On the chemical constituents of Dipsacus asper

Bioassay-guided fractionation of 95% EtOH extract from the roots of Dipsacus asper lead to the isolation of some phenolic acids (caffeic acid, 2,6-dihydroxycinnamic acid, vanillic acid, 2'-O-caffeoyl-D-glucopyranoside ester, and caffeoylquinic acid) as the major active components, and five new iridoid glucoside dimers (1-5) and one new iridoid glucoside monomer (6), other known iridoid glycosides loganin, cantleyoside, triplostoside A, lisianthioside, 6'-O-beta-D-apiofuranosyl sweroside, as well as triterpenoids oleanic acid and akebiasaponin D. The structures of new compounds 1-6 were determined as dipsanosides C (1), D (2), E (3), F (4), G (5), and 3'-O-beta-D-glucopyranosyl sweroside (6) by spectroscopic, including 1D and 2D NMR techniques, and chemical methods.     

A new iridoid glycoside from the roots of Dipsacus asper

A new iridoid glycoside, named loganic acid ethyl ester (1), together with five known compounds: chlorogenic acid (2), caffeic acid (3), loganin (4), cantleyoside (5) and syringaresinol-4',4'-O-bis-β-D-glucoside (6) were isolated from the roots of Dipsacus asper. The structure of compound 1 was elucidated on the basis of detailed spectroscopic analyses. Lignan is isolated from Dipsacaceae species for the first time. Compounds 1, 4 and 5 had moderate neuroprotective effects against the Aβ????? induced cell death in PC12 cells.     

Determination of asperosaponin VI in rat plasma by HPLC‐ESI‐MS and its application to preliminary pharmacokinetic studies

Asperosaponin VI (also named akebia saponin D) is a typical bioactive triterpenoid saponin isolated from the rhizome of Dipsacus asper Wall (Dipsacaceae). In this work, a sensitive high‐performance liquid chromatography–electrospray ionization–mass spectrometry (HPLC‐ESI‐MS) assay has been established for determination of asperosaponin VI in rat plasma. With losartan as the internal standard (IS), plasma samples were prepared by protein precipitation with methanol. Chromatographic separation was performed on a C₁₈ column with a mobile phase of 10 mm ammonium acetate buffer containing 0.05% formic acid–methanol (32 : 68, v/v). The analysis was performed on an ESI in the selected ion monitoring mode using target ions at m/z 951.4 for asperosaponin VI and m/z 423.2 for the IS. The calibration curve was linear over the range 3–1000 ng/mL and the lower limit of quantification was 3.0 ng/mL. The intra‐ and inter‐assay variability values were less than 9.5 and 7.8%, respectively. The accuracies determined at the concentrations of 3.0, 100.0, 300.0 and 1000 ng/mL for asperosaponin VI were within ±15.0%. The validated method was successfully applied to a pharmacokinetic study in rats after oral administration of asperosaponin VI. Copyright © 2009 John Wiley & Sons, Ltd.     

Ultra‐high‐performance liquid chromatography with tandem mass spectrometry method for determination of four compounds in rat plasma after oral administration of Xanthii fructus and stir‐fried Xanthii fructus extracts

