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Chrysanthemum indicum Linné has been used in traditional medicine to treat various inflammatory diseases in East Asia. The aim of the present study was to investigate the protective effect of C. indicum ethanol extract (CILE) against cisplatin-induced nephrotoxicity. An HPLC-photodiode array method was used for fingerprint analysis of the CILE and ten major constituents were quantitatively analyzed. The protective effect of CILE on cisplatin-induced nephrotoxicity was assessed using both in vitro (porcine kidney cell; PK15 cell) and in vivo (Sprague Dawley rat) experiments. In the in vitro study, CILE enhanced PK15 cell viability after cisplatin treatment with recovered antioxidant status. Moreover, the increased p53 expression after cisplatin treatment was decreased in the CILE pretreated cells. In the in vivo study, SD rats were treated for 28 consecutive days with CILE (0, 100, 300 and 500 mg/kg). On day 23, a single dose of cisplatin (5 mg/kg) was injected to induce nephrotoxicity. The CILE pretreated group showed recovered serum renal function index with ameliorated oxidative stress. Histopathological alterations and apoptosis in the kidney were also decreased in CILE pretreated rats. Taken together, CILE could attenuate cisplatin-induced nephrotoxicity and might be a beneficial agent for acute renal failure management.

Roxarsone (ROX), a world widely used feed organoarsenic additive in animal production, can be excreted as itself and its metabolites in animal manure. Animal manure is commonly land applied with phosphorous (P) fertilizer to enhance the P phytoavailability in agriculture. We investigated the accumulation of As species in garland chrysanthemum (C. coronarium) plants fertilized with 1% (w/w, manure/soil) chicken manure bearing ROX and its metabolites, plus 0, 0.05, 0.1, 0.2, 0.4, and 0.8 g P2O5/kg, respectively. The results show that As(III) was the sole As compound in garland chrysanthemum shoots, and As(III) and As(V) were detectable in roots. Elevated phosphate level supplied more As(V) for garland chrysanthemum roots through competitive desorption in rhizosphere, leading to significantly enhanced accumulation of As species in plants. As(III) was the predominant As form in plants (85.0∼90.6%). Phosphate could not change the allocation of As species in plants. Hence, the traditional practice that animal manure is applied with P fertilizer may inadvertently increase the potential risk of As contamination in crop via the way ROX → animal → animal manure → soil → crop.

Objective: To provide a scientific basis for quality control of "Boju" Chrysanthemum morifolium by establishing a HPLC characteristic fingerprint., Methods: The HPLC analysis was performed on a Spursil C18 chromatographic column (250 mm x 4. 6 mm, 5 µm), and the mobile phase was acetonitrile-0. 1% phosphoric acid in a gradient mode with the flow rate of 1. 0 mL/min. The detection wavelength was 325 nm and the temperature of column was 30 °C., Results: The common pattern of HPLC characteristic chromatographic profile was established. There were 16 common peaks, five of which were identified in the pattern. The similarities of 10 batches of "Boju" Chrysanthemum monrfolium were evaluated, and all of them were greater than 0. 900., Conclusion: This analysis method of HPLC characteristic chromatographic fingerprint is simple and reproducible, and it can provide a scientific basis for identification and quality evaluation of "Boju" Chrysanthemum monfolium.

Chrysanthemum is an important traditional Chinese medicine and is drunk daily as a herbal tea. Chlorogenic acids and flavonoids are generally considered as the bioactive compounds. In this work, six kinds of Juhua Tea were analyzed and their active compounds and antioxidant activities were compared. In total, 32 phenolic compounds were profiled and identified using HPLC-DAD/ESI-MSn, composed of chlorogenic acids (10), flavones (8), chalcones (8), flavanones (4) and flavonols (2). Chalcones were the main flavonoids in Kunlun Xueju (Coreopsis tinctoria) extract, while flavones and chlorogenic acids were dominant in the five Chrysanthemum teas. Total chlorogenic acids content (TCA) was highest in Tai Ju (Chrysanthemum morifolium cv. 'Tai Ju') (8.59 ± 0.87 mg/g DW), and total flavonoids content (TF) was highest in Kunlun Xueju (87.2 ± 7.0 mg/g DW), which were both lowest in Ganye Ju (Chrysanthemum eticuspe) (TCA 0.86 ± 0.26 mg/g DW, TF 1.43 ± 0.41 mg/g DW). Huangin Ju (Anthemis tinctoria) possessed the most flavones (19.7 ± 0.6 mg/g DW). Antioxidant capacity of each drink, assessed by Folin-Ciocalteu, DPPH, ABTS and FRAP assays, consistently showed that Kunlun Xueju extract possessed stronger antioxidant activity than the other five, suggesting that the flavonoids content accounted for the free radical scavenging. The present work provides a method for the characterization and quality control of Juhua Tea. Moreover, it is a guideline for consuming choice, due to the different biological functions resulting from chalcones, chlorogenic acids, and flavones.

Objective: To study the chemical constituents from the leaves of "Chuju" Chrysanthemum morifolium., Methods: All compounds were separated and purified by column chromatography over silica gel, Sephadex LH-20 and preparative HPLC. Their structures were identified by spectral methods including 1H-NMR and 13C-NMR., Results: 21 compounds were isolated and identified as octa-cosyl alcohol (1), β-sitosterol (2), lupeol (3), α-amyrin (4), daucosterol (5), ineupatorolide B (6), syringin (7), chlorogenic acid (8), petasiphenol (9), physcion (10), acacetin (11), eupatilin (12), quercetin (13), diosmetin (14), luteolin (15), apigenin (16), apigenin- 7-O-β-D-glucopyranoside (17), quercetin-3-O-β-D-glucopyranoside (18), luteolin-7-O-β-D-gluco pyranoside (19), apigenin-7-O-β-D- neospheroside (20), and acacetin-7-O-β-D-glucoside (21)., Conclusion: Compounds 1-12, 18 and 20 are isolated from this plant for the first time. Compounds 10, 13, 14, 15 and 16 have shown strong antioxidant activities by DPPH · scavenging activity better than Vit C.

A simple sample pretreatment for simultaneous determination of nine fungicides (triadimefon, picoxystrobin, kresoxim-methyl, diniconazole, epoxiconazole, trifloxystrobin, triticonazole, difenoconazole, and azoxystrobin) in chrysanthemum was developed using matrix extraction-vortex-assisted dispersive liquid-liquid microextraction (ME-VADLLME) prior to gas chromatography with electron capture detection. The target fungicides were firstly extracted with acetonitrile and cleaned with the mixture of primary secondary amine and graphite carbon black. The VADLLME procedure was performed by using toluene with lower density than water as the extraction solvent and the acetonitrile extract as the dispersive solvent, respectively. After vortexing and centrifugation, the fine droplet of toluene was collected on the upper of the mixed toluene/acetonitrile/water system using a 0.1-mL pipettor. Under the optimum conditions, the relative recoveries ranged from 73.9 to 95.1% with relative standard deviations of 3.5-9.7% for all of the analytes. The limits of detection were in the range of (0.005-0.05)×10(-3) mg kg(-1). In the proposed method, the ME step provides more effective cleanup for the chrysanthemum matrix, and VADLLME introduces higher sensitivity with the remarkable enrichment factors up to 88-fold compared with the conventional QuEChERS or SPE. The good performance has demonstrated that ME-VADLLME has a strong potential for application in the multi-residue analysis of complex matrices., (Copyright © 2014 Elsevier B.V. All rights reserved.)

