Chemical compositions and biological activities of Flos lonicerae

Flos lonicerae is the dry flower bud of Lonicera japonica Thunb,and it is a very important Chinese herbal medicine in China.In ancient times,it was widely used to treat the exopathogenic wind-heat,epidemic febrile diseases,sores,carbuncles and some infectious diseases.Modern pharmacological research has indicated that it has anti-inflammatory,anti-bacterial,anti-viral and anti-oxidation activities.This paper reviews the chemical compositions and biological activities of Flos lonicerae in order to provide reference for its application in clinical practice in the future.

Short-term Prediction of Occurrence Period of Main Diseases and Pests of Flos lonicerae using Phenoecology

The occurrence periods of Semiaphis heraclei Takahashi,Frankliniella sp.,Haptonchus luteolus and Microsphara linicerae Enchson wint.in Rabenh.causing damage on Flos lonicerae were investigated in F.lonicerae planting area in XinCheng county of Guangxi Province during 2008-2010,which were coincided with the occurrence periods of related phenology of local Prunus persica Rootstock.With P.persica Rootstock as indicator plant,the occurrence periods of three species of pests and one species of disease were predicted,respectively,and the method was simple and accurate,which could be the foundation for preventing these pests and diseases in the local field.

Separation and Purification of Macranthoidin B and Dipsacoside B from Flos Lonicerae by HP-20 and HP-SS Macroporous Resin

[Objective] This study was conducted to develop a method for rapidly separating macranthoidin B and dipsacoside B from Flos Lonicerae. [Method] HP-20 and HP-SS macroporous resin were applied to separate and purify macranthoidin B and dipsacoside B from Flos Lonicerae. The extract of Flos Lonicerae was first loaded onto an HP-20 column to enrich saponins, which were then separated by an HP-SS macroporous resin column to get pure macranthoidin B and dipsacoside B.[Result] The optimal HP-20 purification conditions included： a concentration of sample liquid at 4.8 mg/ml, a sample volume of 2 BV, an adsorption flow rate at 1.5BV/h, an ethanol concentration for desorption at 60%, a desorption volume of 3 BV,and a desorption flow rate at 1.5 BV/h. Total saponins were then separated by an HP-SS macroporous resin column which was eluted sequentially by water, 20%ethanol, 30% ethanol, 40% ethanol and 50% ethanol. Two purified compounds were obtained in fractions eluted by 40% ethanol and 50% ethanol, respectively. The two compounds were identified as macranthoidin B and dipsacoside B by13 C and1H nuclear magnetic resonance spectroscopy. [Conclusion] The combination of HP-20 and HP-SS macroporous resin could efficiently separate macranthoidin B and dipsacoside B from Flos Lonicerae.

Effect of chlorogenic acid on antioxidant activity of Flos Lonicerae extracts

Flos Lonicerae is a medically useful traditional Chinese medicine herb. However, little is known about the antioxidant properties of Flos Lonicerae extracts. Here the antioxidant capacity of water, methanolic and ethanolic extracts prepared from Flos Lonicerae to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and reduce Fe3+ to Fe2+ is examined. Chlorogenic acid, a major component of Flos Lonicerae, is identified and further purified from 70% ethanolic extract with high performance liquid chromatography (HPLC) and its antioxidant capacity is characterized. The total phenolic compounds and chlorogenic acid con-tents in Flos Lonicerae are determined. The present results demonstrate that the Flos Lonicerae extracts exhibit antioxidant ac-tivity and chlorogenic acid is a major contributor to this activity.

Determination of Chlorogenic Acid,Macranthoidin B and Dipsacoside B in Flos Lonicera by HPLC-ELSD

[Objective] A method for the determination of chlorogenic acid, macranthoidin B and dipsacoside B in Flos Lonicera by high performance liquid chromatography（HPLC） with evaporative light scattering detection（ELSD） was established.[Method] The separation was carried out using a ZORBAX SB-C18 chromatographic column（4.6 mm ×250 mm, 5 μm）, with a simple mobile phase of acetonitrile and0.1% formic acid. Evaporative light scattering detector was employed. [Result] The result showed that chlorogenic aicd, macranthoidin B and dipsacoside B showed good linearities in the ranges of 3.00-6.40, 4.24-9.33 and 0.40-2.00 μg with correlation coefficient of 0.999, 0.993 and 0.999, respectively. [Conclusion] The method has the advantages of simple operation, good accuracy, repeatability and separation effect and high specificity and is suitable for overall quality control of Flos Lonicera.

