Flos Chrysanthemi Indici Research Articles

The screening and evaluation of herbs and identification of herbal combinations with anti-viral effects on Newcastle disease virus

A total of 25 “heat-clearing and detoxifying” herbs used in Chinese medicine were investigated for their cytopathic effects on the growth of Newcastle Disease virus (NDV) in a chicken fibroblast cell line.The 5 herbs with the highest virus inhibitory effects were Herba agastaches, Flos chrysanthemi indici, Rhizoma anemarrhenae, Astragalus root and Baikal skullcap root and these were used in herbal formulations. Anti-NDV activities of 4 formulations were tested on the growth of NDV in the DF-1 fibroblast cell line.Formulation II, containing Baikal skullcap root, Astragalus root, Anemarrhena rhizome (1:1:2) and formulation IV containing Anemarrhena rhizome, Astragalus root and Flos chrysanthemi indici (1:1:1), which had strong anti-NDV activity in vitro, were used to determine the in vivo inhibitory effects of NDV-infection in chickens. After treatment with the two formulations serum IgY titres against NDV were improved, and morbidity was reduced in the NDV-infected chickens.The results suggest that the components in formulations II and IV acted synergistically to improve resistance to Newcastle disease and provide a basis for the developing an anti-NDV herbal medicine.

Preparative Purification of Bioactive Compounds from Flos Chrysanthemi Indici and Evaluation of Its Antiosteoporosis Effect.

To understand the material basis and underlying molecular machinery of antiosteoporosis activity of the Flos Chrysanthemi Indici (FCI), the consequences of ethanol extract on the bone loss in mice induced due to ovariectomy (OVX) was evaluated. Also, the antiosteoporosis fraction obtained from the FCI ethanol extract was isolated and purified using a preparative high-speed countercurrent chromatography (HSCCC). The in vitro impact of the compounds was investigated on osteoblast proliferation and differentiation. The results revealed that ethyl acetate fraction with robust in vivo antiosteoporosis activity was obtained. The important compounds purified by HSCCC using gradient elution system included acacetin, apigenin, luteolin, and linarin. The four compounds enhanced the differentiation and proliferation of osteoblasts in MC3T3-E1 cells. They also augmented the mRNA levels of runt-related transcription factor 2 (Runx2), osteocalcin (OCN), osteopontin (OPN), and type I collagen (COL I). The AKT signaling pathway was also activated in MC3T3-E1 cells by the four compounds. The present study demonstrated that the antiosteoporosis effects of FCI did not depend on a single component, and HSCCC efficiently isolated and purified the antiosteoporosis bioactive compounds from FCI.

The analyses and identification of Flos rhododendri mollis and Flos chrysanthemi indici via infrared spectroscopy

In this study, major chemical components of Flos rhododendri mollis and Flos chrysanthemi indici were characterized using Fourier transform infrared spectroscopy (FTIR). For Flos rhododendri mollis, the bands at 1,648 and 1,543 cm(-1) were attributed to amide I and amide II , respectively, indicating that it contained proteins probably resulting in immunization. In case of Flos chrysanthemi indici, stretching vibration of C==O function group was responsible for the bands at 1,734 and 1,515 cm(-1), as a result of essential oils, lipids, etc. Since FTIR spectra of Flos rhododendri mollis and Flos chrysanthemi indici are almost identical and it is difficult to discriminate them, two-step identification was investigated via secondary derivative of the FTIR spectra. The bands at 1,656 and 1,515 cm(-1) corresponds to flavonoides in Flos rhododendri mollis and Flos chrysanthemi indici. In the secondary derivative of the FTIR spectrum of Flos chrysanthemi indici, characteristic bands of inulin were present at 1,163, 1,077, 1,026, 986 and 869 cm(-1), and therefore Flos chrysanthemi indici contained inulin as well. Tri-step identification was carried out for Flos rhododendri mollis and Flos chrysanthemi indici by means of comparing their 2D-IR correlation spectra in different wave number range. In the characteristic range of flavonoides (1,700-1,400 cm(-1)), Flos rhododendri mollis exhibited 3 obvious autopeaks, while 10 autopeaks were visualized in the 2D-IR correlation spectrum of Flos chrysanthemi indici Moreover, in the characteristic range of glucoside (1,250-900 cm(-1)), 10 and 9 autopeaks were present in the 2D-IR correlation spectra of Flos rhododendri mollis and Flos chrysanthemi indici, respectively. Therefore, the tri-step identification of FTIR is a time-saving; accurate, cost-saving and convenient method to effectively distinguish traditional Chinese medicines.