One new octulosonic acid derivative, chrysannol A (1), along with 17 known compounds (2-18), were isolated from Chrysanthemum indicum flowers. Their structures were determined from 1D NMR, 2D NMR, HR-ESI-MS spectral data, and comparisons with previous reports. The effects of these compounds on lipopolysaccharide (LPS)-induced nitric oxide (NO) and tumor necrosis factor alpha (TNF-α) production by RAW 264.7 cells were investigated. Compound 8 showed the highest inhibition of NO production of 46.09% at a concentration of 10.0μM. Compounds 7, 10, 11, and 16 inhibited TNF-α secretion at all concentration tested (0.4, 2.0, and 10.0μM), with inhibition values ranging from 22.27% to 33.13%. In addition, compound 8 and 9 decrease COX-2 and iNOS protein on Western blot analysis in dose dependent manner., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

For the determination of seven caffeoylquinic acids [neochlorogenic acid (NcA), cryptochlorogenic acid (CcA), chlorogenic acid (CA), caffeic acid (CfA), isochlorogenic acid A (Ic A), isochlorogenic acid B (Ic B), isochlorogenic acid C (Ic C)] and two flavonoids [luteolin 7-O-glucoside (LtG) and luteolin (Lt)], a three-channel liquid chromatography with electrochemical detection (LC-3ECD) method was established. Chromatographic peak heights were proportional to each concentration, ranging from 2.5 to 100 ng/mL for NcA, CA, CcA, and CfA, and ranging from 2.5 to 250 ng/mL for LtG, Ic B, Ic A, Ic C, and Lt, respectively. The present LC-3ECD method was applied to the quantitative analysis of caffeoylquinic acids and flavonoids in four cultivars of Chrysanthemum morifolium flowers and their sulfur-fumigated products. It was found that 60% of LtG and more than 47% of caffeoylquinic acids were lost during the sulfur fumigation processing. Sulfur fumigation showed a destructive effect on the C. morifolium flowers. In addition, principle component analyses (PCA) were performed using the results of the quantitative analysis of caffeoylquinic acids and flavonoids to compare the "sameness" and "differences" of these analytes in C. morifolium flowers and the sulfur-fumigated products. PCA score plots showed that the four cultivars of C. morifolium flowers were clearly classified into four groups, and that significant differences were also found between the non-fumigated C. morifolium flowers and the sulfur-fumigated products. Therefore, it was demonstrated that the present LC-3ECD method coupled with PCA is applicable to the variation analysis of different C. morifolium flower samples.

In this study, the antidiabetic effects of a hot water extract of edible Chrysanthemum morifolium Ramat. (HW-ECM) were investigated in type 2 diabetic mice. HW-ECM improved blood glucose levels and insulin resistance and increased adiponectin mRNA expression in adipose tissues and protein concentrations in the plasma. Moreover, it increased adipose mRNA and protein expressions of peroxisome proliferator-activated receptor γ (PPARγ), a regulator of adiponectin transcription, and mRNA expression of its downstream target genes. It also reduced the adipose cell size and attenuated the mRNA expression of pro-inflammatory adipocytokines in adipose tissues. These data presumably indicate a hypoglycemic mechanism of HW-ECM, involving increased PPARγ expression, decreased the adipocyte sizes, and suppression of chronic inflammation in adipose tissues. Finally, elevated adiponectin levels lead to amelioration of insulin resistance and the corresponding hypoglycemic effects. Therefore, HW-ECM indicates its potential as a functional food for type 2 diabetes.

The adipose tissue is an endocrine organ, and its endocrine function is closely related to type 2 diabetes. Edible Chrysanthemum morifolium Ramat. (ECM) possesses several biological properties; however, its effect on adipocytes remains unclear. We investigated the effect of the hot water extract of ECM (HW-ECM) on 3T3-L1 adipocytes. HW-ECM enhanced adipocyte differentiation, adiponectin secretion, and glucose uptake in 3T3-L1 cells. It also increased the mRNA levels of peroxisome proliferator-activated receptor γ (PPARγ), a regulator of adipocyte differentiation, adiponectin transcription, and GLUT4 expression. In addition, HW-ECM increased the mRNA levels of CCAAT/enhancer-binding protein-delta (C/EBPδ), which induces PPARγ expression, but not C/EBPβ, during early adipocyte differentiation. These results suggest that HW-ECM enhances adipocyte differentiation, adiponectin secretion, and glucose uptake through C/EBPδ-induced PPARγ expression. These effects of HW-ECM on adipocytes suggest that HW-ECM is potentially beneficial for type 2 diabetes.

We have proposed an assessment methods to estimate the measurement relative standard deviation (RSD) of chromatographic peaks in quantitative HPLC for herbal medicines by the methodology of ISO 11843 Part 7 (ISO 11843-7:2012), which provides detection limits stochastically. In quantitative HPLC with UV detection (HPLC-UV) of Scutellaria Radix for the determination of baicalin, the measurement RSD of baicalin by ISO 11843-7:2012 stochastically was within a 95% confidence interval of the statistically obtained RSD by repetitive measurements (n = 6). Thus, our findings show that it is applicable for estimating of the repeatability of HPLC-UV for determining baicalin without repeated measurements. In addition, the allowable limit of the "System repeatability" in "Liquid Chromatography" regulated in a pharmacopoeia can be obtained by the present assessment method. Moreover, the present assessment method was also successfully applied to estimate the measurement RSDs of quantitative three-channel liquid chromatography with electrochemical detection (LC-3ECD) of Chrysanthemi Flos for determining caffeoylquinic acids and flavonoids. By the present repeatability assessment method, reliable measurement RSD was obtained stochastically, and the experimental time was remarkably reduced.

The objective of the present study was to evaluate an efficient purifying process of Kunlun Chrysanthemum procyanidins (KCPC) by combination of AB-8 macroporous adsorption resin and silica gel column through adsorption and desorption experiments under static and dynamic status with the purity of procyanidins and the antioxidant activity as indexes respectively. The optimum parameters for adsorption by AB-8 resin were as follows: Sample concentration 1mg/mL, pH6, injecting velocity 2 bed volume (BV) /h, with 3 BV 70% ethanol as eluting solvent, and elution flow rate 2BV /h. One cycle after treatment using AB-8 resin, the purity of KCPC obviously enhanced 3 times compared with that of not prepurified, that is from 22.68 to 63.76%. The optimum parameters for adsorption by silica gel column were as follows: The concentration of procyanidins in a sample solution of 1.2mg/mL (pH 6) with a speed of 2BV/h. Concentration for desorption, with 5 BV of 80% ethanol as an eluent at a flow rate of 2 BV/h. After one-run treatment with silica gel column, the purity of KCPC increased from 63.76 to 81.97%. The antioxidant activities of the purified KCPC in vitro were further investigated. The results indicated that the purification method of combination of AB-8 resin and silica gel column was superior to AB-8 adsorption resin used alone in term of antioxidant activities. Moreover, the 1,1-diphenyl-2-picrylhydrazyl (DPPH•) scavenging ability, the scavenging activity of hydroxyl radicals and the reducing ability appeared to be dose-dependent of KCPC. The novel purification method of combination of AB-8 resin and silica gel column will offer a promising way to purify KCPC for wider application.