Xanthii fructus (XF), the fruit of Xanthium sibiricum Patr., is a traditional Chinese materia medica commonly used to treat allergic rhinitis and other rhinitis diseases. To uncover the mechanism of the stir‐frying process and its effect on the pharmacokinetic behavior of active compounds in model rats, four active compounds—chlorogenic acid, 4‐caffeoylquinic acid, 1,5‐O‐dicaffeoylquinic acid and apigenin—were selected based on previous spectrum‐effect experiments. High performance liquid chromatography tandem triple quadrupole mass spectrometry (UPLC–QqQ–MS) technology, an accurate and feasible method, was applied to measure the concentration of these four compounds in rat plasma. This validated method can accurately measure the concentration of each compound at each sampling point of rat plasma. This validated method shows good linearity, extraction recoveries, matrix effects, intra‐ and inter‐day precision and stabilities. Compared with the XF group, the maximum plasma concentration (C_max) value of 1,5‐O‐dicaffeoylquinic acid decreased remarkably (p < 0.05) after oral administration of stir‐fried Xanthii fructus (SXF) extract, while the other compounds showed no significant difference. The mean residence time value of chlorogenic acid (p < 0.05) and 1,5‐O‐dicaffeoylquinic acid (p<0.01) after oral administration of SXF extraction demonstrated significant differences compared with the XF group, while the other two compounds showed no statistical difference, indicating that the stir‐frying process prolonged the effect time and delayed the removal time of chlorogenic acid and 1,5‐O‐dicaffeoylquinic acid. The values of the area under the plasma concentration–time curve from zero to the last quantifiable time‐point, the area under the plasma concentration–time curve from zero to infinity, the time to maximum concentration and the elimination half‐life of four compounds in the SXF group showed no statistically significant difference from the XF group. From this data, we speculated that the stir‐frying process can not only keep the absorption of 4‐caffeoylquinic acid and apigenin, but also increase the effect time of chlorogenic acid and 1,5‐O‐dicaffeoylquinic acid, which could be the mechanism underlying the stir‐frying process enhancing the effects of XF.     

Hierarchical scheme for liquid chromatography/multi‐stage spectrometric identification of 3,4,5‐triacyl chlorogenic acids in green Robusta coffee beans

Liquid chromatography/multi‐stage spectrometry (LC/MSⁿ) (n = 2–4) has been used to detect and characterize in green Robusta coffee beans eight quantitatively minor triacyl chlorogenic acids with seven of them not previously reported in nature. These comprise 3,4,5‐tricaffeoylquinic acid (Mr 678); 3,5‐dicaffeoyl‐4‐feruloylquinic acid, 3‐feruloyl‐4,5‐dicaffeoylquinic acid and 3,4‐dicaffeoyl‐5‐feruloylquinic acid (Mr 692); 3‐caffeoyl‐4,5‐diferuloylquinic acid and 3,4‐diferuloyl‐5‐caffeoylquinic acid (Mr 706); and 3,4‐dicaffeoyl‐5‐sinapoylquinic acid and 3‐sinapoyl‐4,5‐dicaffeoylquinic acid (Mr 722). Structures have been assigned on the basis of LC/MSⁿ patterns of fragmentation. A new hierarchical key for the identification of triacyl quinic acids is presented, based on previously established rules of fragmentation. Fifty‐two chlorogenic acids have now been characterized in green Robusta coffee beans. In this study five samples of green Robusta coffee beans and fifteen samples of Arabica coffee beans were analyzed with triacyl chlorogenic acids only found in Robusta coffee bean extracts. These triacyl chlorogenic acids could be considered as useful phytochemical markers for the identification of Robusta coffee beans. Copyright © 2010 John Wiley & Sons, Ltd.     

A rapid and sensitive UPLC‐MS/MS method for quantification of two caffeoylquinic acids and four main active components in rat plasma after an intravenous administration of Qingkailing injection and its application to a pharmacokinetic study

Qingkailing (QKL) injection, a modified modern Chinese medicine preparation, is widely used in the clinic for its significant antipyretic and anti‐inflammatory effects, but its serious adverse drug reactions have attracted more and more attention. Series of caffeoylquinic acids in QKL are widely suspected to be the allergens responsible for these adverse drug reactions. Therefore, pharmacokinetic studies of the caffeoylquinic acids are needed. In this paper, a simple, rapid and sensitive ultra‐performance liquid chromatography–tandem mass spectrometry method was developed for the simultaneous determination of chlorogenic acid, neochlorogenic acid, baicalin, geniposide, cholic acid and hyodeoxycholic acid in rat plasma. Chromatographic separation was achieved on a BEH C₁₈ column by a gradient elution at a flow rate of 0.40 mL/min in only 6.0 min. All analytes were monitored by multiple reaction monitoring mode with negative electrospray ionization. The calibration curves of these analytes were all linear (r > 0.9978) over wide concentration ranges. The intra‐ and inter‐ day precisions (relative standard deviations) were within 14.3% and accuracy (relative error) ranged from ?6.8 to 4.8%. The mean recoveries ranged from 74.5 to 105.6%. This validated method was successfully applied to the pharmacokinetic study of the six analytes in rats following an intravenous administration of QKL injection. Copyright © 2013 John Wiley & Sons, Ltd.     