Flos Chrysanthemi Indici protects against hydroxyl-induced damages to DNA and MSCs via antioxidant mechanism

Flos Chrysanthemi Indici (FCI) is a Chinese herbal medicine used in China for over 2000years. In this study, its ethanol extract (EFCL) was found to protect against hydroxyl radical (OH) induced oxidative damages to mesenchymal stem cells (MSCs) and DNA. To explore the mechanism, EFCL was further determined by chemical and antioxidant assays. Folin–Ciocalteu colorimetric assay suggested that content of total phenolics was 93.85±3.56mg catechin/g and HPLC analysis indicated a content of 10.05±1.44mg/g as chlorogenic acid in EFCI. Antioxidant assays revealed that EFCI could also scavenge O2− radical (IC50 58.74±1.30μg/mL), DPPH radical (IC50 46.46±1.35μg/mL) and ABTS+ radical (IC50 20.59±0.52μg/mL), bind Fe2+ (IC50 364.27±19.57μg/mL) and Cu2+ (IC50 734.77±34.69μg/mL), reduce Fe3+ (IC50 147.11±11.09μg/mL) and Cu2+ (IC50 35.69±3.14μg/mL). On the basis of mechanistic analysis, we concluded that FCI can effectively protect against hydroxyl-induced damages to DNA and MSCs, therefore FCI shows promise as a possible therapeutic reagent for many diseases. The effect may be mainly attributed to phenolics especially chlorogenic acid, which exerts the antioxidant action possibly through metal-chelating, and radical-scavenging which is via hydrogen atom transfer (HAT) and/or sequential electron proton transfer (SEPT) mechanisms.

English

JiMaiTong (JMT), a Chinese herb compound prescription consisted of Flos chrysanthemi Indici, Spica prunellae and Semen cassiae for anti-hypertension. Luteolin is one of the major bioactivity compositions in F. chrysanthemi Indici in JMT. There are some reports about pharmacokinetics of luteolin in extract of F. chrysanthemi and husks of peanut in normal rats, but it lacked pharmacokinetic information of luteolin residing in a Chinese herb compound prescription in hypertensive animal models. The present study aimed to develop a high-performance liquid chromatography with photodiode array detection (HPLC-DAD) method for determination of luteolin in rat plasma and for pharmacokinetic study after oral administration of JMT to spontaneous hypertensive rats (SHR) and normal Sprague Dawley (SD) rats. After oral administration of JMT to SHR and SD rats, respectively the content of luteolin in blood samples at different time points were determined by a reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with liquid-liquid phase extraction. This revealed a difference in disposition of luteolin in SHR and SD rats and a relationship between plasma concentration and hypertension of pathological state within the organism. This was the first report on the pharmacokinetic compare of luteolin between normal and hypertension pathological state rats. The hypertension pathological state would effects the disposition of luteolin, and it may provide a meaningful basis for the clinical application of JMT. Key words: Luteolin, pharmacokinetic, Flos chrysanthemi Indici, hypertension, Chinese herb.