Comparative analysis of volatile constituents between herbal pair flos lonicerae-caulis lonicerae and its single herbs

Gas chromatography-mass spectrometry(GC-MS) and the chemometric resolution method(alternative moving window factor analysis,AMWFA) were used for comparative analysis of volatile constituents in herbal pair(HP) flos lonicerae-caulis lonicerae(FL-CL) and its single herbs.The temperature-programmed retention index(PTRI) was also employed for the identification of compounds.In total,44,39,and 50 volatile chemical components in volatile oil of FL,CL and HP FL-CL were separately determined qualitatively and quantitatively,accounting for 87.22%,94.54% and 90.08% total contents of volatile oil of FL,CL and HP FL-CL,respectively.The results show that there are 32 common volatile constituents between HP FL-CL and single herb FL,33 common volatile constituents between HP FL-CL and single herb CL,and 10 new constituents in the volatile oil of HP FL-CL.

Screening hepatoprotective effective components of Lonicerae japonica Flos based on the spectrum-effect relationship and its mechanism exploring

Lonicerae japonicae Flos(LF)is a kind of healthcare food with hepatoprotective function.This study was designed to explore the spectrum-effect relationships between UPLC fingerprints and the hepatoprotective effects of LF.Fingerprints of ten batches of LF were established by UPLC-PDA.The inhibitory levels of AST and ALT were used as pharmacological indexes,and secoxyloganin,isochlorogenic acid A and isochlorogenic acid C were screened as hepatoprotective active compounds by grey relational analysis(GRA)and partial least squares regression analysis(PLSR).Caspase-3 was obtained by network pharmacology as a key target of hepatoprotective active compounds.Molecular docking is used to explore the interaction between small molecules and proteins.This work provided a general model of the combination of UPLC-PDA and hepatoprotective effect to study the spectrum-effect relationship of LF,which can be used to considerable methods and insight for the fundamental research of the material basis of similar healthcare food.

Lonicerae Flos:A Review of Chemical Constituents and Biological Activities

Lonicerae flos,a widely used traditional Chinese medicine(TCM),has been used for several thousand years in China.As a famous traditional Chinese herbal medicine,it was used as heat-clearing drug and alexipharmic agent and was widely cultivated in Hunan province.L.flos mainly contains biologically active compounds such as caffeic acid derivatives,essential oil,flavonoids,iridoid glycosides and terpenoids.A range of biological activities has been reported from plant extracts including anti-inflammatory,antitumor,antioxidant,antiallergy,immunomodulating and antibacterial activity.In this study,the author investigated ancient books of TCM and nowadays reports,summarized the chemical constituents,biological activity of L.flos to provide a comprehensive systematic review.

Application of near-infrared spectroscopy for the rapid analysis of Lonicerae Japonicae Flos solution extracted by water

A rapid quantitative analytical method for three components of Lonicerae Japornicae Flos solution(Lonicera Japonica Thumb.)extracted by water was developed using near-infrared(NIR)spectroscopy and the partial least-squares(PLS)method.The NIR spectra of 81 samples collected from a production line were obtained.The concentrations of secologanic acid,chlorogenicacid and galuteolin were detemmined by using high-performance liquid chromatography-diodearray detection as the reference method.Several pretreatment methods for the NIR spectra wereusedi during PLS calibration.The most appropriate latent variable number of the PLS factor wasselected based on the standard error of cross-validation(SECV).The performance of the finalPLS models was evaluated according to SECV,standard error of predliction(SEP)and deter-mination coeficient(R^(2)).The compounds secologanic acid,chlorogenic acid and galuteolin hadSEP values of 0.030,0.061 and 1.668μg/mL,respectively and R^(2) values over 0.85.This workshows that NIR spectroscopy is a rapid and convenient method for the analysis of LoniceraeJaponicae Flos solution extracted by water.The proposed method can help in the application ofprocs analytical technology in the pha maceutical industry,particularly in tra ditional Chinesemedicine injections.