Methanol-soluble constituents from the flowers, non-flowering aerial parts and roots of Chrysanthemum pacificum Nakai were analysed via high resolution UPLC-PDA-qTOF-MS followed by chemometrics. Forty-seven chromatographic peaks belonging to various metabolite classes were detected. Most metabolite classes showed qualitative and quantitative differences across parts, with luteolin conjugates being mostly enriched in flowers whereas non-flowering aerial parts contained mostly quercetin and methoxylated flavone conjugates. Root sample ranked the lowest for all flavones and dicaffeoylquinic acids. In contrast, 1,5-di-caffeoylquinic acid levels were found at high levels in flowers and aerial parts reaching 3145 and 1390 μg/g, respectively, suggesting that C. pacificum could serve as a natural resource of this well-recognised anti-hepatotoxic phenolic. Principal component analysis was further used for organs classification in an untargeted manner. This study provides the first map of secondary metabolites distribution in C. pacificum Nakai organs.

The objective of the present study was to evaluate the antioxidant activity, antitumor effect, and antiaging property of proanthocyanidins from Kunlun Chrysanthemum flowers (PKCF) grown in Xinjiang. In vitro antioxidant experiments results showed that the total antioxidant activity and the scavenging capacity of hydroxyl radicals ((•)OH) and 1,1-diphenyl-2-picrylhydrazyl (DPPH(•)) radicals increased in a concentration-dependent manner and were stronger than those of vitamin C. To investigate the antioxidant activity of PKCF in vivo, we used serum, liver, and kidney from mouse for the measurement of superoxide dismutase (SOD), malondialdehyde (MDA), and total antioxidant capacity (T-AOC). Results indicated that PKCF had antioxidative effect in vivo which significantly improved the activity of SOD and T-AOC and decreased MDA content. To investigate the antitumor activity of PKCF, we used H22 cells, HeLa cells, and Eca-109 cells with Vero cells as control. Inhibition ratio and IC50 values were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; PKCF showed great inhibitory activity on H22 cells and HeLa cells. We also used fruit flies as a model for analyzing the anti-aging property of PKCF. Results showed that PKCF has antiaging effect on Drosophila. Results of the present study demonstrated that PKCF could be a promising agent that may find applications in health care, medicine, and cosmetics.

Genetic engineering of roses and other plants of floricultural importance to give them a truly blue petal color is arguably one of the holy grails of plant biotechnology. Toward this goal, bluish carnations and roses were previously engineered by establishing an exclusive accumulation of delphinidin (Dp)-type anthocyanins in their petals via the heterologous expression of a flavonoid 3',5'-hydroxylase gene. Very recently, purple-blue varieties of chrysanthemums were also genetically engineered via a similar biochemical strategy. Although the floral colors of these transgenic plants still lack a true blue color, the basis for the future molecular breeding of truly blue flowers is via the engineering of anthocyanin pathways. Anthocyanins with multiple aromatic acyl groups (often referred to as polyacylated anthocyanins) in the 3'- or 7-position tend to display a more stable blue color than non-acylated anthocyanins. The 7-polyacylation process during the biosynthesis of purple-blue anthocyanins in delphinium (Delphinium grandiflorum) was found to occur in vacuoles using acyl-glucose as both the glucosyl and acyl donor. Glucosyltransferases and acyltransferases involved in anthocyanin 7-polyacylation in delphinium are vacuolar acyl-glucose-dependent enzymes belonging to the glycoside hydrolase family 1 and serine carboxypeptidae-like protein family, respectively. The 7-polyacylation proceeds through the alternate glucosylation and p-hydroxybenzoylation catalyzed by these enzymes. p-Hydroxybenzoyl-glucose serves as the p-hydroxybenzoyl and glucosyl donor to produce anthocyanins modified with a p-hydroxybenzoyl-glucose concatemer at the 7-position. This novel finding has provided a potential breakthrough for the genetic engineering of truly blue flowers, where polyacylated Dp-type anthocyanins are accumulated exclusively in the petals., (© The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

We investigated the effect of essential oil from the flower of Chrysanthemum boreale Makino (CBMEO) on growth of human keratinocytes (HaCaTs) and explored a possible mechanism for this response. CBMEO was extracted using the steam distillation method. CBMEO contained a total of 33 compounds. CBMEO stimulated HaCaT proliferation (EC50, 0.028 μg/mL) and also induced phosphorylation of Akt and ERK1/2 in HaCaTs (EC50, 0.007 and 0.005 μg/mL, for phosphorylated Akt and ERK1/2, respectively). Moreover, CBMEO promoted wound closure in the dorsal side skin of rat tail. This study demonstrated that CBMEO can stimulate growth of human skin keratinocytes, probably through the Akt and ERK1/2 pathways. Therefore, CBMEO may be helpful in skin regeneration and wound healing in human skin, and may also be a possible cosmetic material for skin beauty.

Introduction: The Chrysanthemum genus consisting of about 200 species is mainly distributed over the Northern Hemisphere. Despite the pleasant odour of C. japonense var. debile (setonojigiku), no detailed analysis of the aroma-active compounds has been reported using sensory evaluation., Objectives: Using a hydrodistillation (HD) and a solvent-assisted flavour evaporation (SAFE) method to obtain the volatile oil from the leaf parts., Methods: To clarify odorants contributing to the characteristic aroma-active compounds, the aroma-extract dilution analysis (AEDA) method was performed through gas chromatography olfactometry (GC/O) analysis. In addition, the odour activity value (OAV) was calculated in order to determine the relative contribution of each compound to the aroma-active compounds., Results: A total of 42 components by HD oil were identified by GC-MS, whereas 34 components were identified in SAFE oil. Thirteen compounds were identified by GC/O analysis in HD and SAFE oils respectively., Conclusion: Each extraction method has its own advantages and disadvantages, and they are generally complementary to each other. On the basis of AEDA, OAV and sensory evaluations, [2.2.1] bicyclic monoterpenes (borneol, bornyl acetate and camphor) and β-caryophyllene are considered to be the main aroma-active compounds of both extraction methods., (Copyright © 2014 John Wiley & Sons, Ltd.)

Over-expression of the Bcl-2 anti-apoptotic proteins is closely related to tumorigenesis and associated with drug resistance. Here we report that luteolin, a main substance found in Flos Chrysanthemi, directly binds to and shows inhibitory activity against the Bcl-2 protein. We studied the binding mode of luteolin and its derivatives with target proteins, their structure-activity relationship, and their effect on the human leukemia cell line HL-60. The results suggest that luteolin and its derivatives with a benzyl group introduced to the B ring, are new small molecule Bcl-2 protein inhibitors, and their anti-tumor activity is likely related to their effect on the Bcl-2 protein., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

White chrysanthemum flower is one of the most popular plants found everywhere in China and used as herbs. In the present work, three-dimensional fluorescence technique was used to discriminate species of white chrysanthemum flowers. Parameters affecting extraction efficiency were investigated. Under the optimal conditions, the three-dimensional fluorescence characteristics of three types of white chrysanthemum flowers were obtained. It was found that there were two main fluorescence peaks with remarkable difference in fluorescence intensity, one was corresponding to flavonoids and another was attributed to chlorophyll-like compounds. There were remarkable differences among the contours of the three white chrysanthemum flowers. Further studies showed that the fluorescence intensity ratios of chlorophyll-like compounds to flavonoids had a certain relationship with the species; those for Huai, Hang and Huangshan white chrysanthemum flowers were 6.9-7.4, 18.9-21.4 and 73.6-84.5, respectively. All of the results suggest that three-dimensional fluorescence spectra can be used for the discrimination of white chrysanthemum flowers with the advantages of low cost, ease for operation and intuition., (Copyright © 2014 Elsevier B.V. All rights reserved.)