The study of anti-inflammatory and anti-oxidant effects of the five edible plants

The anti-inflammatory and anti-oxidant effects of the five edible plants (floral herbal tea, FHT) commonly utilized as a drink in Guangdong of China were discussed. Firstly, DPPH and ABTS methods were used to determine antioxidant activity of methanolic extract from floral herbal tea (MEFH). Subsequently, the relationship between total content of phenol and flavonoids with respect to antioxidant activity were studied. Finally, anti-inflammatory activity of MEFH in RAW 264.7 cell line was tested. The scavenging activity of DPPH radicals in the order from highest to lowest was: Flos sophorae, Lonicera japonica Thunb, Flos chrysanthemi Indici, Flos puerariae, Bombax ceiba. F. Sophorae not only possessed the highest anti-oxidant activity, but also the high contents of total phenol and flavonoids. NO induced by lipopolysaccharide (LPS) in mouse macrophages can be effectively inhibited by MEFH, in which the anti-inflammatory effect of F. Sophorae was highest. Amongst five edible plants, F. sophorae demonstrated extraordinary anti-inflammatory as well as anti-oxidant effects. These effects were mainly due to high content of total phenol and flavonoids, which could serve as a natural resource of food and medicine. Key words: Anti-inflammatory, anti-oxidant, floral herbal tea, Flos sophorae.

Toxicological evaluation of carotenoid-type extracts from Flos Chrysanthemi Indici

Flos Chrysanthemi Indici (FCI) was considered as a traditional medicine and regarded as a conventional food in China, Korea and Japan. Toxicological evaluation of carotenoid-type extracts derived from FCI was investigated in the present paper. Acute toxicity of the extracts was evaluated with a single dose administered by the intragastric route to SD rats in dosages of 15 g/kg. After an observation period of 2 weeks after intragastric administration, no mortality or any changes in appearance, behavior occurred for both female and male rats in the high dosages mentioned. Macroscopic and microscopic studies of the internal organs revealed no pathological changes. The observations of acute toxicity test indicated that the extracts were nontoxic. Genetic toxicity of the extracts was assessed by Ames assay in Salmonella typhimurium strains (TA97, TA98, TA100 and TA102) with and without metabolic activation (S9), by bone marrow micronucleus test and sperm abnormality test in ICR mice. The examinations of genotoxicity assay indicated that the extracts lack genotoxic potential. Although these relatively short-term studies document no toxicity, longer-term use could result in serious toxicity. Further studies are therefore required to study long-term toxicity. Key words: Acute oral toxicity, carotenoid-type extracts, Flos Chrysanthemi Indici, genetic toxicity.

Principal component analysis and correlation analysis of elements in Flos Chrysanthemi Indici from different regions

To explore the character of inorganic elements in Flos Chrysanthemi indici and look for relationship between the element concentration and regions. The contents of elements, including borum (B), sodium (Na), magnesium (Mg), phosphorus (P), potassium (K), calcium(Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), strontium (Sr), cesium (Se), barium(Ba) and lead(Pb) in Chinese traditional herb Flos Chrysanthemi indici from different regions were determined by ICP-AES. The element distrubution diagram were plotted. The principal component analysis and correlation analysis of SPSS were applied for the study of characteristic elements. Similar curves of element concentration have been acquired. It is observed that the content of elements in the samples shows regional diversity. There are 15 correlative element pairs in correlation analysis. Four principal components which accounted for over 84.437% of the total variance were extracted from the original data. The first and second factors accounted for 60.090% of the total variance, which means that P, K, Ca, Mn, Fe, Cu, Sr, B, Na and Se may be the characteristic elements. The showed that element content in Flos Chrysanthemi Indici display special distributing diagram. Remarkable correlation is presented in some element pairs. The elements contents of Flos Chrysanthemi indici gained from Yunan, Hunan are higher than those from other regions.

Determination of chlorogenic acid, caffeic acid and linarin in Flos Chrysanthemi Indici from different places by RP-HPLC