Simultaneous separation and determination of phenolic acids,pentapeptides, and triterpenoid saponins in the root of Aster tataricus by high‐performance liquid chromatography coupled with electrospray ionization quadrupole time‐of‐flight mass spectrometry

We established a qualitative method to analyze the main chemical compositions of the root of Aster tataricus. Most of the peaks were separated on a C₁₈ column packed with 5.0 μm particles, and 28 compounds were identified, including 11 chlorogenic acids, ten astins/asterinins, and seven astersaponins, four of which were reported for the first time from A. tataricus. Furthermore, we developed a reliable method for the simultaneous quantification of 3‐caffeoylquinic acid, 3,4‐dicaffeoylquinic acid, 3,5‐dicaffeoylquinic acid, astin A, astin B, astin C, astersaponin A, and astersaponin C, and the qualified separations were achieved only on a C₁₈ column packed with 2.7 μm particles. The method was used to measure the concentrations of eight components in samples from two major producing areas in China, and the average contents in samples from Bozhou (Anhui) were higher than those in samples from Anguo (Hebei).     

Rapid determination of caffeoylquinic acid derivatives in Cynara scolymus L. by ultra‐fast liquid chromatography/tandem mass spectrometry based on a fused core C18 column

An ultra‐fast high‐performance LC‐ESI‐MS/MS method was developed for the analysis and quantification of caffeoylquinic acid derivatives, including chlorogenic acid, 1,3‐di‐O‐caffeoylquinic acid (cynarin) and 1,5‐di‐O‐caffeoylquinic acid, in artichoke (Cynara scolymus L.) heads and leaves. The rapid separation (less than 4 min) was achieved based on a Halo fused core C18‐silica column (50 mm×2.1 mm id, 2.7 μm). The target compounds were detected and quantified by a triple‐quadrupole mass spectrometer in multiple‐reaction monitoring mode. The calibration function is linear from 0.06 to 2800 ng/mL for chlorogenic acid, 0.3–3000 ng/mL for cynarin and 0.24–4800 ng/mL for 1,5‐di‐O‐caffeoylquinic acid, respectively. The average recoveries ranged from 92.1 to 113.2% with RSDs ≤6.5%. Moreover, four batches of artichoke head and leaf extracts were analyzed using the established method. The results indicated that the Halo fused core column provided much faster separations and higher sample throughput without sacrificing column ruggedness and reliability, and triple‐quadrupole MS provided extraordinarily lower LOQs for most of the target analytes. Comparing to conventional quantitative approaches, the established method was fast, sensitive and reliable for the determination of caffeoylquinic acid derivatives in artichoke.     

Comparison of the active components of Aster tataricus from different regions and related processed products by ultra‐high performance liquid chromatography with tandem mass spectrometry

