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Neurodegenerative disorders (NDD) have become the common global health burden over the last several decades. According to World Health Organization (WHO), a staggering 30 million people will be affected by Alzheimer's disease in Europe and the USA by 2050. Effective therapies in this complex field considering the multitude of symptoms associated with NDD indications, have not been found yet. Based on the results of NDD related studies, prevention appears to be the promise alternative. Antioxidative and anti-inflammatory properties are hypothesized for natural phenolics, a group of plant secondary products that may positively impact neurodegenerative diseases. In these studies, phenolic-rich extracts from less common fruit species: Blue honeysuckle (Lonicera edulis, Turcz. ex. Freyn), Saskatoon berry (Amelanchier alnifolia Nutt.), and Chinese hawthorn (Crateagus pinnatifida Bunge) were obtained and analyzed to detect neuroprotective substances content and establish a potential therapeutic value. High performance liquid chromatography with electrochemical detection was optimized and further applied on analysis of the extracts of less common fruit species. It was observed that Chinese hawthorn and Blue honeysuckle extracts are potent source of neuroprotective phenolic antioxidants. In accordance the results, it appears that the fruit or formulated products may have the potential for the prevention of neurodegenerative diseases.

Photodynamic therapy (PDT) is an effective therapy for local malignant tumors. Lonicera japonica was found to have the anti-tumor effect. The aim of this study is to explore the mechanisms of apoptosis induced by PDT in lung CH27 carcinoma cells with alcohol extract from Lonicera japonica as photosensitizer. Our study indicated that Lonicera japonica extracts exhibited significant photocytotoxicity in CH27 cells at a concentration range of 50-150 microg/ml, with 0.4-1.2J/cm2 light dose. PDT with Lonicera japonica extracts-induced cell death is a typical apoptosis that was accompanied by DNA condensation, externalization of phosphatidylserine and formation of apoptotic bodies. PDT with Lonicera japonica extracts was shown to be caspase-3-independent apoptosis via activation of AIF in this study. P38-associated pathway may be involved in apoptosis induced by PDT with Lonicera japonica extracts in CH27 cells. We also have demonstrated that PDT with Lonicera japonica extracts-induced CH27 cells apoptosis was probably related to its ability to change the protein expression and distribution of heat shock protein 27.

A rapid and simple method has been developed for the screening and identification of natural antioxidants of Flos Lonicerae Japonicae (FLJ), derived from the flower buds of Lonicera japonica. The hypothesis is that upon reaction with 1,1-diphenyl-2-picrylhydrazyl (DPPH), the peak areas (PAs) of compounds with potential antioxidant effects in the HPLC chromatograms will be significantly reduced or disappeared, and the identity confirmation could be achieved by HPLC-DAD-TOF/MS hyphenated technique. Using the proposed approach, about 14 compounds in the FLJ extract were found to possess a potential antioxidant activity. They were identified as chlorogenic acid (1), 1-O-caffeoylquinic acid (1-O-CQA, 2), caffeic acid (4), 4-O-CQA (5), rutin (7), isoquercitrin (8), luteolin-7-O-glucoside (9), lonicerin (10), 4,5-O-dicaffeoylquinic acid (4,5-O-diCQA, 11), 3,5-O-diCQA (12), 1,3-O-diCQA (13), 3,4-O-diCQA (14), 1,4-O-diCQA (16), and luteolin (17). In addition, the free radical scavenging capacities of the available identified compounds were also investigated by HPLC assay. The results indicated that the compounds with PAs significantly decreasing were natural antioxidants, whereas those with PAs not changing presented no activities, which accordingly indicated that this newly proposed method could be widely applied for rapid screening and identification of natural antioxidants from complex matrices including Chinese herbal medicines.

A method which involves combination of centrifugal ultrafiltration sampling with high-performance liquid chromatography coupled with diode array detection and mass spectrometry (HPLC-DAD-MS) analysis was established for screening bioactive compounds binding to calf thymus deoxyribonucleic acid (ct-DNA) from the extracts of Lonicera japonica. Four compounds were screened out and identified as rutin, quercetin-3-O-glucoside, luteolin-7-O-glucoside and lonicerin, based on the comparison of retention time, UV spectra and MS data with those of standards. The DNA-binding capabilities of the latter three flavonoids were found for the first time. The binding mechanisms of rutin, quercetin-3-O-glucoside and luteolin-7-O-glucoside with ct-DNA at the molecular level were explored using acridine orange (AO) as a fluorescence probe. Groove binding is the most appropriate binding mode of these three flavonoids to DNA, according to ultraviolet absorption and fluorescence spectra, as well as melting temperature (T(m)) curves and viscosity measurements. The binding constants of rutin, quercetin-3-O-glucoside and luteolin-7-O-glucoside with DNA-AO complex were 3.81 x 10(3), 3.37 x 10(3) and 5.50 x 10(3) L/mol, respectively.

Objective: To study and establish the GC fingerprint of essential oils of 10 batches the flower of Lonicera japonica., Method: The essential oils were extracted by steam distillation from ten batches of the flowers, then separated by capillary gas chromatography., Result: The similarity of 10 batches of the flower of L. japonica was high., Conclusion: The method was reliable, and can be used to evaluate the quality of the flower of L. japonica.

Two new triterpenoid saponins, loniceroside D (1) and loniceroside E (2), were isolated from the dry flowers and buds of Lonicera japonica Thunb., along with seven known compounds, chlorogenic acid (3), sweroside (4), vogeloside (5), epi-vogeloside (6), loniceroside A (7), loniceroside B (8), and loniceroside C (9). Their structures were elucidated on the basis of spectroscopic data, physicochemical properties, and acid hydrolysis.

A novel cyclic peroxide, shuangkangsu (1), has been obtained from the water extract of the buds of Lonicera japonica (Thunb.). The structure was elucidated as 5,8-divinyl-1,4-dihydro-1,4-di-O-beta-d-glucopyranosyl-2,3-dioxane (1) on the basis of the spectral data. Its absolute stereochemistry was determined to be (1S, 4S) by comparison of its CD curves with those of its optically pure analogs 2 and 3, which were synthesized from the phthalaldehyde. The absolute configurations of 2 and 3 were determined to be (1R, 4R) and (1S, 4S) by X-ray analysis and CD spectra, respectively. Compound 1 showed significant antiviral activities against influenza virus in chicken embryo and respiratory syncytial virus in cells.

