Rapid quantitative analysis of adulterant Lonicera species in preparations of Lonicerae Japonicae Flos

Lonicerae Japonicae Flos is often adulterated with Lonicerae Flos, which is derived from the other four Lonicera species, in both the crude drug and Lonicerae Japonicae Flos preparations. We proposed a methodology for the quantitative analysis of adulterant Lonicerae Flos in Lonicerae Japonicae Flos preparations. Taking macranthoidins A, B, dipsacoside B (saponins), sweroside (iridoids), and luteolin‐7‐O‐d ‐glucoside (flavonoids) as markers, a method of ultra high performance liquid chromatography with triple quadrupole mass spectrometry was employed to determine their amounts in Lonicerae Flos, Lonicerae Japonicae Flos, and Lonicerae Japonicae Flos preparations. The proportion of adulterant Lonicerae Flos in Lonicerae Japonicae Flos preparations was estimated based on the saponin contents of Lonicerae Japonicae Flos and Lonicerae Flos. All analytes separated under isocratic elution in 12 min with acceptable linearity, precision, repeatability, and accuracy. Lonicerae Japonicae Flos was easily distinguished from Lonicerae Flos by the total amount of saponins (0.067 and > 45.8 mg/g for Lonicerae Japonicae Flos and Lonicerae Flos, respectively). Eighteen of twenty one Lonicerae Japonicae Flos preparation samples were adulterated with Lonicerae Flos in proportions of 11.3–100%. The developed ultra high performance liquid chromatography with triple quadrupole mass spectrometry method could be used for the identification of Lonicerae Japonicae Flos and the four species of Lonicerae Flos and for the analysis of Lonicerae Japonicae Flos preparations adulterated with Lonicerae Flos.     

Simultaneous determination of iridoids, phenolic acids, flavonoids, and saponins in Flos Lonicerae and Flos Lonicerae Japonicae by HPLC-DAD-ELSD coupled with principal component analysis

A method, HPLC coupled with diode-array and evaporative light scattering detectors (HPLC-DAD-ELSD), was newly developed to evaluate the quality of Flos Lonicerae (FL) and Flos Lonicerae Japonicae (FLJ), through a simultaneous determination of multiple types of bioactive components. By employing DAD, the detection wavelengths were set at 240 nm for the determination of iridoids, 330 nm for phenolic acids, and 360 nm for flavonoids, respectively. While ELSD, connected in series after DAD, was applied to the determination of saponins. This assay was fully validated with respect to precision, repeatability, and accuracy. Moreover, principal component analysis (PCA) was used for the similarity evaluation of different samples, and it was proven straightforward and reliable to differentiate FL and FLJ samples from different origins. For PCA, two principal components have been extracted. Principal component 1 (PC1) influences the separation between different sample sets, capturing 54.598% variance, while principal component 2 (PC2) affects differentiation within sample sets, capturing 12.579% variance. In conclusion, simultaneous quantification of bioactive components by HPLC-DAD-ELSD coupled with PCA would be a well-acceptable strategy to differentiate the sources and to comprehensively control the quality of the medicinal plants FL and FLJ.     

Capillary high-performance liquid chromatography with mass spectrometry for simultaneous determination of major flavonoids, iridoid glucosides and saponins in Flos Lonicerae

Flos Lonicerae, a traditional herbal medicine, has been used in China to treat some inflammatory disease. Several different classes of compounds have been separated from the herb to assess their pharmacological activities. Among these classes, flavonoids, iridoid glycosides and saponins have been well studied and may be responsible for its clinical application. Therefore, quality control of Flos Lonicerae is an important issue for drug safety and validity evaluations. A quantitative method consisting of solid phase extraction followed by capillary high-performance liquid chromatography-diode array detection/electrospray ionization mass spectrometry (capillary HPLC-DAD/ESI-MS) was developed for simultaneously assay of 24 compounds in Flos Lonicerae. Under optimized capillary HPLC-DAD/ESI-MS conditions, these compounds, including nine flavonoids, eight iridoid glucosides and seven saponins, were separated with high efficiency in the selected-ion monitoring (SIM) mode. Linearity of the method was good with correlation coefficients (r(2)) in the range of 0.9935-0.9998 and detection limits were lower than 2.57 ng/mL for most of analytes. The obtained recoveries varied between 91.0 and 108.7% with the relative standard deviations (RSDs) within 8.74% (n=3). The capillary HPLC-DAD/ESI-MS method was also successfully applied to the analysis of these compounds in five species of Flos Lonicerae. It was demonstrated to be a powerful tool for comprehensive analysis of herbal medicines, owing to its exclusive selectivity and excellent sensitivity.     