In the present study, quantitative analysis coupled with toxic evaluation was applied to investigate the influence of sulfur-fumigation on the quality of Chrysanthemum morifolium. The results showed that the average contents of six flavonoids (cynaroside, hyperoside, isoquercitrin, luteolin, apigenin, and diosmetin) in sulfur-fumigated C. morifolium samples decreased by about 6.2%, 3.5%, 22.0%, 33.3%, 26.0%, and 36.9%, respectively, while the average contents of two organic acids (chlorogenic acid and isochlorogenic acid A) in sulfur-fumigated C. morifolium samples increased by about 16.8% and 7.6%, respectively. Furthermore, high dosage sulfur-fumigated C. morifolium could induce hydropic degeneration in epithelial cells of the renal tubule and interstitial edema. This indicates that sulfur-fumigated C. morifolium may lead to renal toxicity.

Chrysanthemum zawadskii has been proven to possess hair growth activity and has been used as treatment for hair loss. The aim of this study was to provide a novel explanation of the mechanism by which Chrysanthemum zawadskii extracts (CZe) promote hair growth and to characterize the affected hair follicle (HF) regions and the progression of growth. The n-butanol and water fractions of CZe were used for hair growth induction by topical application to the backs of C57BL/6 mice for up to 30 days. To investigate cell development during HF morphogenesis, bromodeoxyuridine-labeled skin sections were detected using immunohistochemistry. The results showed that the water fraction of CZe promoted hair shaft production and induced premature entry of telogen HFs into the anagen. Subsequently, immunohistochemical studies indicated that the water fraction of CZe stimulated the differentiation and proliferation of pluripotent epidermal matrix cells in the matrix region and epithelial stem cells in the basal layer of the epidermis. Additionally, flavonoids were identified as effective constituents. Therefore, the findings of this study suggested that the water fraction of CZe may be developed as a therapeutic agent for the prevention of hair loss.

In this study, a new method based on ultrafiltration liquid chromatography-mass spectrometry (UF-LC-MS) combined with enzyme channel blocking (ECB) was developed to discover bioactive components from herbal medicines. Xanthine oxidase (XOD), a critical enzyme for treating gout, was employed as the target protein for screening. By comparing chromatographic profiles of the compounds binding to XOD before and after the ECB experiment, the selective ligands could be distinguished from the non-selective binders. In this experiment, febuxostat bound to the channel entering into the active site of the enzyme and prevented potential ligands from binding. Finally, four compounds, namely, luteolin-7-O-glucoside, apigenin-7-O-glucoside, luteolin and apigenin were screened and identified as the candidate XOD inhibitors based on the ultrafiltration chromatogram of Flos Chrysanthemum, a famous traditional Chinese medicine used in many prescriptions for gout treatment. To verify the compounds screened further, a microplate method was applied to evaluate their enzyme inhibitory activities. The IC50 values of the above 4 compounds were 23.61, 38.80, 1.54 and 1.96μM, respectively. The structure-function relationship was also estimated according to the in vitro assay. The results were in favor of the hypothesis that the Flos Chrysanthemum extract might be used for gout treatment by inhibiting XOD., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Five flavones were isolated from Chrysanthemum coronarium L., four of which were isolated for the first time from the genus Chrysanthemum. Two were the flavonoid aglycones 5,7-dihydroxy-3,6,4'-trimethoxyflavone (1) and scutellarin-6,7-dimethyl ether (2). A new flavonoid glycoside, apigenin-7-O-[2"(6'''-O-beta-D-acetylglucopyranosyl)]-6"-O-acetylglucopyranoside (3), along with two known ones, i. e. apigenin-7-O-(2"-O-beta-D-glucopyranosyl)-beta-D-glucopyranoside (4) and 6-methoxy quercetin-7-O-beta-D-glucopyranoside (5), were identified. Structures were elucidated by NMR and MS. The therapeutic value of petroleum ether, ethyl acetate, and methanol extracts, respectively, in rats suffering from hypercholesterolemia--as a consequence of high-fat diet-and hyperglycemia--as a consequence of hypercholesterolemia and low doses of streptozotocin--was investigated through determination of biochemical markers and histopathology. The ethyl acetate and methanol extracts showed remarkable results, followed by the petroleum ether extract.

The anti-inflammatory activity of handelin (1), a guaianolide dimer from Chrysanthemum boreale flowers, was evaluated in vivo, and the effects on mediators nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) and the nuclear factor-κB (NF-κB) and ERK/JNK signaling pathways were investigated in vitro. Compound 1 inhibited lipopolysaccharide (LPS)-induced production of NO and PGE2 in cultured mouse macrophage RAW 264.7 cells. The suppression of NO and PGE2 production by 1 was correlated with the downregulation of mRNA and protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Compound 1 also suppressed the induction of pro-inflammatory cytokines TNF-α and IL-1β in LPS-stimulated RAW 264.7 cells. To further clarify the transcriptional regulatory pathway in the expression of iNOS and COX-2 by 1, the role of NF-κB was determined in RAW 264.7 cells. Compound 1 inhibits the binding activity of NF-κB into the nuclear proteins. The transcriptional activity of NF-κB stimulated with LPS was also suppressed by 1, which coincided with the inhibition of IκB degradation. Compound 1 also suppressed the activation of mitogen-activated protein kinases, including ERK and JNK signaling. In addition, the LPS-stimulated upregulation of miRNA-155 expression was suppressed by 1. The oral administration of 1 inhibited acute inflammation in carrageenan-induced paw and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced ear edema models. The serum level of IL-1β was also inhibited by 1 in a carrageenan-induced paw edema model. These findings suggest that the suppression of NF-κB activation and pro-inflammatory cytokine production may be a plausible mechanism of action for the anti-inflammatory activity of handelin.