Object: To establish the fingerprint spectrum of Lonicera macranthoides by HPLC., Methods: The column of Hypersil gold C18 (4. 6 mm x 250 mm, 5 microm) was used. The mobile phase consisted of Acetonitril-0.05% phosphoric acid with gradient elution. The column temperature was 30 degrees C, the detective wavelength was 240 nm, and the flow rate was 0.5 ml/min., Results: Fingerprint spectrum of Lonicera macranthoides was established, and 11 samples of different origin Lonicera macranthoides were detected. Ten peaks in the chromatogram were common by Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine (Version 2004 A). There was a high similarity and each chromatographic peak was obtained with good separation and correlation according to the technical requirements of fingerprint of Chinese traditional medicine., Conclusion: This method is accurate, reliable and provides a scientific basis for controlling the quality of Lonicera macranthoides.

Objective: To study the chemical constituents of flower buds of Lonicera macranthoides., Methods: The 90% EtOH extract of Lonicera macranthoides. was successively partitioned with petroleum ether and ethyl acetete., Results: Repeated column chromatography of the ethyl acetete fraction afforded the following compounds (1-9): ginnol (1), triacontanol (2), ursolic acid (3), beta-sitosterol (4), triacontane (5), palmitic acid (6), beta-daucosterol (7), 3-decyl-3-octyldocosan-1-ol (8), 3-dodecyl-3-nonyldocosan-1-ol (9). All compounds except 4 are isolated from this plant for the first time while compounds 2, 3, 5, 8 and 9 are their first time been isolated from genus Lonicera.

This study aimed to elucidate some novel pharmacological activities of Lonicera japonica (Caprifoliaceae), which is widely used in Oriental folk medicine. The ethanolic extract of L. japonica (LJ) dose dependently inhibited chick chorioallantoic membrane angiogenesis. The antinociceptive activity of LJ was assessed using the acetic acid-induced constriction model in mice. LJ showed anti-inflammatory activity in two in-vivo models: the vascular permeability and air pouch models. LJ suppressed the production of nitric oxide via down-regulation of inducible nitric oxide synthase in lipopolysaccharide-stimulated RAW264.7 macrophage cells. However, LJ was unable to suppress induction of cyclooxygenase-2 in the stimulated macrophage cells. LJ decreased the reactive oxygen species level in the stimulated macrophage cells. In brief, the flowers of L. japonica possess potent anti-angiogenic and antinociceptive activities, in addition to anti-inflammatory activity, which partly supports its therapeutic efficacy.

Chlorogenic acid, one of the most bioactive compounds rich in the Chinese medicinal herb honeysuckle, is a natural antioxidant and serves as anti-inflammatory, anti-tumor, anti-mutagenic and anti-carcinogenic agent. An efficient preparative separation process of chlorogenic acid from honeysuckle crude extracts has been developed in the present study. HPD-850 resin offers the best adsorption capacity, and adsorption and desorption ratios for chlorogenic acid among the nine macroporous resins tested, and its adsorption rate at 25 degrees C fit best to the Langmuir isotherm. The adsorption capacity of HPD-850 resin was found to depend strongly on the pH value of the initial adsorption solution. The dynamic adsorption and desorption experiments have been carried out on a HPD-850 resin packed column to optimize the separation process of chlorogenic acid from honeysuckle crude extracts. After one run treatment with HPD-850 resin, the chlorogenic acid content in the final product was increased 4.46-fold from 11.2% to 50.0%, with a recovery yield of 87.9%. The preparative separation of chlorogenic acid can be easily and efficiently achieved via adsorption and desorption on HPD-850 resin, and the method developed will provide a potential approach for large-scale separation and purification of chlorogenic acid for its wide pharmaceutical use.

Three pyridinium inner salt alkaloids, lonijaposides A-C (1-3), based on an unprecedented skeleton of an N-substituted nicotinic acid nucleus coupled through C-5 with C-7 of a secoiridoid, together with seven known iridoids, have been isolated from the flower buds of Lonicera japonica. Their structures were elucidated by spectroscopic and chemical analyses. Lonijaposide C (3) showed activity against the release of glucuronidase in rat polymorphonuclear leukocytes induced by the platelet-activating factor.

Objective: To establish a chromatographic fingerprint of Lonicera japonica and apply it to identify geo-authentic of L. japonica., Method: The HPLC was applied in chromatographic separation and data were analysed by "computed aided similarity evaluation" software., Result: There exited distinct difference of chemical components group between and non-authentic samples., Conclusion: The established HPLC fingerprint can be used for the identification of geo-authentic of L. japonica.

Objective: To search for the optimal process of Flos Lonicerae in producing area, in order to offer scientific and applied process method for steadying the quality of Flos Lonicerae., Methods: To summarize and use for reference the reported process method, searching for the relativity of different processes and quality., Results: The content of Chlorogenic Acid in kill-enzyme torrefaction sample was 12. 8% higher than directly dried in the sun and 24.9% higher than dried in the shade. The content of Luteolin-7-glu in kill-enzyme torrefaction sample was 7.8% higher than the directly dried in the sun and 54.3% higher than dried in the shade., Conclusion: At present, kill-enzyme torrefaction is the optimal process of Flos Lonicerae in producing area It is an important technology in GAP large-scale of Flos Lonicerae.

Objective: To determine the contents of chlorogenic acid and total caffeoylquinic acid in 6 Folium Lonicerae in Sichuan province and Chongqing city for evaluating the quality of them., Methods: The HPLC analysis and spectrophotometry analysis were used., Results: The contents of chlorogenic acid and total caffeoylquinic acid in 6 Folium Lonicerae had been determined., Conclusion: The method is accurate, simple and has a good reproducibility. The results can be used as a reference for the reasonable medication, quality control and further study of Folium Lonicerae.

