse mutation test with a semirefined cassia gum (about 70 mg anthraquinones/kg) but not with purified semirefined cassia gum that meets the specification as a food additive (< 0.5 mg anthraquinones/kg), that only purified semirefined cassia gum that meets the specifications of cassia gum as a food additive can be considered safe for cats and dogs, at a maximum content of 1.5% cassia gum (15,000 mg/kg feed) in dry matter, corresponding to 1.32% (13,200 mg/kg feed) in a standardised complete feed with 12% water content. Following this opinion, the European Commission gave the possibility to the applicant to submit complementary information in order to complete the assessment on the safety for all animal species. The new data submitted, describing a reproduction study in cats, do not address the potential of the additive to exert mutagenic effects, and do not allow conclusions to be drawn on the safety of semirefined cassia gum with an anthraquinone content higher than the one specified for the use as a food additive (< 0.5 mg/kg). The FEEDAP Panel is not in the position to modify the conclusions reached in the previous opinion.

Callus culture and in vitro production of anthraquinone in Gynochthodes umbellata (L.) Razafim. & B. Bremer (Rubiaceae)

Gynochthodes umbellata is a rare medicinal plant used in traditional systems of medicine for curing various ailments. It is a woody climber belonging to the family Rubiaceae. Its over collection from the wild population resulted the rapid depletion of this species. There is an urgent need to conserve this species and also establish an alternative system for the production of bioactive compound in vitro without harming the wild population. The present investigation was carried out to analyse secondary metabolite (anthraquinone) production in callus cultures of G. umbellata. In vitro production of anthraquinone through callus cultures of G. umbellata was standardized using in vitro leaves, derived from the nodal explant cultures maintained in Murashige and Skoog solid medium containing 2mg/l 6-benzylaminopurine and 3% sucrose. Leaves were harvested from the healthy shoots of axenic cultures and culture conditions were optimised for the establishment of callus. In vitro derived leaves (1–2cm2) were inoculated onto Murashige and Skoog medium containing different individual concentrations of 2, 4-Dichlorophenoxyacetic acid (0.5–5mg/l), indole-3-butyric acid (0.5–5mg/l), indole-3-acetic acid (0.5–5mg/l) and 1-naphthaleneacetic acid (0.5–5mg/l). Of the different auxins tried for callus induction, 2, 4-Dichlorophenoxyacetic acid was found to be the most suitable auxin with different concentrations for the induction of friable yellow callus. 1-Naphthaleneacetic acid doesn’t show any callus formation while indole-3-butyric acid and indole-3-acetic acid produced greenish white compact callus. A maximum fresh weight (1.7953±0.61g) and dry weight (0.1149±0.04g) of the friable yellow callus and highest amount of anthraquinone content (18.1875±0.58mg/g dw) were recorded in MS medium supplemented with 1mg/l 2, 4-Dichlorophenoxyacetic acid. From the present study it revealed that the callus culture is an effective method for the production of anthraquinone in vitro in G. umbellata. Anthraquinone production through in vitro leaf segment derived callus culture is a viable system for the production of this pharmacologically and industrially important natural plant pigment.

Quality evaluation of Heshouwu, a Taoist medicine in Wudang, China.

Polygonum multiflorum Thunb., which is known as Heshouwu in China, is a Taoist medicine sourced from the Wudang mountain area. At present, the quality of the Heshouwu sourced from this region is unknown. The present study aimed to evaluate the quality of wild Heshouwu collected from the Wudang mountain area, particularly the 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) and combined anthraquinone (CAQ) content, compared with that of commercially available Heshouwu. Furthermore, the potential quantities of organic pesticide residues were determined. High performance liquid chromatography with a diode array detector was used to quantify TSG and CAQ content, whereas gas chromatography (GC), performed using a temperature gradient, was used to detect the presence of organochlorine, pyrethroid and organophosphorus pesticides. The average TSG content present in the wild Heshouwu from the Wudang mountain area and in the commercially available Heshouwu was 2.39 and 1.10%, respectively. In addition, the average content of CAQ in these was 1.41 and 3.46%, respectively. GC did not detect residues of organic pesticides in the wild Heshouwu, thus this plant met the criterion of the Chinese Pharmacopeia (2010 edition). The results of the present study indicated that wild Heshouwu from the Wudang mountain area may be suitable for use as a Chinese medicine across China.