To evaluate the quality of Flos Chrysanthemi Indici which produced in twenty-two different producing places. Chlorogenic acid and caffeic acid were analyzed on a Shim-pack C8 colunm (4.6 mm x 250 mm, 5 microm) eluted with the mobile phase consisted of acetonitrile-0.5% phosphoric acid( 19:81). The detection wavelength was set at 326 nm. Linarin were eluted with the mobile phase consisted of methanol-water-acetic acid(26: 23: 1). The detection wavelength was set at 334 nm. The column temperature was 25 degrees C. The flow rate was 1.0 mL x min . The linear response ranged within 2.5-50 microg for chlorogenic acid (r = 0.998), 2.5-25 microg for caffeic acid (r = 0.998) and 4.97-41.47 microg for linarin (r = 0.999), respectively. Recoveries were 100.8% with RSD 2.1% for chlorogenic acid, 96.2% with RSD 2.3% for caffeic acid and 103.7% with RSD 1.8% for linarin. There was a significant difference in the content of chlorogenic acid, caffeic acid, linarin among the samples. The content of chlorogenic in the sample from Fengdou Chongqing city was the highest in those from other places. The content of caffeic acid in the all samples is very low. The content of linarin in the samples from Jiangsu province and Anhui province almost reached the national standard in pharmacopoeia.

Determination of quercetin, luteolin, apigenin and acacetin inFlos Chrysanthemi Indici by RP-HPLC

To develop a RP-HPLC method for the determination of quercetin, luteolin, apigenin and acacetin in Flos Chrysanthemi indici. An Eclipse XDB-C18 column (4.6 mm x 250 mm, 5 microm) was used at 25 degrees C with the mobile phases of methanol-0.2% phosphatic acid in a gradient manner. The flow rate was set at 1.0 mL x min(-1). The detection wavelength was 350 nm. The linear response ranged from 1.02-20.48 mg x L(-1) for quercetin (r = 0.9994, n = 5), 1.03-20.54 mg x L(-1) for luteolin (r = 0.9992, n = 5), 1.12-22.40 mg x L(-1) for apigenin (r = 0.9995, n = 5), 1.01-20.22 mg x L(-1) for acacetin (r = 0.9998, n = 5), respectively. Recoveries were 101.3% with RSD 1.3% for quercetin, 100.62% with RSD 1.4% for luteolin, 98.42% with RSD 1.7% for apigenin and 99.02% with RSD 0.8% for acacetin. A significant difference (alpha = 0.01) among the contents of four flavonoids and total flavonoids was found. The method is quick, simple and repeatable for simultaneous determination of quercetin, luteolin, apigenin and acacetin in Flos Chrysanthemi Indici.

Determination of Camphor and Borneol in Flos Chrysanthemi Indici by UAE and GC-FID

In the work, ultrasonic-assisted extraction (UAE) followed by gas chromatography with flame ionization detector (GC-FID) is developed for the quantitative analysis of the bioactive components of camphor and borneol in a traditional Chinese medicine (TCM) of Flos Chrysanthemi Indici. The extraction parameters are investigated. The optimum extraction conditions found are: solvent, methanol; solvent to sample ratio, 12:1 (v/w); extraction time, 15 min. Camphor and borneol are determined using this extraction method in Flos Chrysanthemi Indici samples from 5 different growing areas. The relative standard deviation values for camphor and borneol are 8.4% and 5.6%, respectively. The recoveries for camphor and borneol are 89% and 95%, respectively, and the method detection limits are lower than 0.23 microg/mL. To demonstrate the method feasibility, steam distillation is also used to analyze camphor and borneol in Flos Chrysanthemi Indici samples from these different growing areas. The statistical comparison by t-test (95% confidence level) showed no significant difference between these results. It has been shown that the proposed UAE-GC-FID is a simple, rapid, and reliable method for quantitative analysis of camphor and borneol in TCM, and a potential tool for quality assessment of Flos Chrysanthemi Indici.

Determination of Camphor and Borneol in Traditional Chinese Medicines by Microwave-assisted Extraction and Gas Chromatography with Flame Ionization Detector

In this work, microwave-assisted extraction (MAE) followed by gas chromatography with flame ionization detector (GC–FID) was developed for the rapid determination of camphor and borneol in three traditional Chinese medicines (TCM): Chrysanthemi indici, Flos Chrysanthemi indici and Amomum villosum lour. The optimal MAE conditions obtained were: acetone for solvent, with solvent having sample ratio of 12:1 (v/w); microwave power of 380 W, and an irradiation time of 4 min. Method validations were also studied. To demonstrate the proposed method, ultrasonic-assisted extraction (UAE) and steam distillation (SD), followed by GC-FID, were used to analyze camphor and borneol in thethree TCMs. The close results were obtained by the three methods. The results showed that the proposed MAE–GC–FID is a simple, rapid, and reliable method for quantitative analysis of camphor and borneol in TCM, and is also a potential tool for TCM quality assessment.