Five caffeoylquinic acid derivatives (CQAs), including methyl 3,4-di-O-caffeoylquinate (3,4-diCQM), methyl 3,5-di-O-caffeoylquinate (3,5-diCQM), 3,4-di-O-caffeoylquinic acid (3,4-diCQA), 3,5-di-O-caffeoylquinic acid (3,5-diCQA) and chlorogenic acid (CA), were isolated from Lonicera fulvotomentosa HSU et S. C. CHENG to be used as model compounds. The binding of these bioactive components to bovine serum albumin (BSA) was investigated by fluorescence quenching method. The results showed that there were binding affinities for CQAs with BSA, and the binding constants ranked in the following order: 3,4-diCQM>3,5-diCQM<3,4-diCQA>3,5-diCQA>CA, under the physiological conditions, which suggested that the numbers and the substituted positions of caffeoyl group as well as the esterification of carboxyl group in the molecular structures appeared to contribute moderate effects to the interaction processes. Furthermore, the Stern-Volmer curves demonstrated that CQAs caused the fluorescence quenching through a static quenching procedure. Thermodynamic analysis indicated that both hydrophobic and electrostatic interactions played major roles in stabilizing the complex. The binding distance for each binding reaction was also calculated by the Föster theory.

A new pH-gradient counter-current chromatography method for the isolation of chlorogenic acid from flowers and buds of Lonicera japonica Thumb. has been successfully established using a novel upright coil planet centrifuge with three multi-layer coils connected in series with 600 mL capacity. The crude extracts were first prepared by direct extraction with hot water and following concentration to remove the solution. Then the two-phase solvent system composed of ethyl acetate-n-butanol-water (2:1:3, v/v) was applied to the separation. Its neutral upper phase was used as stationary phase, whereas both its neutral lower phase and base lower phase with 10mM NH(3) were employed as mobile phase with gradient elution in the head to tail mode. As a result, 330 mg quantity of crude extract was purified in one-step separation for 180 min, yielding 20.5mg chlorogenic acid with over 98% purity. Structure of the compound is further identified by electrospray ionization tandem mass spectrometry (ESI-MS/MS) and nuclear magnetic resonance (NMR).

To investigate the chlorogenic acids resources in Eucommia ulmoides Olive leaves, Lonicera japonica Thunb. leaves and Houttuynla cordata Thunb. leaves, methanolic extracts of these three materials have been analysed qualitatively for chlorogenic acids and their derivatives by structure-diagnostic LC-MSn. Three monocaffeoylquinic acids (3-CQA, 4-CQA, 5-CQA) were detected. 5-CQA dominated this subgroup in Eucommia ulmoides Olive and Lonicera japonica Thunb. leaves, but 3-CQA and 4-CQA dominated this subgroup of chlorogenic acids in the leaves of Houttuynla cordata Thunb. Caffeoylquinic acid-glycosides were detected for the first time from Eucommia ulmoides leaves. 5-FQA was found in Lonicera japonica Thunb. leaves, and 3-FQA and 4-pCoQA have been identified in Houttuynla cordata Thunb. This is the first report of the chlorogenic acid profile in Houttuynla cordata Thunb. The comparatively unusual profile of caffeoylquinic acids in Houttuynla cordata Thunb. makes it a convenient source of 3-CQA and 4-CQA that are not commercially available.

The screening and analysis of bioactive components in traditional Chinese medicines (TCMs) is very important not only for the quality control of Chinese herbs but also for elucidating the therapeutic principles. This study developed a new method for screening and analyzing bioactive compounds from TCMs using centrifugal ultrafiltration coupled with high-performance liquid chromatography. The method was successfully applied in the binding study of Flos Lonicerae Japonicae with bovine serum albumin (BSA), and 11 compounds were found to be bound with the BSA. Eight of them were positively identified as chlorogenic acid, caffeic acid, rutin, quercetin-3-O-glucoside, luteolin-7-O-glucoside, lonicerin, 3, 5-di-O-caffeoyl quinic acid and 3,4-di-O-caffeoyl quinic acid. Another three compounds were tentatively identified as two isomers of chlorogenic acid and one isomer of di-O-caffeoyl quinic acid by comparing the UV data and MS data with the previous reports. Based on modern pharmacological study, these compounds are the major bioactive components in Lonicera japonica. Therefore, the proposed method could be a good approach to predicting the potential bioactivities of multiple compounds in TCMs simultaneously., ((c) 2007 John Wiley & Sons, Ltd.)

Lonicera maackii is an invasive shrub in North America for which allelopathic effects toward other plants or herbivores have been suspected. We characterized the major phenolic metabolites present in methanol extracts of L. maackii leaves. In addition, we examined the effects of methanol-water extracts of L. maackii leaves on seed germination of a target plant species and on feeding preference and growth rate of a generalist insect herbivore. A total of 13 individual major and minor compounds were detected in crude leaf extracts by high-performance liquid chromatography coupled to electronspray ionization-tandem mass spectrometry (ESI-MS/MS). Extracts were dominated by two major flavones, apigenin and luteolin, and their glucoside derivatives, apigenin-7-glucoside and luteolin-7-glucoside. Quantities of these compounds, along with chlorogenic acid, varied between two sampling points. Leaf extracts that contained these compounds were inhibitory to seed germination of Arabidopsis thaliana. In addition, treatment of artificial diet with leaf extracts deterred feeding of the generalist herbivore, Spodoptera exigua, in choice experiments but had no effect on growth rate in short-term no-choice bioassays. Purified apigenin tended to deter feeding by S. exigua and inhibited seed germination of A. thaliana. We conclude that leaves of L. maackii contain phenolic compounds, including apigenin and chlorogenic acid, capable of having biological effects on other plants and insects.

Interaction of the Flos Lonicerae Japonicae (FLJ) with protein was studied by microdialysis coupled with HPLC-DAD-MS. Eight compounds were identified by comparing their t(R), UV data and MS data with reference compounds. Microdialysis recoveries and binding degrees of compounds in FLJ with bovine serum albumin (BSA) were determined. Recoveries of microdialysis sampling ranged from 51.3 to 73.2% with relative standard deviation (RSD) below 3.1%, and the binding degrees of those compounds to BSA ranged from 4.8 to 61.2 (0.3 mM BSA) and from 11.1 to 76.2% (0.6 mM BSA), respectively. The results showed that the binding properties of compounds in FLJ were influenced by pH. Furthermore, the binding degrees of five reference compounds were determined separately under the same conditions, the binding degrees of chlorogenic acid, luteolin-3-O-glucoside and 4,5-di-O-caffeoyl quinic acid was lower in FLJ than in single compound solution, on the contrary the binding degree of caffeic acid and rutin was higher in FLJ, which indicated that a significant effects of the interaction of compounds with each other on their binding degrees to BSA. The results showed that there were ten compounds had interaction with BSA, eight of them were the proven active compounds, and the other two compounds had similar binding degree with the proven active compounds, so the ten compounds might possess potential activities.