We investigated crude Aster tataricus, vinegar‐processed Aster tataricus, honey‐processed Aster tataricus, and steamed Aster tataricus as a case study and developed a comprehensive strategy integrating quantitative analysis and chemical pattern recognition methods for the evaluation and differentiation of Aster tataricus from different regions, as well as related processed products. In the study, 15 batches of raw Aster tataricus collected from seven provinces were analyzed. A sensitive and rapid ultra‐high performance liquid chromatography with tandem mass spectrometry method for simultaneous determination of 15 compounds was established to evaluate the quality of raw and processed Aster tataricus. Furthermore, multivariate statistical techniques were applied to compare the differences among Aster tataricus samples. As a result, the herbs collected from seven provinces were divided into two categories, and chlorogenic acid was the most important component distinguishing between the regions. Moreover, all of the raw and processed samples were classified by partial least squares discriminant analysis based on the 15 analyzed compounds. Results showed that raw Aster tataricus, vinegar‐processed Aster tataricus, honey‐processed Aster tataricus, and steamed Aster tataricus were clustered in four different areas. Shionone, chlorogenic acid and kaempferol were the significant constituents differentiating the raw and differently processed Aster tataricus samples.     

Simultaneous determination of four secoiridoid and iridoid glycosides in rat plasma by ultra performance liquid chromatography–tandem mass spectrometry and its application to a comparative pharmacokinetic study

A simple, reliable and rapid ultra‐performance liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantification of four secoiridoid (gentiopicroside, swertiamarin, sweroside) and iridoid glycosides (loganic acid), the bio‐active ingredients in rat plasma. After liquid–liquid extraction, chromatographic separation was accomplished on a Shim‐pack XR‐ODS column with a mobile phase consisting of methanol and 0.1% formic acid in water. A triple quadrupole tandem mass spectrometry equipped with an electrospray ionization source was used as detector operating both in positive and negative ionization mode and operated by multiple‐reaction monitoring scanning. The lower limits of quantitation were 0.25–30 ng/mL for all the analytes. Both intra‐day and inter‐day precision and accuracy of analytes were well within acceptance criteria (±15%). The mean extraction recoveries of analytes and internal standard (amygdalin) from rat plasma were all >71.4%. The validated method was successfully applied to a comparative pharmacokinetic study of four analytes in rat plasma between normal and arthritic rats after oral administration of Huo Luo Xiao Ling Dan and Gentiana macrophylla extract, respectively. Results showed significant differences in pharmacokinetic properties of the analytes among the different groups. Copyright © 2015 John Wiley & Sons, Ltd.     

Different fingerprinting strategies to differentiate Porana sinensis and plants of Erycibe by high‐performance liquid chromatography with diode array detection,ultra high performance liquid chromatography with tandem quadrupole mass spectrometry,and chemometrics

Plants of Erycibe are widely used in traditional Chinese medicine for the treatment of joint pain and rheumatoid arthritis. With the reduction of Erycibe resources in the wild, Porana sinensis has been widely used as a substitute. However, it is important to understand the chemical distinctions between the two kinds of plants and identify their individual chemical markers. In this study, multiwavelength chromatographic fingerprint and precursor ion fingerprint techniques were used in conjunction with chemometric tools to fingerprint and thus differentiate between plant samples. The similar results obtained from different fingerprints prove the reliability of the two fingerprints. Results obtained from principal component analysis and orthogonal projection to latent structures discriminant analysis identified similarities between the chemical components of P. sinensis and plants of Erycibe. However, concentrations of 4‐caffeoylquinic acid, 3,5‐dicaffeoylquinic acid, 3,4‐dicaffeoylquinic acid, and 4,5‐dicaffeoylquinic acid were higher in P. sinensis than in plants of Erycibe, suggesting that P. sinensis may be more effective in medical treatments of some diseases than Erycibe.     