Anticomplementary principles of a Chinese multiherb remedy for the treatment and prevention of SARS

Aim of the study To elucidate the anticomplementary principles of a Chinese multiherb remedy used for the treatment and prevention of SARS, which contains five ingredients of Herba Houttuyniae, Flos Chrysanthemi Indici, Herba Artemisiae Scopariae, Herba Eupatorii and Fructus Tsaoko. Materials and methods The anticomplementary activity was evaluated from hemolytic assays through the classical pathway (CP) and the alternative pathway (AP) of complement system in vitro. Compounds were isolated using bioactivity-guided fractionation and tested in vitro for their complement-inhibiting properties on the CP and AP. HPLC-DAD-ESI-MS was used to assign the isolated compounds in the five ingredient herbs. Results 15 compounds, including chlorogenic acid ( 1), rutin ( 2), hyperoside ( 3), p-hydroxyacephenone ( 4), scopoletin ( 5), quercitrin ( 6) (3 R,4 R,6 S)-3,6-dihydroxy-1-menthene ( 7), acaciin ( 8), scoparone ( 9), luteolin ( 10), quercetin ( 11), apigenin ( 12), acacetin ( 13), aristolactam ( 14), and apigenin-7,4′-dimethyl ether ( 15) were isolated and identified. Nine flavonoids ( 2, 3, 6, 8, 10– 13, 15) were found to show inhibitory effects towards the CP and AP of complement system, luteolin ( 10) was the most potent with the CH 50 and AP 50 values of 0.19 and 0.17 mM. The bioactive flavonoids were mainly derived from Herba Houttuyniae, Flos Chrysanthemi Indici and Herba Artemisiae Scopariae. Conclusions A Chinese multiherb remedy used for the treatment and prevention of SARS has robust inhibitory effect on complement system through the CP and AP, and its anticomplementary activity could be attributed to the flavonoids present in some ingredient herbs. Luteolin could be a potential anticomplementary agent.

A gradient HPLC method for the quality control of chlorogenic acid, linarin and luteolin in Flos Chrysanthemi Indici suppository

The Flos Chrysanthemi Indici suppository, a famous traditional Chinese medicine, is marketed for the treatment of prostatitis and chronic pelvic inflammation. A simple, relatively rapid and accurate high-performance liquid chromatographic (HPLC) method was applied to the simultaneous determination of chlorogenic acid, linarin and luteolin in Flos Chrysanthemi Indici suppository. The good separation was achieved on a Hypersil ODS column by a gradient elution using the mixture of acetonitrile and aqueous 1.0% acetic acid as mobile phase within 23 min. The method was validated for three active phenolic compounds with the relative standard deviations (R.S.D.) of intra- and inter-day precision below 3.0%, and the detection limits (S/N = 3) of chlorogenic acid, linarin and luteolin were 0.32, 0.08 and 0.05 μg ml −1, respectively. This assay was successfully applied to the determination of three active phenolic compounds in Flos Chrysanthemi Indici suppository samples with the quantitative recoveries in the range of 94.7–101.8%. The results indicate that the developed HPLC method can be readily utilized as a quality control method for Flos Chrysanthemi Indici suppository and its related traditional Chinese medicinal preparations.