Background: Lonicera caerulea L. (blueberry honeysuckle, Caprifoliaceae) is a traditional crop in northern Russia, China, and Japan. Its fruits are little known as edible berries in North America and Europe. This review deals with the botany and chemical composition of L. caerulea and the biological activity of its main constituents, focusing on the potential health benefits of the berries., Methods and Results: PubMed, Science Direct and ISI Web of Knowledge databases were used for this paper. Literature sources include the period 1935-2007. L. caerulea berries a are rich source of phenolic compounds such as phenolic acids as well as anthocyanins, proanthocyanidins and other flavonoids, which display potential health promoting effects. Chemopreventive, antimicrobial, anti-adherence and antioxidant benefits, among others are described for these compounds., Conclusions: The potential of L. caerulea berries to prevent chronic diseases such as diabetes mellitus, cardiovascular diseases and cancer seems to be related above all to their phenolic content.

Objective: To study the different manufacturers Vitmin C (Vc) Yinqiao tablets, and the quality of the analysis of the problem, to provide a theoretical basis for the correct evaluation of the quality of medicines and improving the standard drugs., Method: 11 manufacturers of 18 batches of samples for determination of the weight of the core tablets, powder samples were observed with microscope, determination of Vc, and the establishment of the Vc Yinqiao tablets HPLC method for determination of chlorogenic acid and arctigenin, chlorogenic acid and arctigenin in the samples were measured and compared., Result: There is a big difference of microscope and various measured results in different manufacturers products., Conclusion: Because different manufacturers to produce the same, there are big differences in the quality of the products.

Objective: Utilizing RP-HPLC analysis of single, merging and simultaneously boiled Lonicera japonica and Forsythia suspense to find out their identity and difference., Method: Samples were separated by Alltech-C18 column (4. 6 mm x 250 mm, 5 microm) with a mixture solution of acetonitrile-water (containing 0.2% ethanoic acid) as mobile phase, 1.0 mL x min(-1) as flow-rate, 280 nm as detected wave-length, room temperature as temperature of column, 10 microL as injected volume., Result: The linear ranges of chlorogenic acid and forsythoside A were 0.980-9.800 microg (r = 0.999 9) and 0.972-9. 720 microg (r = 0.999 9), respectively. The average recoveries of chlorogenic acid and forsythoside A were 99.85% (n = 6, RSD 2.0%) and 100.87% (n = 6, RSD 1.6%), respectively. Major peaks were confirmed to the two component herbs, contents of chlorogenic acid and forsythoside A were determinated, ingredients were represented by following peaks were compared qualitatively: peaks k, m, n: L. japonica > merging sample > boiled sample; peaks g, h: Forsythia suspense > merging sample > boiled sample; peaks d, e, i: boiled sample > L. japonica, boiled sample > merging sample., Conclusion: Difference and identities among 4 experimental samples of RP-HPLC separations show that Yinqiao, L. japonica and F. suspense' substances are difference and identities, which offered direction for investigating further the topic-pharmacodynamic substance of Yinqiao, provided increasing experimental evidence for clinic medicine and pharmaceutics and supplied reference for the complex prescription of traditional Chinese medicine.

Objective: To develop an HPLC method for determination of loniceroside A and loniceroside C in Lonicerae japonicae., Method: The analysis was carried out on an Alltech C18 column (4.6 mm x 250 mm, 5 microm) evaluated with methanol-acetonitrile-0.1% glacial acetic acid as mobile phase. Flow rate was at 1.0 mL x min(-1) and the detection wavelength was at 210 nm for UV detection., Result: The calibration curves were linear over the range from 0.15 to 2.25 microg (r = 0.999 9) for loniceroside A and 0.11 to 1.65 microg (r = 0.999 1) for loniceroside C. The average recoveries were 99.9% and 98.3%, respectively., Conclusion: The contents of Loniceroside A&C are diverse in Flos Lonicerae japonicae in different regions.

To develop and validate an ultra performance liquid chromatography tandem mass spectrometric (UPLC-MS/MS) method for the simultaneous quantification of baicalin and chlorogenic acid in human plasma after iv infusion of Yinhuang injection, the analytes were isolated from plasma by protein precipitation with methanol. Then they were chromatographied on an Acquity UPLC BEH C18 column (50 mm x 2.1 mm ID, 1.7 microm) at 40 degrees C. The mobile phase A consisted of water and 0.1% formic acid. The mobile phase B consisted of methanol and 0.1% formic acid. The analytes were eluted from the column with a linear gradient from 5% B to 80% B in 5 min, then hold for 0.5 min before returning to initial condition. The flow rate was 0.35 mL x min(-1). A tandem mass spectrometer equipped with electrospray ionization source was used as detector. Multiple reaction monitoring (MRM) using the precursor to product ion pairs of m/z 447-->271 (for baicalin), m/z 353-->191 (for chlorogenic acid) and m/z 287-->287 (for internal standard) were used to quantification. The linear concentration ranges of the calibration curves for baicalin and chlorogenic acid ranged from 9.6 to 1540 ng x mL(-1) and from 7.5 to 1200 ng x mL(-1), respectively. The intra- and inter-day relative standard deviation (RSD) across three validations run over the entire concentration range was less than 10.2% for both baicalin and chlorogenic acid. After iv infusion of Yinhuang injection to the volunteers, the concentration-time curves of baicalin and chlorogenic acid fitted the two-compartment and three-compartment model. T(1/2)alpha were (4.47 +/- 0.89) and (7.65 +/- 4.42) min, T(1/2)beta were (46.22 +/- 10.03) and (34.40 +/- 19.16) min, respectively. The method was proved to be highly sensitive, selective, and suitable for pharmacokinetic investigations of both baicalin and chlorogenic acid.