Anthraquinones isolated from the browned Chinese chestnut kernels (Castanea mollissima blume)

Anthraquinones (AQS) represent a group of secondary metallic products in plants. AQS are often naturally occurring in plants and microorganisms. In a previous study, we found that AQS were produced by enzymatic browning reaction in Chinese chestnut kernels. To find out whether non-enzymatic browning reaction in the kernels could produce AQS too, AQS were extracted from three groups of chestnut kernels: fresh kernels, non-enzymatic browned kernels, and browned kernels, and the contents of AQS were determined. High performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) methods were used to identify two compounds of AQS, rehein(1) and emodin(2). AQS were barely exists in the fresh kernels, while both browned kernel groups sample contained a high amount of AQS. Thus, we comfirmed that AQS could be produced during both enzymatic and non-enzymatic browning process. Rhein and emodin were the main components of AQS in the browned kernels.

Induction of hairy roots and over production of anthraquinones in Oldenlandia umbellata L.: a dye yielding medicinal plant by using wild type Agrobacterium rhizogenes strain

Hairy root cultures of Oldenlandia umbellata L. was established by infecting leaf explants with wild type Agrobacterium rhizogenes strain (MTCC 532). Leaf explants were co-cultured with activated bacteria in half strength Murashige and Skoog (MS) basal medium containing 500 mg l−1 ampicillin for the induction of hairy roots. To facilitate effective transfer of T-DNA of Ri plasmid, duration of co-cultivation and concentration of acetosyringone were standardized as 48 h and 200 µM, respectively. Roots were produced from the infected leaves within 14 days. Fast growing roots (2–3 cm) were excised and sub-cultured in half strength basal MS liquid medium for the development of transgenic lines. Thus ten hairy root lines were raised and superior line L3 was selected on the basis of fresh weight (1.5 g), dry weight (0.26 g) and anthraquinone (AQ) content (10.374 mg g−1 dry wt). Superior line (L3) was further cultured to test stability of AQ production and found that stable production of AQ during five consecutive sub-cultures. High performance liquid chromatographic method was adopted for quantification of purpurin from samples. Hairy roots produced 21.83 mg g−1 purpurin, while roots from wild growing plants produced 6.08 mg g−1 purpurin. Thus purpurin content in hairy roots was about 3.6 times higher than wild growing roots. Further, polymerase chain reaction based amplification using primers specific for genes rolA, rolB, and rolC confirmed the transformation.

Reduction of Candida tropicalis biofilm by photoactivation of a Heterophyllaea pustulata extract

Context: Biofilm formation is an important problem, since this growth mode confers resistance to drugs usually used in therapeutics.Objective: In vitro antifungal activity of extracts obtained from Heterophyllaea pustulata Hook f. (Rubiaceae) were studied against Candida tropicalis biofilms, evaluating the effect of irradiation and the oxidative and nitrosative stresses as possible mechanisms of action.Materials and methods: Hexane, benzene, ethyl acetate and ethanol extracts were evaluated at three concentrations (0.2, 0.1 and 0.05 mg/mL) over mature biofilm, under darkness and irradiation. After 48 h of incubation, biofilm quantitation was performed by the O'Toole and Kolter method. Reactive oxygen species (ROS) was measured by nitro-blue tetrazolium (NBT) reaction and reactive nitrogen intermediates (RNI) by the Griess reagent. Superoxide dismutase activation (SOD, NBT assay) and total antioxidant system (FRAP test) were studied.Results: Only the benzene extract at 0.2 mg/mL reduced the biofilms formation. The slight decrease achieved in darkness (17.06 ± 2.80% reduction) was increased by light action (39.31 ± 3.50% reduction), clearly observing a photostimulation. This great reduction was confirmed by confocal microscopy. In darkness, biofilm reduction was mediated by an increase in RNI, whereas under irradiation, the ROS action was most important. Although no SOD activation was observed, a strong stimulation of the total antioxidant system was detected. HPLC analysis established a high content of several anthraquinones in this extract.Discussion and conclusion: Biofilm reduction by benzene extract was mainly mediated by oxidative stress triggered under light action, confirming a photodynamic sensitization, which could be attributed to its high content of photosensitizing anthraquinones.

Increase of anthraquinone content in Rubia cordifolia cells transformed by native and constitutively active forms of the AtCPK1 gene.