Comparative Analysis of the Complete Chloroplast Genome Sequences of Four Origin Plants of Lonicerae Flos(Lonicera;Caprifoliaceae)

Lonicerae Flos(LF)derived from the dried flower buds or opening flowers of four Lonicera plants(Lonicera macranthoides,L.hypoglauca,L.confusa,and L.fulvotnetosa),is a popular traditional Chinese medicine.Because the four origin plants are very similar in morphology,it is difficult to control the quality of LF in actual production.Over the past decade,many reports have pointed out the differences among them,including the botanical characteristics and active ingredients.However,there is still a lack of rapid methods that can be applied to the identification of the four origins.In this study,comparative analysis of the four chloroplast genomes was performed,and they showed low diversity(Pi=0.00267),three variation hotspots regions(rbcL-accD,rps12-ndhF and rps12-trnN-trnG)were identified as potentially molecular marker of highly informative.Meanwhile,the most obvious difference in SSR comparative analysis is reverse and complement repeats were only identified in L.confusa and L.hypoglauca,respectively.Lastly,the phylogenetic tree showed that L.confusa is more closely related to L.fulvotnetosa,while L.macranthoides is closer to L.hypoglauca.This study systematically revealed the differences among the four chloroplast genomes,and it provides valuable genetic information for identifying the origin of LF.

Leaf epidermal characters of Lonicera japonica and Lonicera confuse and their ecology adaptation

The leaf epidermis of Japanese honeysuckle （Lonicera japonica Thunb.） and Wild Honeysuckle （Lonicera confusa） in the genus of Flos Lonicerae were mainly observed by scanning electron microscopes （SEM） to study the characteristics of stomata, trichomes and dermal cell, etc.. The results showed that stoma exists only on the lower epidermis and its distribution is irregular, and leaf epidermis consist of epidermis cells, stoma complexes and bushy trichomes including glandular hair and non-glandular hair. On the upper epidermis, anticlinal wall caves in sinuous groove to countercheck the transpiration. Evidences from leaf morphological structures serve as another proof on drought-resistant mechanisms. Some strumaes distributing regularly are hypothesized as oxalic calcium on the lower epidermis under laser scanning confocal microscopy （LSCM） with Fluo-3/AM, which can increase their endurance to drought stress. Therefore, the above characteristics of Flos Lonicerae can reduce the loss of water and make Japanese honeysuckle and Wild Honeysuckle adapt to the droughty environment at Karst area in southwest China. However, there is some difference of the two species. From the SEM （Scanning Electron Microscopy） result, it is shown that on the upper epidermis, some glandular hair regularly present along the midrib of Japanese honeysuckle, but Wild Honeysuckle has no glandular hair on the upper epidermis, which can verify the relationships of Flos Lonicerae species and provide the significance for classification of Flos Lonicerae.

Extraction Condition Optimization and Identification of Phenols from Lonicerae flos

Polyphenols were obtained from the natural dried Lonicerae flos by ultrasound-assisted extraction with ethanol as the solvent.Single factor experiment and response surface methodology were employed to optimize the extraction conditions.Ultra-performance liquid chromatrography(UPLC)-tandem mass spectrometry(MS/MS)was employed to identify polyphenols based on the plant widely targeted metabolomics database in a qualitative and quantitative manner.The results showed that the optimal extraction conditions for total phenols from Lonicerae flos were ultrasound-assisted extraction with a solid-to-liquid ratio of 10∶1 g/mL and 57%ethanol at 70 W and 60°C for 11 min.The yield of total phenols extracted under the optimal conditions reached 71.08 mg/g.The phenols in Lonicerae flos were mainly chlorogenic acid isomers,and the flavonoids were mainly nobiletin,galuteolin,and homoarbutin.

Simultaneous quantification of ten phenolic acids in Lonicerae Japonicae Flos by HPLC-DAD

A reverse-phase HPLC-DAD method was developed for simultaneous quantification of ten phenolic acids （caffeic acid, chlorogenic acid, neochlorogenic acid, 4-O-caffeoylquinic acid, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid methyl ester, 3,4-O-dicaffeoylquinic acid methyl ester, and 4,5-0- dicaffeoylquinic acid methyl ester） in the dried flower buds of Lonicera japonica Thunb. （Lonicerae Japonicae Flos; LJF）. An optimal sample preparation method was established as 30-min ultrasonication with 100 times 50% （v/w） ethanol aqueous solution based on the orthogonal test results. The chromatographic separation of the ten phenolic acids was achieved with an AQ-C18 column （4.6 mm x 250 mm, 5 p.m） and a gradient elution of acetonitrile, methanol and 0.1% formic acid aqueous solution within 55 rain. All calibration curves showed good linearity （r2〉0.999） within test ranges. The average recoveries were in the range of 98.57%-103.22% with RSD less than 3%. The method developed was accurate, sensitive and reproducible for determination of ten phenolic acids in LJF.