Objective: To compare HPLC fingerprint of Flos Lonicerae Japonicae in different collecting time for determining harvest., Methods: The data from RP-HPLC were analyzed by similarity evaluation, principle component analysis and system cluster analysis., Results: The content of chemical composition of Flos Lonicerae Japonicae were vaiance on collecting time. The best harvesting time was the erbai period., Conclusion: The study offers the basis of controlling the quality of Flos Lonicerae Japonicae and the realization of good agricultural practice.

Objective: To observe the protective effect of Jinhuang-1 (JH-1) on oleic acid-induced acute respiratory distress syndrome (ARDS) in rats and provide a basis to clinical application., Method: Oleic acid was injected into the tail vein of rats and JH-1 were administered to rats. After four days, their blood were collected for the blood gas analysis. The superoxide dismutase (SOD), the malondialdehyde (MDA), nitic oxide (NO) content and the positive expression ratio of tumor necrosis factor-alpha (TNF-alpha) were determined. The pathological changes of microstructure of lung were observed., Result: The results demonstrated that JH-1 could significantly increase PO2, reduce PCO2 of oleic acid-induced ARDS rats, increase the superoxide dismutase, decrease the malondialdehyde, nitic oxide content and the positive expression ratio of TNF-alpha, and improve the histological destruction on lung., Conclusion: JH-1 plays a protective role for ARDS rats induced by oleic acid. The mechanism is probably related to attenuating lipid peroxidation and decreasing NO content and the expression of TNF-alpha in lung.

Antioxidants are emerging as prophylactic and therapeutic agents for various diseases. However, little is known about the antioxidant property of the extract from Flos Lonicerae, a medically useful traditional Chinese medicine herb. Here the antioxidant capacity of water, methanolic and 70% ethanolic extracts prepared from Flos Lonicerae to scavenge DPPH radical and reduce Fe(3+) to Fe(2+) is evaluated. Chlorogenic acid, a major component of Flos Lonicerae, is identified and further purified from 70% ethanolic extract with HPLC and its antioxidant capacity is also characterized. The content of total phenolic compounds and chlorogenic acid in Flos Lonicerae is determined. The present results demonstrate that all Flos Lonicerae extracts examined here exhibit antioxidant activity and chlorogenic acid is a major contributor to this activity, which implicates that the Flos Lonicerae extract may serve as potential source of natural antioxidants for treatment of some diseases.

Chlorogenic acid (ChA), a major phenolic compound in the Flos Lonicerae, is widely used in the traditional Chinese medicine practice. The purpose of this study is to report the pharmacokinetic parameters of ChA in rats after oral administration and explore its absorption profile briefly. A two-compartment model was proposed and validated through the program to explain the apparent triphasic phenomenon of ChA in rats after intragastric administration. A rapid absorption and a relatively slow distribution followed by a slower elimination phase were observed. At the administered doses of 200, 400 and 600 mg/kg, the values of absorption half-life (t 1/2 Ka) were 10.23, 18.66 and 28.13 min. The values of distribution half-life (t1/2) were 12.35, 31.04 and 39.19 min. And the values of elimination half-life (t1/2P) were 231.64, 337.23 and 420.81 min. The volume of distribution at the three doses were 55.26, 35.56, 32.22 L/kg, respectively. The AUC0-->infinity (area under the concentration-time curve) was not proportional to the administered dose. In the range of the doses examined, the absorption pharmacokinetics of ChA in rats was based on nonlinear kinetics.

Objective: To study the influence of steaming processing on components in Chinese medicine., Methods: RP-HPLC was used to separate and detect every, components and all the same components' peak area were compared between crude-sun-cured and stcam-sun-cured Chinese medicine., Results: Steaming processing showed some advantatages to reserve most but not every compo-nent., Conclusion: Steaming processing can be applied as a superior method.

This study presents a new HPLC method for the simultaneous determination of seven major components, namely chlorogenic acid, caffeic acid, loganin, sweroside, secoxyloganin, rutin and luteolin 7-O-glucoside in Caulis Lonicerae Japonicae, a commonly used traditional Chinese medicinal herb derived from the caulis of Lonicera japonica Thunb. These seven compounds, belonging to the chemical types of phenolic acids, iridoids and flavonoids, were separated on a C18 column (250 x 4.6 mm, 5.0 microm) with the column temperature at 30 degrees C. The mobile phase was composed of (A) aqueous acetic acid (0.4%, v/v) and (B) acetonitrile using a gradient elution of 10% B at 0-12 min, 10-17% B at 12-25 min and 17% B at 25-35 min. The flow rate was 1.0 mL/min and detection wavelength was set at 245 nm. The limit of detection (S/N = 3) ranged from 0.10 to 0.23 microg/mL and the limit of quantification (S/N = 10) ranged from 0.69 to 3.56 microg/mL. All calibration curves showed good linear regression (r2 > 0.9990) within the test ranges. The intra- and inter-day precisions as determined from sample solutions were below 1.24 and 2.28%, respectively. The recoveries for seven compounds were found to range from 94.2 to 103.6%. This verified method has been successfully applied to evaluation of commercial samples of Caulis Lonicerae Japonicae from different markets in China., (Copyright 2007 John Wiley & Sons, Ltd.)

Honeysuckle is a lonicera plant, commonly used traditional precious raw material for medicine in China, and is a commonly used Chinese native medicine and an important material for cold drink. Honeysuckle has the effect of alleviating fever and disintoxicating, and is used mainly to treat the carbuncular swelling, and cure boils, throat rheumatism, erysipelas, painfully hot sun blood dysentery, cold, and various diseases accompanied by fever. The simultaneous determination of multi-metal elements in honeysuckle was studied by inductively coupled plasma atomic emission spectrum (ICP-AES) after carbon pick-up acid dissolvation digestion. The average recovery of the method for Cu, Mn, Ni, Pb, Fe, Cr, Ca and Mg in honeysuckle was found to be 83.8%-115.6%, while RSD (n = 10) is 1.1%-4.9% for the multi-elements in honeysuckle. The method was applied to the analysis of some practical samples and the results obtained were satisfactory. Fe and Mn are rather rich in honeysuckle. It is of important significance to assess the value of the food cooked with medicinal herbs by the determination of multi-trace elements.