Characterization and identification of multiple constituents in Yinhuang granules by high-performance liquid chromatography with diode-array and time-of-flight mass spectrometry detection

A fast high-performance liquid chromatography (HPLC) method with diode-array detection (DAD) and time-of-flight mass spectrometry (TOF/MS) has been developed for the analysis of multi-constituent in Yinhuang granules, a well-known combined herbal remedy prepared from the extract mixtures of Flos Lonicerae and Radix Scutellariae. The fast HPLC analysis was performed on an Agilent ZorBax SB-C(18) column (4.6×50 mm, 1.8 μm) and 0.2% aqueous formic acid and acetonitrile was the optimum mobile phase for gradient elution in 17 min, which is five times faster than the performance of conventional columns packed with 5.0 μm particles. With various fragmentor voltages in TOF/MS, accurate mass measurements (<5 ppm error) for molecular ions and characteristic fragment ions represented reliable identification criteria for different constituents. A total of 28 compounds, including nine phenolic acids, three iridoid glycosides and nine saponins from Flos Lonicerae and seven flavonoids from Radix Scutellariae, were identified or tentatively characterized in the extract of Yinhuang granules. The established fast HPLC-DAD-TOF/MS method turns out to be useful and efficient for quality control of this commonly used Chinese herbal preparation.     

微波辅助萃取/高效液相色谱串联质谱法对山银花中活性成分含量的测定

建立了微波辅助萃取/高效液相色谱串联质谱法(MAE/HPLC-MS/MS)同时测定山银花中10种活性成分含量的方法。山银花药材采用MAE萃取,萃取溶剂为乙醇-水(7∶3),固液比1∶30,萃取温度70℃,萃取时间10 min。采用HPLC-MS/MS测定萃取液中活性成分的含量,色谱柱采用Agilent Poroshell120 SB-C18(100 mm×2.1 mm,2.7μm),以0.5%甲酸-乙腈为流动相进行梯度洗脱,负离子多重反应离子监测模式检测。在优化条件下,10种成分的定量分析在10 min内完成。结果表明,10种活性成分的线性范围为0.05～500 mg/L,相关系数(r)不低于0.996 9,检出限和定量下限分别在69～4 413μg/kg和231～14709μg/kg范围,回收率为94%～105%。采用该方法检测6个不同产地的山银花样品,10种活性成分的含量在3.98～14 356.31 mg/kg范围。该方法快速、准确,可有效地用于山银花药材的质量控制。     

Four new saponins from the root bark of Aralia elata

Four new saponins, 3-O-[beta-D-glucopyranosyl(1-->3)-alpha-L-arabinopyranosyl]-16a lpha-hydroxyoleanolic acid 28-O-beta-D-glucopyranosyl ester (called aralia-saponin I), 3-O-[beta-D-glucopyranosyl(1-->3)-alpha-L-arabinopyranosyl]-16a lpha-hydroxyhederagenin 28-O-beta-D-glucopyranosyl ester (aralia-saponin II), 3-O-[beta-D-glucopyranosyl(1-->3)-beta-D-glucopyranosyl(1-->3)-alpha-L-+ ++arabinopyranosyl]-16alpha-hydroxyoleanolic acid 28-O-beta-D-glucopyranosyl ester (aralia-saponin III), 3-O-[beta-D-glucopyranosyl(1-->3)-beta-D-gucopyranosyl(1-->3)-beta -D-glucucopyranosyl]-16alpha-hydroxyoleanolic acid 28-O-beta-D-glucopyranosyl ester (aralia-saponin IV), were isolated from the root bark of Aralia elata (Miq.) Seem., together with nineteen known compounds including glycosides of (20S)-protopanaxadiol and (20S)-protopanaxatriol. Their structures were determined on the basis of chemical and spectroscopy methods.     