Chemical fingerprint of Ganmaoling granule by double‐wavelength ultra high performance liquid chromatography and ultra high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry

A method incorporating double‐wavelength ultra high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry was developed for the investigation of the chemical fingerprint of Ganmaoling granule. The chromatographic separations were performed on an ACQUITY UPLC HSS C₁₈ column (2.1 × 50 mm, 1.8 μm) at 30°C using gradient elution with water/formic acid (1%) and acetonitrile at a flow rate of 0.4 mL/min. A total of 11 chemical constituents of Ganmaoling granule were identified from their molecular weight, UV spectra, tandem mass spectrometry data, and retention behavior by comparing the results with those of the reference standards or literature. And 25 peaks were selected as the common peaks for fingerprint analysis to evaluate the similarities among 25 batches of Ganmaoling granule. The results of principal component analysis and orthogonal projection to latent structures discriminant analysis showed that the important chemical markers that could distinguish the different batches were revealed as 4,5‐di‐O‐caffeoylquinic acid, 3,5‐di‐O‐caffeoylquinic acid, and 4‐O‐caffeoylquinic acid. This is the first report of the ultra high performance liquid chromatography chemical fingerprint and component identification of Ganmaoling granule, which could lay a foundation for further studies of Ganmaoling granule.     

Characterization of the multiple chemical components of Glechomae Herba using ultra high performance liquid chromatography coupled to quadrupole‐time‐of‐flight tandem mass spectrometry with diagnostic ion filtering strategy

Glechomae Herba is a traditional Chinese medicine used for the treatment of urolithiasis, cholelithiasis, and urinary tract infections in China. Identification of chemical constituents is helpful to discover the potential active ingredients. However, this significant work is stymied by complex chemical constituents. Therefore, an ultra high performance liquid chromatography coupled to quadrupole‐time‐of‐flight tandem mass spectrometry analysis with diagnostic product ions and neutral loss filtering strategy was established for chemical profiling of Glechomae Herba. The diagnostic product ions and neutral loss filtering strategy simplified spectral elucidation. A total of 120 compounds, including 10 chlorogenic acids, 10 gallic acids, 21 phenylpropionic acids, and 77 flavonoids, were reasonably identified in Glechomae Herba. Sixty‐five constituents were first discovered in Glechomae Herba. Four types of chlorogenic acids (caffeoylquinic acid, feruloylquinic acid, p‐coumaroylquinic acid, and di‐caffeoylquinic acid), three types of galloylglucoses (di‐O‐galloyl‐glucose, tri‐O‐galloyl‐glucose, and tetra‐O‐galloyl‐glucose), three types of phenylpropionic acid skeletons (p‐coumaric acid, caffeic acid, and rosmarinic acid) and five types of flavonoid aglycone skeletons (apigenin, kaempferol, luteolin, quercetin, and chrysin) were identified in Glechomae Herba. The results indicated that the developed strategy was feasible and rational technique for identifying the complex chemical constituents in Glechomae Herba.     

Liquid chromatography coupled with ultraviolet absorbance detection,electrospray ionization,collision‐induced dissociation and tandem mass spectrometry on a triple quadrupole for the on‐line characterization of polyphenols and methylxanthines in green coffee beans

Liquid chromatography coupled with a photodiode array detector, electrospray ionization, collision‐induced dissociation and tandem mass spectrometry (LC‐DAD/ESI‐CID‐MS/MS) on a triple quadrupole (QqQ) has been used to detect and characterize polyphenols and methylxanthines in green coffee beans: three phenolic acids (caffeic acid, ferulic acid and dimethoxycinnamic acid), three isomeric caffeoylquinic acids (M_r 354), three feruloylquinic acids (M_r 368), one p‐coumaroylquinic acid (M_r 338), three dicaffeoylquinic acids (M_r 516), three feruloyl‐caffeoylquinic acids (M_r 530), four p‐coumaroyl‐caffeoylquinic acids (M_r 500), three diferuloylquinic acids (M_r 544), six dimethoxycinnamoyl‐caffeoylquinic acids (M_r 544), three dimethoxycinnamoyl‐feruloylquinic acids (M_r 558), six cinnamoyl‐amino acid conjugates, three cinnamoyl glycosides, and three methylxanthines (caffeine, theobromine and theophylline). Dimethoxycinnamic acid, three isomers of dimethoxycinnamoyl‐caffeoylquinic acids and another three of dimethoxycinnamoyl‐feruloylquinic acids, as well as the three cinnamoyl glycosides, had not previously been reported in coffee beans. Structures have been assigned on the basis of the complementary information obtained from UV‐visible spectra, relative hydrophobicity, scan mode MS spectra, and fragmentation patterns in MS² spectra (both in the positive and negative ion modes) obtained using a QqQ at different collision energies. A structure diagnosis scheme is provided for the identification of different isomers of polyphenols and methylxanthines. Copyright © 2009 John Wiley & Sons, Ltd.     