The traditional after-harvesting drying method of C. morifolium cv. Hang-ju (HJ) is sun drying, but recently sulfur fumigation is increasingly used as a cheap and convenient method. However, the effects of sulfur fumigation on chemical constituents and potential activities of HJ were unknown. A comprehensively comparison of the chemical profiles between non-fumigated HJ (NHJ) and sulfur-fumigated HJ (SHJ) was conducted by HPLC fingerprints analysis and the discrepant peaks were identified or tentatively assigned by HPLC-ESI/MS(n). Dramatic chemical changes were found that the contents of 4 flavonoid aglycones remarkably increased while those of 7 glycosides significantly reduced which suggested that sulfur-fumigation induced flavonoid glycosides transformed into aglycons by hydrolysis reaction. A significant loss of hydroxycinnamoylquinic acids showed the sulfur fumigation was a destructive effect on HJ. Principal component analysis (PCA) was employed to rapidly discriminate NHJ and SHJ samples. By ICP-OES analysis, it was found that the residue of sulfur of SHJ were three times higher than NHJ (p<0.05). The antioxidant activity of NHJ and SHJ were evaluated by DPPH and FRAP assay, and the results showed that NHJ had much stronger antioxidant activities than SCF (p<0.05). Combining the results of chemical analysis, residue of sulfur and pharmacological evaluation, it showed that the sulfur fumigation was a destructive effect on HJ., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Eight compounds from six Chrysanthemum morifolium cv. 'Hangju' were determined and multivariate statistics, including principle component analysis (PCA) and partial least squares discriminate analysis (PLS-DA) were used to investigate the potential damaging effect of sulfr-fumigating process. Meanwhile, S, Mn, Fe, Cu, Pb were also detected by using ICP-MS and ICP-OES. In this study, dramatic chemical changes were found that the contents of flavonoid aglycones remarkably increased while those of glycosides and hydroxycinnamoylquinic acids were significantly reduced. The PCA score plots showed six samples were clearly classified into the non-fumigated and sulfur-fumigated types. And according to VIP > 1, the most important chemical markers were luteolin, apigenin and luteolin-7-O-glucoside which indicated that the extracted three components might have a marked effect on the discrimination among different group samples. And then, it was found that the residue of sulfur of SHJ were significantly higher than NHJ (P < 0.05). The established approach was applied to rapidly discriminate sulfur-fumigated HJ with combining the quantitative chemical analysis and multivariate statistical analysis, and then the result will provide some evidence to evaluat the quality of HJ and control its processing.

Investigations of the transfer of pesticide residues from tea to its infusion can be important in the assessment of the possible health benefits of tea consumption. In this work the transfer of difenoconazole and azoxystrobin residues from chrysanthemum tea to its infusion was investigated at different water temperatures, infusion intervals and times. The transfer percentages were in the range of 18.7-51.6% for difenoconazole and of 38.1-71.2% for azoxystrobin, and increased considerably with longer infusion intervals. The results indicated that azoxystrobin with a lower octanol-water partition coefficient of 2.5, showed a higher transfer than that of difenoconazole with a relatively high octanol-water partition coefficient of 4.4. Water temperature had no significant effect on the transfer of the two residues, and no obvious loss of difenoconazole and azoxystrobin occurred during the infusion process. The concentrations in the infusions decreased gradually from 0.67 to 0.30 μg kg(-1) for difenoconazole and from 2.3 to 0.46 μg kg(-1) for azoxystrobin after five infusions. To assess the potential health risk, the values of estimate expose risk were calculated to be 0.016 for difenoconazole and 0.0022 for azoxystrobin, meaning the daily residue intake of the two analytes from chrysanthemum tea was safe. This research may help assure food safety and identify the potential exposure risks from pesticides in chrysanthemum that may be health concerns.

A water-soluble polysaccharide (P2) with a molecular weight of 1.7×10(5)Da was isolated from the hot aqueous extract of flowers of the Chrysanthemum morifolium. It was homogeneous for there was only a symmetrical peak on the spectrum of high-performance gel permeation chromatography. Monosaccharide analysis revealed that P2 is an arabinogalactan containing arabinose (38.4% w/w), galactose (58.8% w/w), and glucose (2.8% w/w) in a ratio of 1:1.53:0.07. The GC-MS results of the derived alditol acetates from the permethylated P2 showed some separate peaks corresponding to (1→6)-linked Gal, (1→3,6)-linked Gal, terminal Ara, (1→5)-linked Ara, and (1→3,5)-linked Ara units in a molar ratio of nearly 1.12:1.90:1.84:0.35:0.12. Terminal Glc unit also existed in a trace amount. (1)H, (13)C, DEPT-135 and 2D NMR (including HMQC and HMBC), as well as graded acid hydrolysis experiment, indicated that the core structure features include a backbone chain consisting of (1→6)-linked and (1→3,6)-linked Galp residues. Side chains which were composed of terminal Glcp, terminal, (1→5)-linked and (1→3,5)-linked Araf residues were located at position 3. Additionally, P2 showed good immunological activity as it significantly enhanced splenocyte proliferation in relatively low 25 and 50μg/mL doses. However, P2 showed little effects of thymocyte proliferation, indicating that it may have two modes of action., (Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.)

Some polyphenols derived from plants may ameliorate hyperlipidemic fatty livers; therefore, we hypothesized that polyphenol-rich Chrysanthemum morifolium extract (CME) may exert an inhibitory effect on the formation of hyperlipidemic fatty livers in mice. This study aimed to examine the effects of CME on lipids in blood and liver and on peroxisome proliferator-activated receptor (PPAR)α-mediated gene expression. Mice with hyperlipidemic fatty livers induced by orally administering high-fat milk via gavage and being simultaneously treated with 75 to 300 mg/kg CME for 6 weeks. After CME addition, the serum total cholesterol levels and hepatic weight coefficients decreased, but no significant reduction in the serum triacylglycerol levels were observed. It is important to note that CME might decrease hepatic lipid accumulation, sterol regulatory element binding protein-1c, and fatty acid synthase expression and increase hepatic PPARα, lipoprotein lipase, and cholesterol 7α-hydroxylase expression. However, the expected reduction in hepatic diacylglycerol acyltransferase mRNA expression was not observed. These findings demonstrate that polyphenol-rich CME may prevent hyperlipidemic fatty liver in mice, and its mechanisms may be related to the modulation of sterol regulatory element binding protein-1c, FAS, lipoprotein lipase, and cholesterol 7α-hydroxylase 1 expression through the PPARα-mediated pathway., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The interaction of stem cell factor (SCF) with its cognate receptor c-Kit is closely associated with the survival and maturation of melanocytes. To investigate novel depigmentation agents, we screened 2,000 plant extracts for c-Kit inhibitors to identify active small molecules by using time-resolved fluorescence enzyme assays. For the active extracts identified as inhibitors of c-Kit enzyme, we evaluated the effects of the active extracts and isolated flavonoids on c-Kit phosphorylation in MO7e/melanocytes. Anti-melanogenic activity was also examined in melanocytes and melanoderm model. The flavonoids such as diosmetin, apigenin, acacetin and luteolin isolated from Chrysanthemum morifolium were found to be active in inhibiting c-Kit both at enzyme and cellular levels. In addition, these flavonoids attenuated SCF-induced proliferation of human primary melanocytes without toxicity and suppressed ultraviolet (UV) B irradiation-mediated melanin synthesis significantly. Among the active flavonoids, diosmetin was found to inhibit SCF-induced melanogenesis in a human melanoderm model. These results strongly suggest that C. morifolium extract and diosmetin have potential to suppress SCF-/UVB-induced melanogenesis, and could be developed as anti-pigmentation agents.