Two new triterpenoid saponins isolated from Polygala japonica

Bioassay guided investigation of whole parts of Polygala japonica afforded two new triterpenoid saponins, characterized as 3-O-beta-D-glucopyranosyl medicagenic acid 28-O-{beta-D-xylopyranosyl(1-->4)-[beta-D-apiofuranosyl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl} ester (1), 3-O-beta-D-glucopyranosyl 2-oxo-olean-12-en-23, 28-dioic acid 28-O-{beta-D-xylopyranosyl(1-->4)-[beta-D-apiofuranosyl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl} ester (2), together with four known triterpenoid saponins (3-6). Their structures were elucidated by spectroscopic and chemical methods. Saponins 3, 4 and 5 showed significant anti-inflammation effects on carrageenan-induced acute paw edema in mouse.     

Saponins and other constituents from the leaves of Aralia elata

A new triterpenoid saponin, together with five known saponins, were isolated from the nonpolar n-hexane fraction of the leaves of Aralia elata. The structure of the new saponin, durupcoside C, was elucidated as hederagenin 3-O-beta-D-glucopyranosyl(1-->3)-beta-D-glucopyranosyl(1-->3)-alpha-L-arabinopyranoside on the basis of spectroscopic analysis. The known saponins were characterized as 3-O-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-xylopyranosyl(1-->6)-beta-D-glucopyranosyl ester, hederagenin 3-O-beta-D-glucopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside, oleanolic acid 3-O-beta-D-glucopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside, hederagenin 3-O-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside (alpha-hederin), and hederagenin 3-O-beta-D-glucopyranosyl(1-->3)-alpha-L-arabinopyranoside (collinsonidin). In addition, two known lipids, Arisaema glyceride 3 and ceramide mixtures were also isolated and characterized. Collinsonidin and two known lipids were isolated for the first time from this plant.     

New triterpenoid saponins from Stelmatocrypton khasianum

Four new triterpenoid saponins, designated as stelmatotriterpenosides E-H (1-4), together with three known compounds, asterbatanoside B (5), 2alpha,3beta,19alpha,23-tetrahydroxy-olean-12-en-28-oic acid-3-O-beta-D-glucopyranosyl-28-O-beta-D-glucopyranosyl ester (6) and 2alpha,3beta,19alpha,23-tetrahydroxy-urs-12-en-28-oic acid-3-O-beta-D-glucopyranosyl-28-O-beta-D-glucopyranosyl ester (7), were isolated from the stems of Stelmatocrypton khasianum. On the basis of chemical and spectral evidence, the structures of 1-4 were established as 2alpha,3beta,23-trihydroxy-olean-12-en-28-oic acid-3-O-beta-D-glucopyranosyl-28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester (1), 2alpha,3beta,23-trihydroxy-urs-12-en-28-oic acid-3-O-beta-D-glucopyranosyl-28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester (2), 2alpha,3beta,19alpha-trihydroxy-urs-12-en-28-oic acid-3-O-beta-D-glucopyranosyl-28-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl ester (3), and 2beta,3beta,19alpha-trihydroxy-urs-12-en-24,28-dioic acid-24-O-beta-D-glucopyranosyl-28-O-beta-D-glucopyranosyl diester (4).     

New A-ring lactone triterpenoid saponins from the roots of Platycodon grandiflorum

Three new A-ring lactone triterpenoid saponins, platycoside M-1 [3-O-beta-D-glucopyranosyl platycogenic acid A lactone], platycoside M-2 [3-O-beta-D-glucopyranosyl platycogenic acid A lactone 28-O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside], and platycoside M-3 [3-O-beta-D-glucopyranosyl platycogenic acid A lactone 28-O-beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside], were isolated from the roots of Platycodon grandiflorum A. DC. Their chemical structures were elucidated on the basis of their spectral data and chemical evidence.     

Two new cycloartane saponins from the roots of Astragalus membranaceus

Two new cycloartane-type triterpenoid saponins were isolated from the roots of Astragalus membranaceus (FISCH.) BGE. (Leguminosae) cultivated in Kangwon province, Korea. These saponins were named astramembranosides A and B and were established to be cycloastragenol 6,25-di-O-beta-D-glucopyranoside (astramembranoside A) and cyclocanthogenin 3-O-beta-D-glucopyranosyl(1-->2)-beta-D-xylopyranoside (astramembranoside B) on the basis of chemical and spectral evidence. In addition, 12 known saponins were also isolated from the same materials. Although cycloastragenol 3-O-xyloside and agroastragalosides I and II have already been isolated from A. membranaceus adventitious roots, these three saponins together with brachyoside B and azukisaponin V methyl ester were isolated for the first time from this plant.     