Determination of asperosaponin VI in dog plasma by high-performance liquid chromatography-tandem mass spectrometry and its application to a pilot pharmacokinetic study

A sensitive and rapid liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method has been developed and validated for the determination of asperosaponin VI in beagle dog plasma using glycyrrhizic acid as the internal standard (IS). Plasma samples were simply pretreated with methanol for deproteinization. Chromatographic separation was performed on a Hedera ODS‐2 column using mobile phase of methanol–10 mm ammonium acetate buffer solution containing 0.05% acetic acid (71:29, v/v) at a flow rate of 0.38 mL/min. Asperosaponin VI and the IS were eluted at 2.8 and 1.9 min, respectively, ionized in negative ion mode, and then detected by multiple reaction monitoring. The detection used the transitions of the deprotonated molecules at m/z 927.5 → 603.4 for asperosaponin VI and m/z 821.4 → 645.4 for glycyrrhizic acid (IS). The assay was linear over the concentration range of 0.15–700 ng/mL and was successfully applied to a pilot pharmacokinetic study in beagle dogs. Copyright © 2011 John Wiley & Sons, Ltd.     

A UFLC–MS/MS method for the simultaneous determination of eight bioactive constituents from red wine and dealcoholized red wine in rat plasma: Application to a comparative pharmacokinetic study

To explore whether alcohol has an effect on the pharmacokinetic behavior of phenolic acids, the main bioactive constituents in red wine, a highly sensitive and simple ultra‐fast liquid chromatography coupled with triple quadrupole mass spectrometry (UFLC–MS/MS) method was developed for simultaneous quantitation of eight phenolic acids in plasma samples. Plasma samples were extracted by liquid–liquid extraction and the chromatographic separation was achieved on a Zorbax SB‐C₁₈ column within 7.0 min. Results of the validated method revealed that all of the calibration curves displayed good linear regression (r > 0.99). The intra‐ and inter‐day precisions of the analytes were <14.0% and accuracies ranged from ?8.5 to 7.3%. The extraction recoveries of the analytes were from 71.2 to 110.2% and the matrix effects ranged from 86.2 to 105.5%. The stability of these compounds under various conditions satisfied the requirements of biological sample measurement. The method was successfully applied to a comparative pharmacokinetic study of phenolic acids in rat plasma. For gallic acid and gentisic acid, the parameters AUC_0–t and AUC_0–∞ increased remarkably (p < 0.05) after oral administration of red wine, which suggested that alcohol might enhance their absorption. This is the first report to compare the pharmacokinetic behavior of phenolic acids in red wine and dealcoholized red wine.     

Simultaneous determination of ten flavonoids of crude and wine‐processed Radix Scutellariae aqueous extracts in rat plasma by UPLC‐ESI‐MS/MS and its application to a comparative pharmacokinetic study