Tomato (Solanum lycopersicum) crops can be severely damaged due to parasitism by the root-knot nematode (RKN) Meloidogyne incognita, but are protected when intercropped with crown daisy (Chrysanthemum coronarium L.). Root exudate may be the determining factor for this protection. An experiment using pots linked by a tube and Petri dish experiments were undertaken to confirm that tomato-crown daisy intercropping root exudate decreased the number of nematodes and alleviated nematode damage, and to determine crown daisy root exudate-regulated nematode chemotaxis. Following a gas chromatography-mass spectrometry assay, it was found that the intercropping protection was derived from the potent bioactivity of a specific root exudate component of crown daisy, namely lauric acid. The Mi-flp-18 gene, encoding an FMRFamide-like peptide neuromodulator, regulated nematode chemotaxis and infection by RNA interference. Moreover, it was shown that lauric acid acts as both a lethal trap and a repellent for M. incognita by specifically regulating Mi-flp-18 expression in a concentration-dependent manner. Low concentrations of lauric acid (0.5-2.0mM) attract M. incognita and consequently cause death, while high concentrations (4.0mM) repel M. incognita. This study elucidates how lauric acid in crown daisy root exudate regulates nematode chemotaxis and disrupts Mi-flp-18 expression to alleviate nematode damage, and presents a general methodology for studying signalling systems affected by plant root exudates in the rhizosphere. This could lead to the development of economical and feasible strategies for controlling plant-parasitic nematodes, and provide an alternative to the use of pesticides in farming systems.

The supercritical-carbon dioxide fluid extract of Chrysanthemum indicum Linné. (CFE) has been demonstrated to be effective in suppressing inflammation. The aim of this study is to investigate the preventive action and underlying mechanisms of CFE on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in mice. ALI was induced by intratracheal instillation of LPS into lung, and dexamethasone was used as a positive control. Results revealed that pretreatment with CFE abated LPS-induced lung histopathologic changes, reduced the wet/dry ratio and proinflammatory cytokines productions (TNF-α, IL-1β, and IL-6), inhibited inflammatory cells migrations and protein leakages, suppressed the levels of MPO and MDA, and upregulated the abilities of antioxidative enzymes (SOD, CAT, and GPx). Furthermore, the pretreatment with CFE downregulated the activations of NF-κB and the expressions of TLR4/MyD88. These results suggested that CFE exerted potential protective effects against LPS-induced ALI in mice and was a potential therapeutic drug for ALI. Its mechanisms were at least partially associated with the modulations of TLR4 signaling pathways.

Airborne dermatitis belongs to a heterogeneous group of dermatoses of various etiopathology and clinical characteristics. This disease is characterized by acute or chronic inflammation of the uncovered skin exposed to irritants or allergens. Initially skin lesions are transient. The paper presents a description of chrysanthemum growers diagnosed with air-borne dermatitis from chrysanthemum. Etiology, pathomechanism, clinical course, diagnostics and therapeutical methods are described.

The present work reports a simple, cost-effective, and ecofriendly method for the synthesis of silver nanoparticles (AgNPs) using Chrysanthemum indicum and its antibacterial and cytotoxic effects. The formation of AgNPs was confirmed by color change, and it was further characterized by ultraviolet-visible spectroscopy (435 nm). The phytochemical screening of C. indicum revealed the presence of flavonoids, terpenoids, and glycosides, suggesting that these compounds act as reducing and stabilizing agents. The crystalline nature of the synthesized particles was confirmed by X-ray diffraction, as they exhibited face-centered cubic symmetry. The size and morphology of the particles were characterized by transmission electron microscopy, which showed spherical shapes and sizes that ranged between 37.71-71.99 nm. Energy-dispersive X-ray spectroscopy documented the presence of silver. The antimicrobial effect of the synthesized AgNPs revealed a significant effect against the bacteria Klebsiella pneumonia, Escherichia coli, and Pseudomonas aeruginosa. Additionally, cytotoxic assays showed no toxicity of AgNPs toward 3T3 mouse embryo fibroblast cells (25 μg/mL); hence, these particles were safe to use.

We applied Chrysanthemum flower oil (CFO) to a hyperuricemia model by feeding rats a hyperuricemia-inducing diet (HID) and investigated its effect on serum uric acid (SUA) levels and its mode of action. CFO is the oily fraction that contains polyphenols derived from chrysanthemum flowers. Oral administration of CFO to HID-fed rats significantly decreased their SUA levels. It also inhibited xanthine oxidase activities in the liver and increased urine uric acid levels. The effects of CFO on the renal gene expressions that accompanied the induction of hyperuricemia were comprehensively confirmed by DNA microarray analysis. The analysis showed up-regulation of those genes for uric acid excretion by CFO administration. These results suggest that CFO suppresses the increase in SUA levels via two mechanisms: suppression of uric acid production by inhibition of xanthine oxidase in the liver and acceleration of its excretion by up-regulation of uric acid transporter genes in the kidney.

A new rapid UHPLC-UV-QTOF/MS method has been developed for the simultaneous analysis of nine phenolic compounds [(Z)-2-β-d-glucopyranosyloxy-4-methoxycinnamic acid (cis-GMCA), chlorogenic acid, (E)-2-β-d-glucopyranosyloxy-4-methoxycinnamic acid (trans-GMCA), quercetagetin-7-O-β-d-glucopyranoside, luteolin-7-O-β-d-glucoside, apigenin-7-O-β-d-glucoside, chamaemeloside, apigenin 7-O-(6″-O-acetyl-β-d-glucopyranoside), apigenin] and one polyacetylene (tonghaosu) from the flower heads of Chamomile/Chrysanthemum samples. The chromatographic separation was achieved using a reversed phase C18 column with a mobile phase of water and acetonitrile, both containing 0.05% formic acid. The ten compounds were completely separated within 15min at a flow rate of 0.25mL/min with a 2μL injection volume. The different chemo-types of Chamomiles/Chrysanthemum displayed variations in the presence of chemical constituents. German Chamomile samples confirmed the presence of cis-GMCA, trans-GMCA, apigenin-7-O-β-d-glucoside and tonghaosu as major constituents whereas Roman chamomile samples confirmed the presence of chamamaeloside and apigenin as major compounds. The Chrysanthemum morifolium samples showed the presence of luteolin-7-O-β-d-glucose as the major compound. The method was applied for the analysis of various commercial products including capsules, tea bags, body and hair care products. LC-mass spectrometry with electrospray ionization (ESI) interface method is described for the evaluation of ten compounds in plant samples and commercial products. This method involved the detection of [M+Na](+) and [M+H](+) ions in the positive mode. Partial least squares discriminant analysis (PLS-DA) was used to visualize commercial samples quality and may be of value for discriminating between chamomile types and Chrysanthemum with regards to the relative content of individual constituents. The results indicated that the method is suitable as a quality control test for various Chamomile/Chrysanthemum samples and market products., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The essential oils from the leaves, stems and roots of Chrysanthemum trifurcatum (Desf.) Batt. and Trab. var. macrocephalum (viv.) were obtained by hydrodistillation and their chemical compositions were analysed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS), in order to get insight into similarities and differences as to their active composition. A total of fifty compounds were identified, constituting 97.84%, 99.02% and 98.20% of total oil composition of the leaves, stems and roots, respectively. Monoterpene hydrocarbons were shown to be the main group of constituents of the leaves and stems parts in the ratio of 67.88% and 51.29%, respectively. But, the major group in the roots oil was found to be sesquiterpene hydrocarbons (70.30%). The main compounds in leaves oil were limonene (26.83%), γ-terpinene (19.68%), α-pinene (9.7%) and α-terpenyl acetate (7.16%). The stems oil, contains mainly limonene (32.91%), 4-terpenyl acetate (16.33%) and γ-terpinene (5.93%), whereas the main compounds in roots oil were α-calacorene (25.98%), α-cedrene (16.55%), β-bourbobene (14.91%), elemol (7.45%) and 2-hexenal (6.88%). The crude organic extracts of leaves, stems and roots, obtained by maceration with solvents of increasing polarity: petroleum ether, ethyl acetate and methanol, contained tannins, flavonoids and alkaloids. Meanwhile, essential oils and organic extracts were tested for antibacterial activities against eight Gram-positive and Gram-negative strains, using a microdilution method. The oil and methanolic extact from C. trifurcatum leaves showed a great potential of antibacterial effect against Bacillus subtilis and Staphylococcus epidermidis, with an IC50 range of 31.25-62.5 µg/ml.