Latifolosides K and L, two new triterpenoid saponins from the bark of Llex latifolia.

Two new triterpenoid saponins, latifoloside K (1), 3-O-beta-D-glucopyranosyl-(1-->3)-[alpha-L-rhamnopyranosyl-(1-->2)-]-alpha-L-arabinopyranosyl 3beta-hydroxy-urs-12,18-dien-28-oic acid 28-O-beta-D-glucopyranosyl ester and latifoloside L (2), 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->3)-[alpha-L-rhamnopyranosyl-(1-->2)-]-alpha-L-arabinopyranosyl 3beta,19alpha-dihydroxyursolic acid, were isolated from the bark of Ilex latifolia Thunb. Also isolated were two known compounds, ilekudinoside A (3) and kudinoside G (4). Structural assignments were established on the basis of spectroscopic data and chemical evidence.     

Saponins from leaves of Acanthopanax hypoleucus Makino.

From the leaves of Acanthopanax hypoleucus Makino (Araliaceae), five triterpenoidal saponins, having oleanolic acid and hederagenin as sapogenins, were isolated. On the basis of chemical and spectral data, the structures of two new saponins, named hypoleucosides A (1), and B (5) were elucidated as follows: 1; 3-O-beta-D-glucopyranosyl 11 alpha-methoxy-oleanolic acid 28-O-beta-D-glucopyranosyl ester, 5; 3-O-beta-D-glucopyranosyl-(1----2)-alpha-L-arabinopyranosyl-(1---- 4)-beta-D-glucopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1----6)-beta-D-glucopyranosyl ester.     

Bacopasides III-V: three new triterpenoid glycosides from Bacopa monniera

Three new saponins, designated as bacopasides III, IV and V have been isolated from Bacopa monniera WETTST. and their structures have been elucidated as 3-O-alpha-L-arabinofuranosyl (1-->2)-beta-D-glucopyranosyl jujubogenin (1), 3-O-beta-D-glucopyranosyl (1-->3)-alpha-L-arabinopyranosyl jujubogenin (2) and 3-O-beta-D-glucopyranosyl (1-->3)-alpha-L-arabinopyranosyl pseudojujubogenin (3) mainly on the basis of two dimensional (2D) NMR and other spectral analyses.     

Determination of five major iridoid glucosides in Flos Lonicerae by high-performance liquid chromatography coupled with evaporative light scattering detection

This study presents a new HPLC method using evaporative light scattering detection for the simultaneous determination of live major iridoid glucosides, namely 7-epi-loganin, sweroside, loganin, 7-epi-vogeloside, and secoxyloganin in Flos Lonicerae, an important traditional Chinese medicinal herb. The optimal conditions of separation and detection were achieved on a C18 analytical column with an isocratic mobile phase consisting of methanol-water (30:70, v/v) containing 0.5% acetic acid at the flow-rate of 1.0 ml/min, temperature for the detector drift tube set at 90 degrees C and the nitrogen flow-rate of 2.6 l/min. The limit of detection (S/N = 3) is less than 35.1 microg/ml and the limit of quantification (S/N = 10) is less than 140.1 microg/ml. All calibration curves show good linear regression (r2>0.996) within test ranges. This method provides good reproducibility for the quantification of the major iridoid glucosides in four Lonicera species with overall intra- and inter-day variation of less than 5% and 9%, respectively. The assay was successfully applied to quantify the main iridoid glucosides in the herb and to identify the botanical origin of Flos Lonicerae.     

Bioactive saponins and glycosides. XVI. Nortriterpene oligoglycosides with gastroprotective activity from the fresh leaves of Euptelea polyandra Sieb. et Zucc. (1): structures of Eupteleasaponins I, II, III, IV, V, and V acetate