Development of microwave-assisted extraction followed by headspace solid-phase microextraction and gas chromatography–mass spectrometry for quantification of camphor and borneol in Flos Chrysanthemi Indici

In the work, microwave-assisted extraction (MAE) followed by headspace solid-phase microextraction (HS-SPME) and gas chromatography–mass spectrometry (GC–MS) was developed for quantitative analysis of the bioactive components of camphor and borneol in a traditional Chinese medicines (TCM) of Flos Chrysanthemi Indici. After systematical investigation, the optimal experimental parameters microwave power (400 W), irradiation time (4 min), fiber coating (PDMS/DVB fiber), extraction temperature (40 °C), extraction time (20 min), stirring rate (1100 rpm), and salt effect (no salt added) were investigated. The optimized method provided satisfactory precision (RSD values less than 12%), good recovery (from 86% to 94%), and good linearity ( R 2 > 0.999). The proposed method was applied to quantitative analysis of camphor and borneol in Flos Chrysanthemi Indici samples from 11 different growing areas. To demonstrate the method feasibility, steam distillation was also used to analyze camphor and borneol in Flos Chrysanthemi Indici samples from these different growing areas. The very close results were obtained by the two methods. It has been shown that the proposed ME–HS-SPME–GC–MS is a simple, rapid, solvent-free and reliable method for quantitative analysis of camphor and borneol in TCM, and a potential tool for quality assessment of Flos Chrysanthemi Indici.

Quality assessment of Flos Chrysanthemi Indici from different growing areas in China by solid-phase microextraction-gas chromatography-mass spectrometry

Flos Chrysanthemi Indici is a common traditional Chinese medicine (TCM). In this paper, headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) was developed for quality assessment of Flos Chrysanthemi Indici from different growing areas in China. SPME parameters such as extraction fibers, extraction temperature, extraction time and sample mass were investigated to achieve identical results to those obtained by the steam distillation (SD). The selected SPME conditions were as follows: SPME fiber coated with 65-μm PDMS/DVB, extraction temperature of 60 °C, extraction time of 30 min and sample mass of 1.0 g. Furthermore, four active compounds (eucalyptol, camphor, borneol and bornyl acetate) presented in the TCM were applied to evaluating the quality of Flos Chrysanthemi Indici from 20 various areas. The quality assessment was successfully performed to compare the similarity value ( S ) between different sample vector of Flos Chrysanthemi Indici and the standard profile vector (SPV). The results showed that the proposed HS-SPME-GC-MS was an alternative technique for quality assessment of Flos Chrysanthemi Indici samples.

Quality assessment of Flos Chrysanthemi Indici from different growing areas in China by solid-phase microextraction-gas chromatography-mass spectrometry

Flos Chrysanthemi Indici is a common traditional Chinese medicine (TCM). In this paper, headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) was developed for quality assessment of Flos Chrysanthemi Indici from different growing areas in China. SPME parameters such as extraction fibers, extraction temperature, extraction time and sample mass were investigated to achieve identical results to those obtained by the steam distillation (SD). The selected SPME conditions were as follows: SPME fiber coated with 65-μm PDMS/DVB, extraction temperature of 60 °C, extraction time of 30 min and sample mass of 1.0 g. Furthermore, four active compounds (eucalyptol, camphor, borneol and bornyl acetate) presented in the TCM were applied to evaluating the quality of Flos Chrysanthemi Indici from 20 various areas. The quality assessment was successfully performed to compare the similarity value ( S) between different sample vector of Flos Chrysanthemi Indici and the standard profile vector (SPV). The results showed that the proposed HS-SPME-GC-MS was an alternative technique for quality assessment of Flos Chrysanthemi Indici samples.

Solid-Phase Microextraction Followed by Gas Chromatography-Mass Spectrometry Analysis of the Volatile Components of Flos Chrysanthemi indici in Different Growing Areas

Headspace solid phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) has been applied to the determination of the volatile compounds of a common traditional Chinese medicine , Flos Chrysanthemi indici. Twenty-six compounds were identified by the present method, which are similar to the results from steam distillation. Difference in analysis from ten growing areas was found to be in their relative content and total concentrations. Four active compounds (cineole, 3-thujanone, borneol and bornyl acetate) were applied to the evaluation of the quality of Flos Chrysanthemi indici from different areas. By comparing their peak areas, it was found that Flos Chrysanthemi indici with the best quality was from Hangzhou in China. These results show that HS-SPME-GC-MS is a simple, rapid and solvent-free method for the analysis of the volatile compounds in this material and a potential tool for its quality assurance.