Systematic review for geo-authentic Lonicerae Japonicae Flos

In traditional Chinese medicine, Lonicerae Japonicae Flos is commonly used as anti-inflammatory, antiviral, and antipyretic herbal medicine, and geo-authentic herbs are believed to present the highest quality among all samples from different regions. To discuss the current situation and trend of geo-authentic Lonicerae Japonicae Flos, we searched Chinese Biomedicine Literature Database, Chinese Journal Full-text Database, Chinese Scientific Journal Full-text Database, Cochrane Central Register of Controlled Trials, Wanfang, and PubMed. We investigated all studies up to November 2015 pertaining to quality assessment, discrimination, pharmacological effects, planting or processing, or ecological system of geo-authentic Lonicerae Japonicae Flos. Sixty-five studies mainly discussing about chemical fingerprint, component analysis, planting and processing, discrimination between varieties, ecological system, pharmacological effects, and safety were systematically reviewed. By analyzing these studies, we found that the key points of geo-authentic Lonicerae Japonicae Flos research were quality and application. Further studies should focus on improving the quality by selecting the more superior of all varieties and evaluating clinical effectiveness.

Comparison of anti-inflammatory effects of Lonicerae Japonicae Flos and Lonicerae Flos based on network pharmacology

Objective:In Chinese herbal medicine(CHM)history,Lonicerae Japonicae Flos and Lonicerae Flos were used clinically as one drug,but now they are admitted as two herbal medicines in Chinese Pharmacopoeia(2010 edition).This study used network pharmacology to investigate whether the two can be used interchangeably for the treatment of inflammatory diseases in TCM clinical practice.Methods:Lonicerae Japonicae Flos and Lonicerae Flos were compared in the inflammation mechanism including core targets,Gene Ontology(GO),pathway and principle chemical components by the method of network pharmacology.Results:Lonicerae Japonicae Flos and Lonicerae Flos shared in six targets accounting for 66.7%of the entire core targets and more than half of the GO terms and pathways are similar.Organic acids are dominent compounds responsible for anti-inflammatory effects.Three of the compounds that bind to core targets including luteolin,quercetin and kaempferol,are shared in both herbs.Conclusion:Due to high similarity between Lonicerae Japonicae Flos and Lonicerae Flos,we believe that they can be used interchangeably for the inflammation in clinical treatment.

Plant metabolomics for studying the effect of two insecticides on comprehensive constituents of Lonicerae Japonicae Flos

Pesticides'overuse and misuse have been reported to induce ingredient variations in herbal medicine,which is now gaining attention in the medicinal field as a form of alternative medicine.To date,available studies on pesticide-induced ingredient variations of herbal medicine are limited only on a few compounds and remain most others unexamined.In this study,a plant metabolomics-based strategy was performed to systematically explore the effects of two frequently used insecticides on the comprehensive constituents of Lonicerae Japonicae Flos(LJF),the flower buds of Lonicera japonica Thunb..Field trials were designed on a cultivating plot of L.japonica with controls and treatments of imidacloprid(IMI)and compound flonicamid and acetamiprid(CFA).Unbiased metabolite profiling was conducted by ultra-high performance liquid chromatography/quadrupole-Orbitrap mass spectrometer.After data pretreatment by automatic extraction and screening,a data matrix of metabolite features was submitted for statistical analyses.Consequently,29 metabolic markers,including chlorogenic acids,iridoids and organic acid-glucosides were obtained and characterized.The relative quantitative assay was subsequently performed to monitor their variations across flowering developments.This is the first study that systematically explored the insecticide-induced metabolite variations of LJF while taking into account the inherent variability of flowering development.The results were beneficial for holistic quality assessment of LJF and significant for guiding scientific use of pesticides in the large-scale cultivation.

Chemical profiling of Jinqi Jiangtang tablets by HPLC-ESI-Q-TOF/MS

AIM： To profile the chemical constituents in Jinqi Jiangtang tablets. METHOD： Based on the chromatographic retention behavior, fragmentation patterns of chemical components, and published lit- eratures, a high-performance liquid chromatography coupled with electrospray ionization quadrnpole time-of-flight tandem mass spectrometry （HPLC-ESI-Q-TOF/MS） method was established to characterize and identify components in Jinqi Jiangtang tablets. RESULTS： A total of 52 chemical compounds, including eight iridoid glycosides, seven phenolic acids, twelve alkaloids, six fla- vonoids, and nineteen saponins, were identified in Jinqi Jiangtang tablets. CONCLUSION： The established method could serve as a powerful tool for structural characterization and quality control of this Chinese herbal preparation.