The anti-inflammatory properties of aqueous extracts from Lonicera japonica (LJ) flower, an anti-inflammatory treatment in traditional Chinese medicine, were tested by radioimmunoassay of cyclooxygenase isoenzyme-generated prostaglandin E2 (PGE2) synthesis as well as by Western and Northern blot analysis of COX-2 protein and mRNA expression, respectively. Boiled LJ aqueous extracts directly inhibited both COX-1 and COX-2 activity, while non-boiled extracts stimulated COX-1. Boiled LJ extracts also inhibited expression of IL-1beta-induced COX-2 protein expression and suppressed its mRNA induction by IL-1beta in A549 cells. Suppression of COX-2 mRNA induction required a significantly higher dose of aqueous extract than did suppression of protein expression, indicating that compounds in the extract act translationally or post-translationally at lower doses and transcriptionally or post-transcriptionally at higher doses. Direct inhibition of COX isoenzymes as well as down-regulation of COX-2 mRNA and protein may represent the mechanism by which this ancient herbal treatment decreases inflammation.

Objective: To compare the quality of Flos Lonicerae between different producing areas., Method: ICP-AES, UV and HPLC were used to determine the contents of trace elements, chlorogenic acid, total flavonoids, five iridoid glucosides, hederagenin, and oleanolic acid. SAS software system was used to perform data and cluster analyses., Result: The results showed that the geo-authentic crude drug was lower in the contents of Cr and Pb but higher in the contents of chlorogenic acid, total flavonoids, five iridoid glucosides, hederagenin, and oleanolic acid than the non-authentic crude drug., Conclusion: The geo-authentic crude drug of Flos Lonicerae is better in quality than the non-authentic crude drug based on the modern chemical analyses, which confirms the validity of traditional geo-based classification.

Objective: To establish a rapid method for the determination of chlorogenic acid in aqueous solution of Flos Lonicerae Japonicae extraction., Method: Chlorogenic acid content in aqueous solution of Flos Lonicerae Japonicae extraction was measured by RP-HPLC. The UV spectrum of the same samples were scanned simultaneously. The wavelength of the best correlation between UV absorbance and chlorogenic acid content determined by HPLC was chosen and the chlorogenic acid content of aqueous solution of Flos Lonicerae Japonicae extraction was estimated by prediction equation. Other 10 batches of aqueous solution of Flos Lonicerae Japonicae extraction was used for testing the equation by comparison of the values of chlorogenic acid content determined by HPLC and equation., Result: Good linearity of chlorogenic acid was obtained within the range of 0.09-2.25 microg by HPLC; The recovery was 100.4%; RSD was 1.1% (n = 5). The best wavelength in UV spectrum correlating with chlorogenic acid content was 294 nm. The prediction equation: chlorogenic acid content C = 2.703 425A(294 nm) - 0.129 18; Good correlation between chlorogenic acid content determined by HPLC and predicted results was found (r = 0.995 2, n = 11). RSD was 0.2% (n = 6)., Conclusion: This method is simple, rapid and accurate for determination of chlorogenic acid and can be used for the rapid control of aqueous solution of Flos Lonicerae Japonicae extraction.

HPLC and LC-MS/MS were used to establish a comprehensive HPLC analytical method of Yinqiao decoction and identify the chemical constituents of the whole and individual herbs of Yinqiao decoction. YWG-C18 (250 mm x4. 6 mm ID, 10 microm) column was used; the mobile phase was composed of acetonitrile (A) and water ( B, with 3% acetic acid) with gradient elution; the flow rate was 1. 0 mL x min(-1) and the column temperature was set up at 25 degrees C. The detection wavelength was 280 nm. The chromatographic fingerprints of Yinqiao Decoction showed 30 main peaks. Peak 2, 14, 15, 17 were from Lonicera japonica Thunb, peak 3, 12, 13, 24 were from Fosythia suspense (Thunb) Vahl, peak 19, 25, 26, 27 were from Arctium lappa L. , peak 5, 6, 8, 9, 10, 11, 18, 28 were from Glycyrrhiza uralensis Fisch, peak 20, 21 were from Mentha haplocalyx Briq. , peak 22, 23 were from Schizonepeta tenuifolia Briq. , peak 1 presented in the chromatograms of Lonicera japonica Thunb, Fosythia suspense (Thunb) Vahl, Mentha haplocalyx Briq. , Schizonepeta tenuifolia Briq. and Platycodon grandiflorum (Jacq. ) A. DC. , peak 7 presented in the chromatograms of Fosythia suspense (Thunb) Vahl and Glycine max (L. ) Merr. , peak 16 presented in the chromatograms of Mentha haplocalyx Briq. and Schizonepeta tenuifolia Briq. , peak 29 presented in the chromatograms of the herbs except Mentha haplocalyx Briq. and Platycodon grandiflorum (Jacq. ) A. DC. , peak 30 presented in the chromatograms of the herbs except Platycodon grandiflorum (Jacq. ) A. DC. , peak 4 was not identified, maybe it was a new constituent produced during decoction. By comparison of the standards isolated and MS spectra, 14 peaks were identified as 2 ( chlorogenic acid) , 9 ( liquiritin ) , 10 ( 4'-O-[ beta-D-apiofuranosyl (1--> 2 ) -beta-D-glucopyranosyl] liquiritigenin), 12 (forsythiaside), 13 (rutin), 14 (4,5-O-dicaffeoylquiniic acid), 15 (3, 5-O-dicaffeoylquiniic acid ), 16 ( 4-0- [ beta-D-apiofuranosyl ( 1 -->2 ) -beta-D-glucopyranosyl ] isoliquiritigenin) , 17 ( 3, 4-O-dicaffeoylquiniic acid) , 18 (2'-O-[ beta-D-apiofuranosyl (1 -->2 ) -beta-D-glucopyranosyl] isoliquiritigenin) , 19 (arctiin) , 20 (linarin) , 25 (genistein) , 28 ( isoliquiritigenin) . The method could be used to identify the characteristics of Yinqiao decoction, and it could be used to evaluate the quality and quantity of Yinqiao decoction.