Radix Scutellariae (RS) is a herbal medicine with various pharmacological activities to treat inflammation, respiratory and gastrointestinal infections, etc. In this study, a rapid, sensitive and selective UPLC‐ESI‐MS/MS method was developed for simultaneous determination of 10 flavonoids – scutellarin, scutellarein, chrysin, wogonin, baicalein, apigenin, wogonoside, oroxylin A‐7‐O‐glucuronide, oroxylin A and baicalin – from RS aqueous extracts in rat plasma with propyl paraben as internal standard (IS). Chromatographic separation was achieved on a C₁₈ column using gradient elution with the mobile phase consisting of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. The detection was performed in multiple reaction monitoring mode using electrospray ionization in negative mode. The validated method showed good linearity over a wide concentration range (r >0.9935). The intra‐ and interday assay variabilities were <9.5% and <12.4% for all analytes, respectively. The extraction recovery ranged from 71.2 to 89.7% for each analyte and IS. This method was successfully applied to pharmacokinetic comparision after oral administration of crude and wine‐processed RS aqueous extracts. There were significant differences in some pharmacokinetic parameters of most analytes between crude and wine‐processed RS. This suggested that wine‐processing exerted effects absorption of most flavonoids. Copyright © 2014 John Wiley & Sons, Ltd.     

Rapid preparative separation of six bioactive compounds from Gentiana crassicaulis Duthie ex Burk. using microwave‐assisted extraction coupled with high‐speed counter‐current chromatography

A rapid method combining microwave‐assisted extraction (MAE) and high‐speed counter‐current chromatography (HSCCC) was applied for preparative separation of six bioactive compounds including loganic acid ( I ), isoorientin‐4′‐O‐glucoside ( II ), 6′‐O‐β‐d ‐glucopyranosyl gentiopicroside ( III ), swertiamarin ( IV ), gentiopicroside ( V ), sweroside ( VI ) from traditional Tibetan medicine Gentiana crassicaulis Duthie ex Burk. MAE parameters were predicted by central composite design response surface methodology. That is, 5.0 g dried roots of G. crassicaulis were extracted with 50 mL 57.5% aqueous ethanol under 630 W for 3.39 min. The extract (gentian total glycosides) was separated by HSCCC with n‐butanol/ethyl acetate/methanol/1% acetic acid water (7.5:0.5:0.5:3.5, v/v/v/v) using upper phase mobile in tail‐to‐head elution mode. 16.3, 8.8, 12., 25.1, 40.7, and 21.8 mg of compounds I–VI were obtained with high purities in one run from 500 mg of original sample. The purities and identities of separated components were confirmed using HPLC with photo diode array detection and quadrupole TOF‐MS and NMR spectroscopy. The study reveals that response surface methodology is convenient and highly predictive for optimizing extraction process, MAE coupled with HSCCC could be an expeditious method for extraction and separation of phytochemicals from ethnomedicine.     

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 six index constituents and comparative analysis of four ethnomedicines from genus Gentiana using a UPLC‐UV‐MS method

Many species from genus Gentiana (Gentianaceae) have a long history of applications as folk medicines in the world. A simple rapid UPLC‐UV‐MS/MS method has been developed and validated for the simultaneous determination of six index constituents (gentiopicroside, swertiamarin, loganic acid, sweroside, mangiferin and ferulic acid) from the four ethnomedicines (G. rigescens Franch. ex Hemsl., G. rhodantha Franch. ex Hemsl., G. scabra Bunge and G. farreri Balf. f.). The UPLC analysis was performed on Shim‐Pack XR‐ODS III (150 × 2.0 mm, 2.2 µm). The mobile phase was consisted of acetonitrile–0.1% formic acid water using gradient elution. The wavelength 242 nm was chose for the four iridoids as well as mangiferin and 320 nm was set for ferulic acid. Mass spectrometry was applied for identification and quantification for analytes with low concentration. All the regression equations revealed a good linear relationship (R² > 0.9993). Accuracy and precision were all within the required limits. The chromatogram fingerprints analysis combined with principal component analysis showed the similarity values of the four species were <0.788 while the similarity values of G. scabra Bunge and G. rigescens Franch. ex Hemsl. were >0.993, which provided reasonable foundation for utilization and clinical application of the four ethnomedicines. This developed method appears to be a useful tool for quality control of the four ethnomedicines. Copyright © 2014 John Wiley & Sons, Ltd.