The saponin fraction from the fresh leaves of Euiptelea polyandra Sieb. et Zucc. was found to exhibit potent gastroprotective activity. Fourteen new nortriterpene saponins called eupteleasaponins were isolated from the saponin fraction with gastroprotective activity. The structures of eupteleasaponins I, I1, III, IV, V, and V acetate were determined on the basis of chemical and physicochemical evidence as 3-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl(1 -->3)-beta-D-xylopyranosylakebonoic acid 28-O-beta-D-glucopyranosyl ester, 3-O-alpha-L-rhamnopyranosyl(1-->2)-[alpha-L-rhamnopyranosyl(1 -->4)-beta-D-glucopyranosyl(1-->3)-beta-D-xylopyranosylakebonoic acid 28-O-beta-D- D-glucopyranosyl(1-->3)-beta-D-xylopyranosylakebonoic acid 28-O-beta-D-glucopyranosyl ester, 3-O-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-rhamnopyranosyl(1 -->4)-beta-D-glucopyranosyl(1-->3)-beta-D-xylopyranosylakebonoic acid 28-O-beta-D-glucopyranosyl ester, 3-O-alpha-L-rhamnopyranosyl(1 -->2)-[alpha-L-arabinopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-- >4)]-beta-D- glucopyranosyl(1-->3)-beta-D-xylopyranosylakebonoic acid 28-O-beta-D-glucopyranosyl ester, 3-O-alpha-L-rhamnopyranosvl(1-->2)-[alpha-L-arabinopyranosyl (1 -->4)-beta-L-rhamnopyranosyl(1 -->4)]-beta-D-glucopyranosyl(1 -->3)-beta-D-xylopyranosylakebonoic acid, 3-O-alpha-L-rhamnopyranosyl(1 -->2)-beta-D-glucopyranosvl(1-->3)-beta-D-xylopyranosyleupteleo genin, and 3-O-alpha-L-rhamnopyranosyl(1-->2)-6"-O-acetyl-gamma-D-glucopyranosyl(1 -->3)-beta-D-xylopyranosyleupteleogenin.     

A new triterpenoid saponin from Gleditisia sinensis and structure-activity relationships of inhibitory effects on lipopolysaccharide-induced nitric oxide production

A phytochemical investigation of the anomalous fruits of Gleditisia sinensis led to one new oleanane-type triterpenoid saponin acylated with one monoterpenic acid, 3-O-beta-D-glucopyranosyl oleanolic acid 28-O-beta-D-xylopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-alpha-L-rhamnopyranosyl-(1 --> 2)-[(6S,2E)-6-hydroxy-2,6-dimethyl-2,7-octadienoyl-(1 --> 6)]-beta-D-glucopyranosyl ester, named gleditsioside Z (1), together with nine known ones (2-10). Their structural details were mainly established on the basis of 1D, 2D NMR and HR-MS analysis as well as some chemical methods. Structure-activity relationships of the isolated saponins that affected nitric oxide production from cultured mouse macrophages (RAW 264.7 cell lines) induced by lipopolysaccharide were discussed.     

Simultaneous determination of 11 saponins in Panax notoginseng using HPLC-ELSD and pressurized liquid extraction

A new HPLC coupled with evaporative light scattering detection (ELSD) method was developed for simultaneous determination of 11 major triterpene saponins, namely notoginsenoside R1 (1), ginsenosides Rg1 (2), Re (3), Rf(4), Rb1 (5), Rg2 (6), Rc (7), Rb2 (8), Rb3 (9), Rd (10), and Rg3 (11) in Panax notoginseng, a commonly used traditional Chinese medicine (TCM). Pressurized liquid extraction (PLE) was employed for sample preparation, and the analysis was achieved using a Zorbax ODS C18 column eluted with gradient water-ACN in 60 min. The drift tube temperature of ELSD was set at 60 degrees C, and nitrogen flowrate was at 1.4 L/min. The method provided good repeatability and sensitivity for quantification of 11 saponins with overall precision (including intra- and interday) and LOD of less than 2.9% (RSD) and 98 ng, respectively. The validated method was successfully applied to quantify 11 saponins in 28 samples of P. notoginseng collected in different places, which is helpful to control the quality of P. notoginseng and its related products.     

Analysis of Chlorogenic Acid in Herbal Medicines by Capillary Electrophoresis

Atpresent,thecommonlyusedmethodforanalyzingherbalmedicinesishighperformanceliquidchromatography(HPLC).However,theuseofHPLCisrestrictedformanyreasons.Themostseriousproblemisthatthechromatographiccolumniseasilycontaminated.ComparedwithHPLC,capillaryelectrophoresis(CE)ischaracterizedwithhighefficiency,rapidity,lowcostandmultiplemodestobechosen.Inaddition,thecapillarycanbeeasilyregenerated.SoCEisidealforanalyzing"dirty"samples,suchasChineseherbalmedicines.Infact,studiesintheareaarebecoming…