The capillary electrophoresis fingerprints (CEFP) of Flos Lonicerae Japonicae was established to control its quality. In the capillary zone electrophoresis (CZE) pattern 50 mmol/L sodium borate contained 20 mmol/L beta-cyclodextrin (beta-CD) adjusted to pH 8.0 with phosphoric acid was applied as the background electrolyte. The running voltage was 12 kV and the detection wavelength was 254 nm. The Flos Lonicerae Japonicae was extracted by water and the sample solution was injected into the capillary by hydraulic pressure in 15 s. The 18 common peaks were marked according to the emerging rate of 100% and by comparing each fingerprint with the other one among the 13 samples cultivated in different places, in which chlorogenic acid was selected as the reference peak. The CEFP had good precision and reproducibility with the relative standard deviations (RSDs) of the relative migration times less than 3% and the RSDs of the relative peak areas within 6.1%. The novel concepts of the apparent quantitative similarity (R), the quantitative similarity calculated by vector projection (C) and the quantitative similarity (P) were introduced firstly. The good crude drugs should posses two merits as follows: the qualitative similarity (S) which displays the distribution of the chemical constituents in sample should be more than 0.90; the quantitative similarities (R, C, P, Q) that disclose the overall contents of the chemical constituents in sample should be within 80% - 120%. This method was applied in the quality control practice, and the results showed that the method could be used for the overall quality control of Flos Lonicerae Japonicae and is especially suitable for qualitative and quantitative evaluation of chromatographic fingerprints both in chemical constituent distribution and in contents.

A method based on capillary electrophoresis with amperometric detection has been developed for the determination of luteolin, chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeic acid in the dried flower buds, leaves and stems (three medicinal parts) of Lonicera confusa DC., respectively. The effects of several important factors such as detection potential, the concentration of the running buffer, separation voltage and injection time were investigated to acquire the optimum conditions. The detection electrode was a 300 microm diameter carbon disc electrode at a working potential of + 0.90 V (vs saturated calomel electrode). The four analytes can be well separated within 10 min in a 40 cm-long fused silica capillary at a separation voltage of 12 kV in a 50 mM borate-25 mM phosphate buffer (pH 8.0). The relationship between peak current and analyte concentration was linear over about 3 orders of magnitude with detection limits (S/N = 3) ranging from 0.35 to 0.52 microM for all analytes. The proposed method has been successfully applied to the monitoring of bioactive constituents in the real plant samples with satisfactory assay results., (Copyright (c) 2006 John Wiley & Sons, Ltd.)

Objective: To establish an analysical method for the quantitative determination of 18 kinds of organophosphate pesticide residues in Flos Lonicerae., Method: The pesticides in samples were extracted with acetone by Ultrasonic wave Assistant Extraction (UAE), purified by Solid-Phase Extraction (SPE). Using PFPD detecter gas chromatography with HP-1701 column., Result and Conclusion: All 18 pesticides were separated well in 30 minutes. The detection limits was 4.0-18.0 microg x kg(-1). The range of recovery and RSD were 83.64%-88.65% and 2.8%-6.0%, respectively. And it met the requirements of pesticide multi-residues analysis.

The chromatographic fingerprint of Chinese traditional medicine, Yinhuang Oral Liquid together with its original materials Radix Scutellariae and Flos lonicera was established by high performance liquid chromatography (HPLC). At the same time, baicalin and chlorogenic acid in the Yinhuang Oral Liquid were also quantitatively determined. The analysis was performed on a Lichrospher C18 column (250 mm x 4.6 mm i. d. , 5 microm) and with gradient elution. The optimum conditions were explored. The mobile phase A was methanol and the mobile phase B was 0.1% H3PO4 aqueous solution. The wavelength of 254 nm was selected for detection. Finally, the data of fingerprints of 6 samples of Yinhuang Oral Liquid were processed with two different mathematic methods. This proposed method demonstrated good stability for the 6 samples of Yinhuang Oral Liquid. In the comparison of the common peaks in chromatogram of Yinhuang Oral Liquid with those of crude herbs, most of them could be matched well.

Objective: Effects of water stress on the growth of Lonicera japonica and quality of honeysuckle were studied., Methods: Different extent water stress treatment imposed to potted plants before the first florescence of honeysuckle, and the growth and related physiological indexes of Lonicera japonica were determined., Results: Shoot grow slowly as well as the increases of specific weight of leaf and the decrease of the yield of honeysuckle under water stress. The contents of chlorophyl in the leaves treated water stress all increased, and the content of soluble suger had a rising stream. The content of soluble protein in leaves of plant imposed by minor stress treatment was obviously higher than that of CK and severe stress treatment. As the stress was hard, the content of proline in leaves increased significantly. With water stress severing, the content of chlorogenic acid in the flower bud decreased very significantly than CK., Conclusion: Water condition was a important factor affected the growth and development of shoots, leaves and flower buds as well as the quality of honeysuckle. In the cultivation of Loniccra japonica, irrigation timing and certain irrigation condition was necessary.

To establish a rapid and accurate on-line quality control method for aqueous solution of Flos Lonicerae Japonicae extraction (ASFLJE) in the preparation of Qingkailing injection, the wavelength of UV original spectrum and its first derivative spectrum of ASFLJE were scanned. The most suitable spectrum type and wavelength correlated with the chlorogenic acid content in ASFLJE determined by RP-HPLC were chosen to build a regress model. The UV equation for predicting chlorogenic acid content in ASFLJE was established. Other 10 batches of ASFLJE were used for testing the on-line prediction results by comparison of the values of chlorogenic acid content determined by HPLC and predicting equation respectively. The most suitable wavelengths correlated with chlorogenic acid content were 294 nm (r = 0.991 9, n = 28) in the original UV spectrum and 316 nm (r = 0.995 9, n = 28) in the first derivative spectrum respectively. The predicting equation of chlorogenic acid content was: c(mg x mL(-1)) = 506. 254 3 x A316 nm + 0.177 1 (r = 0.995 9, n = 28). This predicting equation is credible and accurate, as tested by HPLC method (r = 0.991 5, n = 10). The prediction results of chlorogenic acid content were: the chlorogenic acid content in probable 90 percent ASFLJE was: 0.4-4.0(mg x mL(-1)). This method is rapid, simple and accurate, and can be used for the on-line quality control of ASFLJE in Qingkailing injection preparation.