Chrysanthemum is a ubiquitous plant with many species and wide uses, and it is usually consumed as functional food. The main aim of this paper is to demonstrate that chromatographic fingerprints obtained from the HPLC/UV analysis of the pressurized hot water extraction (PHWE) extracts together with the aid of principal component analysis (PCA), allowed for the clustering of various chrysanthemums of different species and provenance. In addition, a parallel study of pressurized fluid extraction (PFE) with methanol was carried out for comparison. From the results, a clearer separation and clustering was obtained with the environmentally-benign water extracts compared with methanol extracts. This study shows that PHWE in combination with HPLC/UV and PCA can be used successfully as a green and effective approach for characterisation and quality control of ubiquitous functional food such as chrysanthemum., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

In this study, we report the anti-proliferative effect and molecular mechanism of Chrysanthemum indicum (C. indicum) on A549 human alveolar basal epithelial cells. We also analyzed the changes in C. indicum component profiles due to modifications of predrying process, flower size, and extraction method. Among the varieties of modifications tested, high-temperature heat dry (HTD) of small flower biotype followed by the methanolic extraction resulted in the strongest anti-proliferative activity of C. indicum extract in A549 cells. High-performance liquid chromatography of C. indicum revealed that the levels of acacetin 7-O-rutinoside (linarin) are markedly increased by heat treatment, especially HTD. Finally, we showed that linarin-mediated inhibition of cell proliferation is associated with suppression of Akt activation and induction of cyclin-dependent kinase inhibitor p27(Kip1) as evidenced by cell cycle analysis and treatment with LY294002, an inhibitor of phosphatidylinositol 3-kinase/Akt pathway. Taken together, these findings suggest the need for further development and evaluation of linarin from C. indicum for the treatment and prevention of lung cancer.

We have previously reported that seventy percent ethanol extract of Chrysanthemum indicum Linne (CIE) strongly reduces Epstein-Barr virus (EBV)-transformed lymphoblastoid cell line (LCL) survival by inhibiting virus-encoded latent infection membrane protein 1 (LMP1)-induced NF-κB activation. To identify an active compound(s) in CIE that inhibits LMP1-induced NF-κB activation, activity-guided fractionation was employed. The CH2Cl2 fraction of CIE strongly reduced LMP1-induced NF-κB activation and LCL viability with relatively low cytotoxic effects on primary human foreskin fibroblast (HFF), HeLa or Burkitt's lymphoma (BL41) cells. Furthermore, lupeol, a pentacyclic triterpene, was identified in the CH2Cl2 fraction of CIE to attenuate LMP1-induced NF-κB activation and LCL viability. This study demonstrates that lupeol is one of active compounds in the CH2Cl2 fraction of CIE that inhibits LMP1-induced NF-κB activation and reduces NF-κB-dependent LCL viability.

Objective: To study the effects of total flavones of Chrysanthemum indicum on proliferation and apoptosis of human osteosarcoma Saos-2 cells and its mechanism., Methods: The effect of the total flavones of Chrysanthemum indicum on the proliferation of human osteosarcoma Saos-2 cells was detected by CCK assay, and the morphological changes of cells treated with total flavones of Chrysanthemum indicum were observed using contrast microscope. Flow cytomerty was performed to analyze the apoptotic rate of the cells, and the gene expression levels of Caspase-3, BCL-2, BAX were detected by RT-PCR., Results: The total flavones of Chrysanthemum indicum suppressed the proliferation of osteosarcoma cells in a dose-and time-dependent manner. Under a microscope observation of cell morphology, the volume became smaller ,the number of internal particles was increased. Cell apoptosis rate was positively related to the drug concentration. After treated for 48 hours, Caspase-3 and BAX expression were up-regulated, BCL-2 and BCL-2/BAX were decreased., Conclusion: The total flavones of Chrysanthemum indicum can inhibit the proliferation of osteosarcoma cell line Saos-2 by inducing cell apoptosis,the mechanism of which might be related with reducing BCL-2/BAX and activating Caspase-3.

The contents of chlorogenic acid, 3,5-O-caffeoylquinic acid, galuteolin and quercitrin in Sanvitalia procumbens and Chrysanthemum morifolium cv. 'Hangju' and 'Gongju' were determined by RP-HPLC. The main active ingredient of S. procumbens was similar to C. morifolium cv. 'Hangju' and 'Gongju'. The content varied significantly. The contents of chlorogenic acid, quercitrin and galuteolin in S. procumbens were 7.46, 46.58, 26.01 mg x g(-1), respectively, and they were the highest among the samples. The content of 3,5-O-caffeoylquinic acid in C. morifolium cv. 'Hangju' was 14.70 mg x g(-1), it was the highest among the samples. The data of the study provide a basis for further research and development of S. procumbens.

Unlabelled: To observe and analyze the character change of Chinese Chrysanthemi Flos from different producing areas in the same storage conditions, and provide certain reference to the study of storage method and storage time of Chrysanthemi Flos., Method: Different species of Chrysanthemi Flos were collected from the whole country at the same period, stored in the same nature conditions for three years. The change on worm situation, mildew degree, flower shape, color and appearance character were ovserved., Result: The degree of worm and mildew situation of different regions of Chrysanthemi Flos was: Qiju < Hangju < Gongju < Chuju < Jiju = Huaiju = Damaya chrysanthemum. Among three cultivar species of Hangju, 'Huju' was the best preserved, following by 'Xiaobaiju', the worst was 'Dabaiju'. After the storage, the degree of worm situation of Gongju was different. The degree of worm situation of Chuju and Qiju were related to the process methods., Conclusion: The storage changes of Chrysanthemi Flos were related to the regions, specise, process methods, geography location and harvest time, etc. Different specises of Chrysanthemi Flos had different storage time.

The Chrysanthemum lavandulifolium extract, which includes chrysoeriol, sudachitin, and acacetin, has excellent antibiotic effects on Escherichia coli O157:H7 (E. coli O157). A notable point is that the antibiotic targets of the herb extract are similar to the targets of commonly used antibiotic drugs, including bacterial cell wall biosynthesis, bacterial protein synthesis, and bacterial DNA replication and repair. In addition, the herbal antibiotic inhibits the etiological factors that contribute to the pathogenic property. The herbal sample was extracted and fractionated and then inoculated through a disk diffusion method to confirm its antibiotic effect against E. coli O157. Total RNA was isolated from the affected bacterial cells, and its expression level was analyzed through a microarray analysis. To confirm the accuracy of the microarray data, a real-time PCR was performed. Three active compounds, chrysoeriol, sudachitin, and acacetin, were identified with a high-performance liquid chromatography-electrospray ionization/mass spectrometry chromatogram, and the disk diffusion study confirmed that chrysoeriol and sudachitin contribute to the antibiotic properties of the herb extract. The results demonstrate that the multi-target efficacy of the herbal sample may indicate the potential for the development of more effective and safer drugs that will act as substitutes for existing antibiotics., (Copyright © 2012 John Wiley & Sons, Ltd.)