Overexpression of both native and mutant forms of AtCPK1 in Rubia cordifolia cells increased anthraquinone production and transcript abundance of the RcIPPI, RcOSBL, RcOSBS , and RcICS genes to different extents. Calcium-dependent protein kinases (CDPKs) are involved in various cell processes and are regulated by a calcium signal system. CDPKs also function in plant defense against stress factors such as pathogens, temperature, and salinity. In this study, we compared the effect of heterologous expression of two forms of the Arabidopsis AtCPK1 gene, native and constitutively active (Ca(2+)-independent), on anthraquinone production in transgenic Rubia cordifolia cells. Significant qualitative and quantitative differences were found in the content of anthraquinone derivatives in control and AtCPK1-transgenic calli. Expression of the AtCPK1 gene increased anthraquinone production by 3 and 12 times for native and constitutively active forms, respectively, compared with control cells. In addition, we identified and quantified the expression of genes encoding key enzymes of the anthraquinone biosynthesis pathway, including isochorismate synthase (ICS), o-succinylbenzoate synthase (OSBS), o-succinylbenzoate ligase (OSBL), and isopentenyl diphosphate isomerase (IPPi). In all AtCPK1-transgenic cell lines, expression of ICS, OSBS, OSBL, and IPPi increased considerably at 14-15days of subculture and decreased at the end of cultivation (30days). The results suggest that both native and constitutively active AtCPK1 forms induced anthraquinone accumulation at the logarithmic growth stage via enhancement of expression of genes involved in the metabolism of anthraquinones or their regulatory mechanisms.

Comparative Histological and Phytochemical Study of Fallopia Species

Fallopia species which belong to the Polygonaceae family have several data related to their use in the Asian herbal medicine. In this work, some histological and phytochemical parameters of Fallopia japonica, F. sachalinensis, and F. x bohemica were analysed and compared. Rhizome and leaf samples were collected before, during, and after the flowering period at 3 habitats in Szombathely and 4 habitats in Baranya County, Hungary. The main histological characteristics of the stem, leaf and petiole were studied by light microscopy in cross section. Total tannin and anthraquinone contents were determined according to the official methods of the Hungarian Pharmacopoeia VIIIth (equal to the European Pharmacopoeia 6th). No species-specific markers were found in any plant part. In the rhizome, the highest tannin content was measured in Japanese knotweed, followed by Bohemian and giant knotweed in each period. The tannin content measured in each plant was higher in the leaves than in the rhizomes except F. japonica. The rhizome of F. japonica had the highest anthraquinone content before the flowering period, followed by F. x bohemica and F. sachalinensis. According to earlier and our preliminary data, Fallopia taxa are of great therapeutic promise in the future.

Synergistic effect of methyl jasmonate and cyclodextrins on anthraquinone accumulation in cell suspension cultures of Morinda citrifolia and Rubia tinctorum

Plant in vitro culture is a platform for producing secondary metabolites that combines safety, quality and low environmental impact. Besides, it is possible to increase the accumulation of these compounds by different strategies, such as elicitation. In this work, we analyzed the effects of the combination of methyl jasmonate (MeJ) and two cyclodextrins (CDs) on the production of anthraquinones (AQs) in cell cultures of Rubiaceae (Morinda citrifolia and Rubia tinctorum). These secondary metabolites have been traditionally used as dyes and have interesting therapeutic applications. The experiments were designed according to a full factorial design of two factors (MeJ and a CD) in two levels (0 and 0.1 mM for MeJ, and 0 and 20 mM of the CD). MeJ and (2-hydroxypropyl)-β-cyclodextrin (HPCD) synergistically increased intracellular AQ content in suspension cultures of R. tinctorum, and, to a lesser extent, in suspension cultures of M. citrifolia. Combination of MeJ with another CD, 2-methyl-β-cyclodextrin, led to a more intense and later increase in AQ content in cell cultures of R. tinctorum when compared to MeJ–HPCD treatment. However, the combination of CD and MeJ failed to induce a drastic AQ release to the culture media. Nevertheless, our results show that combination of strategies (using a CD and MeJ) was successful to increase secondary metabolite accumulation in suspension cultures. To our knowledge, this is the first report of synergistic effect of MeJ and CD on AQ accumulation in plant in vitro cultures.

Research on contents of anthraquinones in Cassiae Semen by principal component analysis

Cassiae Semen is a common traditional Chinese medicine, and contents of anthraquinones of Cassiae Semen different significantly from area to area. According to Chinese Pharmacopoeia (2010 edition), only contents of aurantio obtusin and chrysophanol were used to evaluate the quality of Cassiae Semen, another data could be added later. Ten batches of Cassiae Semen from different areas were determined, and total anthraquinones, total free anthraquinones and total combined anthraquinones contents were assessed by ultraviolet visible spectrophotometer, contents of aurantio obtusin, rhein, aloe emodin, emodin, chrysophanol and physcion were determined by HPLC. After that, principal components analysis was used to evaluate these data determined previous by dimension reduction analysis. At last, the result suggests that three main components were found out, it shows that content of aloe emodin could be used to evaluate the quality of Cassiae Semen as well as contents of aurantio obtusin and chrysophanol. And Cassiae Semen from Hebei province posseses higher quality than Cassiae Semen from other different areas. All these results can provide a good reference for quality evaluating of Cassiae Semen medicinal materials at a certain extent.