Phytochemical analysis is an important scientific research area, which normally relies on a number of rather laborious and time-consuming techniques for compound identification. Isolation of the ingredients of plant extracts in adequate quantities for spectral and biological analysis was the basis of this research. In this paper the possibility of on-line rapid screening of antioxidant components in methanolic plant extracts and their subsequent identification is reported. Based exclusively on hyphenated chromatographic techniques the methanolic extracts of Tilia europea, Urtica dioica, Lonicera periclymenum and Hypericum perforatum are initially screened for their antioxidant components via an on-line DPPH and ABTS radical scavenging technique. Structural elucidation of the active analytes is achieved by means of LC-MS and LC-UV-SPE-NMR. After the determination of the appropriate LC gradient, a minimal number of chromatographic runs with these hyphenated techniques are adequate for the acquisition of the necessary data, leading to the identification of the targeted compounds. Based on their UV, NMR and MS spectra, the antioxidant compounds identified in the extracts under study were found to be either flavonoid glycosides or mono- and dicaffeoylquinic acids. Although the aim of the study was to show the great potential of the LC-UV-NMR-DPPH/ABTS approach for the rapid screening and identification of plant constituents, the results produced in the course of this study also have some merit by themselves. Some of the compounds detected are reported for the first time in the specific plant extracts.

Ochnaflavone is a medicinal herbal product isolated from Lonicera japonica that inhibits cyclooxygenase-2 (COX-2) dependent phases of prostaglandin D2 (PGD2) generation in bone marrow-derived mast cells (BMMC) in a concentration-dependent manner with IC50 values of 0.6 microM. Western blotting probed with specific anti-COX-2 antibodies showed that the decrease in quantity of the PGD2 product was accompanied by a decrease in the COX-2 protein level. In addition, this compound consistently inhibited the production of leukotriene C4 (LTC4) in a dose dependent manner, with an IC50 value of 6.56 microM. These results demonstrate that ochnaflavone has a dual cyclooxygenase-2/5-lipoxygenase inhibitory activity. Furthermore, this compound strongly inhibited degranulation reaction in a dose dependent manner, with an IC50 value of 3.01 microM. Therefore, this compound might provide a basis for novel anti-inflammatory drugs.

A hyphenated method for the isolation and identification of components in a traditional Chinese medicine of Honeysuckle was developed. Ion-exchange chromatography (IEC) was chosen for the fractionation of Honeysuckle extract, and then followed by concentration of all the fractions with rotary vacuum evaporator. Each of the enriched fractions was then further analyzed by reversed-phase liquid chromatography-atmospheric pressure chemical ionization mass spectrometer (RPLC-APCI/MS) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) with matrix of oxidized carbon nanotubes, respectively. It can be noted totally more than 117 components were detected by UV detector, APCI/MS and MALDI-TOF/MS in Honeysuckle extract except the 145 components identified by MALDI-TOF/MS alone with this integrated approach, and 7 of them were preliminary identified according to their UV spectra and mass spectra performed by APCI/MS and MALDI-TOF/MS, respectively. The obtained analytical results not only indicated the approach of integration IEC fractionation with RPLC-APCI/MS and MALDI-TOF/MS is capable of analyzing complex samples, but also exhibited the potential power of the mass spectrometer in detection of low-mass compounds, such as traditional Chinese medicines (TCMs) and complex biological samples.

A reversed-phase high-performance liquid chromatographic (RP-HPLC) method was described for the determination of chlorogenic acid (CGA) in rat plasma using protocatechuic acid as internal standard (IS). CGA in plasma was extracted with acetonitrile, which also acted as deproteinization agent. Chromatographic separation was performed on a Kromasil C18 column with methanol-0.2 m acetic acid (pH 3.0, 25:75, v/v) as mobile phase at a flow-rate of 1.0 mL/min with an operating temperature of 30 degrees C and UV detection at 300 nm. The standard curve was found to be linear over the concentration ranges of 0.4-2.5 microg/mL and 2.5-40 microg/mL, and the limit of quantification (LOQ) was 0.4 microg/mL. The analytical precision and accuracy were validated by relative standard deviation (RSD) and relative error, which were in ranges 3.14-10.78% and -2.20-5.00%, respectively. The average recovery of CGA was 87.59%. The method was successfully applied to the pharmacokinetic study of CGA in Yin-Huang granules., (Copyright (c) 2005 John Wiley & Sons, Ltd.)

Objective: To determine the parameters of pharmacokinetics in rabbits, and to provide a scientific basis for clinical application of Yinhuang compound microenema., Method: HPLC was used to determine the concentrations of chlorogenic acid and baicalin in the plasma samples of rabbits, then analyze these data by the pharmacokinetics program 3P97 to determine the parameters of pharmacokinetics and the compartmental model of both ingredients in Yinhuang compound microenema., Result: Baicalin showed two compartment model in rabbits and the main parameters of pharmacokinetics were as follows: t(1/2)alpha of 0.311 h, t(1/2)beta of 1.640 h, AUC of 1.531 x 10(-2) g x h x L(-1), tmax of 0.287 h, Vd of 2.182 L. Chlorogenic acid showed two compartment model in rabbits and the main parameters of pharmacokinetics were as follows: t(1/2)alpha of 0.324 h, t(1/2)beta of 1.206 h, AUC of 2.135 x 10(-2) g x h x L(-1), tmax of 0.317 h,Vd of 2.251 L., Conclusion: The main components in Yinhuang compound microenema baicalin and chlorogenic acid are absorbed quickly and absolutely after rectal administration, which have good characters of pharmacokinetics.

Objective: To explore the effects of traditional Tibetan medicine, Fructus Lonicerae microphyllae (FLM) on phagecytosis and cytokines production of murine macrophages., Method: The phagecytosis of murine macrophages was analyzed by neutral red phagecytosis assay. The activities of IL-1 and TNF-alpha were measured by biological methods. The mRNA of TNF-alpha and INF-gamma expressed by macrophages was detected by RT-PCR., Result: The phagecytosis of murine macrophages was significantly enhanced by FLM at a concentration from 1 microg x mL(-1) to 100 microg x mL(-1) and the secretions of IL-1, and TNF-alpha from macrophages were markedly induced by FLM. Meanwhile, FLM also increased the expression of TNF-alpha mRNA and INF-gamma mRHA from macrophages in vitro., Conclusion: FLM could promote phagecytosis and cytokines production of murine macrophages.