In this study, Chrysanthemum zawadskii extract (CZE) was investigated to determine its effects on 2-deoxy-D-ribose (dRib)-induced oxidative damage and cellular dysfunction in the MC3T3-E1 mouse osteoblastic cell line. Osteoblastic cells were treated with the highly reducing sugar, dRib, in the presence or absence of CZE. Cell viability, apoptosis and reactive oxygen species (ROS) production were subsequently examined. It was observed that dRib reduced cell survival, while it markedly increased the intracellular levels of ROS and apoptosis. However, pre-treatment of the cells with CZE attenuated all the dRib-induced effects. The antioxidant, N-acetyl-L-cysteine (NAC), also prevented dRib-induced oxidative cell damage. In addition, treatment with CZE resulted in a significant increase in alkaline phosphatase (ALP) activity and collagen content, as well as in the expression of genes associated with osteoblast differentiation [ALP, collagen, osteopontin (OPN), osteoprotegerin (OPG), bone sialoprotein (BSP), osteocalcin (OC) and bone morphogenetic protein (BMP)2, BMP4 and BMP7]. In mechanistic studies of the antioxidative potential of CZE, we found that CZE reversed the dRib-induced decrease in the expression of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT)1 and AKT2 genes, which are master regulators of survival-related signaling pathways. CZE also upregulated the gene expression of the antioxidant enzymes, superoxide dismutase (SOD)2, SOD3 and glutathione peroxidase 4 (GPx4), which was inhibited by dRib. Taken together, these results suggest that CZE attenuates dRib-induced cell damage in osteoblastic cells and may be useful for the treatment of diabetes-associated bone disease.

This study is aimed to establish a high-performance liquid chromatography (HPLC) method for simultaneous determination of chlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, luteolin-7-O-beta-D-glucoside, 3,4-dicaffeoylquinic acid, linarin and luteolin in Chrysanthemum indicum. The separation was carried out on a Shim pack VP-ODS (4.6 mm x 250 mm, 5 microm) column eluting with mobile phases of methanol (A) and water containing 0.3% phosphoric acid (B) in gradient mode (0-9 min, 85% -80% B; 9-12 min, 80% -70% B; 12-15 min, 70% -65% B; 15-20 min, 65% -60% B; 20-23 min, 60% -55% B; 23-29 min, 55% -54.4% B; 29-32 min, 54.4% -45% B; 32-37 min, 45% -5% B; 37-45 min, 5% -85% B) at the flow rate of 1.0 mL x min(-1). The column temperature was 35 degrees C and the detection wavelength was set at 326 nm. The good separation of chlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, luteolin-7-O-beta-D-glucoside, 3,4-dicaffeoylquinic acid, linarin and luteolin was achieved within 40 min. Calibration curves of the eight effective components showed good linear relationship (r > 0.999 5, n = 7). The average recoveries were within 97.03%-102.3% (RSD < 2.0%, n = 6). The method is simple, accurate and repeatable and can be used for the quality control of Ch. indicum.

The widespread use of insecticides in Chinese herbal medicines has created a compelling need for the development of a multiresidue analytical method to help assure herbs safety. The operating variables affecting the performance of the multiresidue analysis of 34 organochlorine and 12 pyrethroid pesticides in chrysanthemum, a widely used kind of Chinese herbal medicines, were evaluated. Three different extraction solvents including n-hexane and its mixtures with acetone and petroleum ether were compared, and n-hexane was found to be an appropriate option. A combination of gel permeation chromatography and SPE was selected as the optimum cleanup, in comparison with dispersive SPE, or the two former methods alone. The determination of the 46 pesticide residues in the spiked chrysanthemum samples was performed by GC with electron capture detection. The average recoveries ranged from 71.3 to 102.6% with RSDs of 1.4-15.7% for all of the pesticides. The LOQs were in the range of 0.0015-0.2 mg/kg, while the LODs were between 0.0005 and 0.1 mg/kg. The satisfactory accuracy, and precision, in combination with a good separation and few interferences, have demonstrated the strong potential of this technique for its application in chrysanthemum analysis., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objective: To explore quantitative assessment indicators of Chrysanthemi Flos, and optimize the extraction process of Chrysanthemi Flos through orthogonal experimental design., Method: The concentration of ethanol, amount of ethanol, extraction time and extraction frequency were selected as factors in the L9 (3(4)) orthogonal experiment. A comprehensive assessment was conducted with the peak area of the eight major common peaks in the fingerprint of Chrysanthemi Flos as the indicators., Result: The optimum extraction process was selected as follows: using ultrasonic extraction method, adding 30-fold ethanol with 80% concentration, extracting for 2 times for extraction, 40 min for each time., Conclusion: The optimized extraction process is reliable, with controllable assessment indicators, which is significant to the standardization of the extraction process and quality control of Chrysanthemi Flos preparations.

The aims of the present study were to study the interspecies difference in the pharmacokinetics of luteolin and apigenin occurring in Chrysanthemum morifolium extract (CME) among rats, beagle dogs, mini-pigs, and humans, and compared the human pharmacokinetic parameters with the data predicted from the above three animals. The plasma concentrations of luteolin and apigenin were determined with a RP-HPLC method. An interspecies difference of pharmacokinetics was found, especially between rats and other species, the plasma concentration of luteolin was much lower than that of apigenin in rats, although the content of luteolin in CME was higherthan that of apigenin, whereas the plasma concentration of luteolin was much higher than that of apigenin in dogs, mini-pigs and humans. Animal scale-up of some pharmacokinetic parameters of luteolin and apigenin were also performed after rats, beagle dogs, mini-pigs and humans were orally given CME at dosages of 400 mg/kg, 102 mg/kg, 90 mg/kg, and 20 mg/kg, respectively. Linear relationships were obtained between log mean retention time (MRT) and log species body weight (W) (kg), and log elimination half-life (t1/2) (h) and logW. The corresponding allometric equations were MRT=9.382W(0.1711) (R2 = 0.9999) and t1/2 = 4.811W(0.1093) (R2 = 0.9013) for luteolin, MRT = 12.53W(0.0356) (R2 = 0.9980) and t1/2 = 7.940W(0.0294) (R2 = 0.9258) for apigenin, respectively. The predicted human pharmacokinetic parameters (MRT and t1/2) by an allometric approach were 18.6 h and 7.46 h for luteolin, 14.3 h and 8.95 h for apigenin, respectively, which were close to the values obtained from humans (20 mg CME/kg) in the present study. The study has demonstrated the possibility to extrapolate the pharmacokinetic behavior of flavonoids from animals to humans.

Objective: To establish a method for determination of trace elements in chrysanthemum and analyze trace elements in chrysanthemum from different habitats., Methods: The preprocessing for determination of trace elements in chrysanthemum was carried out by microwave digestion,the content of trace elements in chrysanthemum was determined by ICP-MS., Results: The content of trace elements of chryscmthemum varied with the habitats., Conclusion: The contents of trace elements as well as heavy metals of Chrysanthemum relate to its species and habitats to a certain degree.