3beta-(2-O-alpha-L-Rhamnopyranosyl-beta-D-xylopyranosyloxy)-23-hydroxyoleana-12-ene-28-oic acid 6-O-beta-D-glucopyranosyl-beta-D-glucopyranosyl ester

UHPLC-MS/MS quantification combined with chemometrics for the comparative analysis of different batches of raw and wine-processed Dipsacus asper.[Pubmed:28218445]

J Sep Sci. 2017 Apr;40(8):1686-1693.

A rapid and sensitive ultra-high performance liquid chromatography with tandem mass spectrometry approach was established for the simultaneous determination of 4-caffeoylquinic acid, loganic acid, chlorogenic acid, loganin, 3,5-dicaffeoylquinic acid, dipsacoside B, asperosaponin VI, and sweroside in raw and wine-processed Dipsacus asper. Chloramphenicol and glycyrrhetinic acid were employed as internal standards. The proposed approach was fully validated in terms of linearity, sensitivity, precision, repeatability as well as recovery. Intra- and interassay variability for all analytes were 2.8-4.9 and 1.7-4.8%, respectively. The standard addition method determined recovery rates for each analytes (96.8-104.6%). In addition, the developed approach was applied to 20 batches of raw and wine-processed samples of Dipsacus asper. Principle component analysis and partial least squares-discriminate analysis revealed a clear separation between the raw group and wine-processed group. After wine-processing, the contents of loganic acid, chlorogenic acid, dipsacoside B, and asperosaponin VI were upregulated, while the contents of 3,5-dicaffeoylquinic acid, 4-caffeoylquinic acid, loganin, and sweroside were downregulated. Our results demonstrated that ultra-high performance liquid chromatography with tandem mass spectrometry quantification combined with chemometrics is a viable method for quality evaluation of the raw Dipsacus asper and its wine-processed products.

Comparative pharmacokinetic analysis of extracts of crude and wine-processed Dipsacus asper in rats by a sensitive ultra performance liquid chromatography-tandem mass spectrometry approach.[Pubmed:27710888]

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Nov 15;1036-1037:33-41.

The purpose of this study is to establish and validate an UPLC-MS/MS approach to determine 4-caffeoylquinic acid, chlorogenic acid, 3,5-dicaffeoylquinic acid, loganic acid, loganin, sweroside, dipsacoside B and asperosaponin VI from extracts of crude and wine-processed Dipsacus asper in biological samples and apply the approach to a comparative pharmacokinetic study. A Waters BEH C18 UPLC column was employed with acetonitrile/0.2% formic acid-water as mobile phases. The mass analysis was carried out in a triple quadrupole mass spectrometer using multiple reaction monitoring (MRM) with negative scan mode. A one-step protein precipitation by acetonitrile was performed to extract the eight analytes from plasma. Our results revealed that all of the calibration curves displayed good linear regression (r(2)>0.9990). The lower limits of quantification (LLOQ) were determined as 10.0, 9.6, 8.9, 9.1, 9.2, 9.8, 10.1 and 9.8ng/mL. The intra-day and inter-day precisions (RSD) of the eight compounds at high, medium and low levels were less than 4.94% and the bias of the accuracies ranged from -3.89% to 3.95%.The extraction recoveries of the eight compounds were from 90.4% to 100.2% and the matrix effects ranged from 89.3% to 100.1%. The stabilities of these compounds were investigated by analyzing six replicates of QC samples at three different concentrations following storage at 25 degrees C for 4h, -80 degrees C for 30days, three-freeze-thaw cycles, and 4 degrees C for 24h. All the samples showed satisfactory precision and accuracy after various stability tests. Pharmacokinetic parameters were estimated using a non-compartment model. Compared with the crude group, the parameters of Cmax and AUC0-t of 4-caffeoylquinic acid, loganic acid, loganin and asperosaponin VI increased remarkably (p<0.05) after oral administration of the aqueous extract of wine-processed Dipsacus asper, indicating that wine-processing could enhance bioavailability of 4-caffeoylquinic acid, loganic acid, loganin and asperosaponin VI.

Simultaneous determination of five triterpenoid saponins in different parts of Lonicera macranthoides by RRLC-MS/MS method.[Pubmed:27455548]

Pharmazie. 2016 Jun;71(6):306-10.

A rapid resolution liquid chromatography-tandem mass spectrometry (RRLC-MS/MS) method was developed and validated for the determination of five major saponins (macranthoidin B, macranthoidin A, dipsacoside B, akebiasaponin D, and dipsacoside A) in the flower bud, stem, and leaf parts of Lonicera macranthoides. Chromatographic separation was performed on a ZORBAX SB-C18 column (2.1 x 50 mm, 1.8 mum). Acetonitrile and 0.1% aqueous formic acid were adopted as mobile phase. Detection was carried out on a triple quadrupole mass spectrometer in the negative ion mode using an electrospray source. Multiple reaction monitoring (MRM) mode was employed. The established method showed good linearity (r2 >/= 0.9994) for all the analytes within the test ranges and the recoveries were 95.19-103.28%. Desirable intra-day and inter-day precision as well as repeatability were obtained with relative standard deviations (RSDs) less than 5%. The method was simple, sensitive, accurate and performed well in application to the sample determination within a short analysis time of 15 min. The saponin profiles of different parts of Lonicera macranthoides were obtained based on the quantitative data, showing that the flower bud contained much higher level of saponins than the stem and leaf by several orders of magnitude, and that the quantity ratios varied remarkable between these three part. The conclusions might provide scientific evidences for the reasonable application of Lonicera macranthoides, and the proposed RRLC-MS/MS method might be useful for the quality control of this medicinal plant.

Rapid quantitative analysis of adulterant Lonicera species in preparations of Lonicerae Japonicae Flos.[Pubmed:26420337]

J Sep Sci. 2015 Dec;38(23):4014-20.

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.

[Effects of Aphid Occurring on Lonicera macranthoides Bud Yield and Quality].[Pubmed:26214864]

Zhong Yao Cai. 2015 Jan;38(1):8-10.

OBJECTIVE: To study the effects of aphid occurring on the quality of Lonicera macranthoides bud. METHODS: The fresh and dry bud yields of different aphid damage in the same plant were caculated. Contents of chlorogenic acid caffeic acid, luteolin-7-O-gluco-side, isochlbrogenic acid A, macranthoidin B and dipsacoside B in Lonicera macranthoides bud damaged by aphid were determined by HPLC-ELSD. RESULTS: Aphid significantly affected the contents of luteolin-7-O-glucoside, macranthoidin B, dipsacoside B, and chlorogenic acid (P < 0. 01 or P <0. 05), but had no obvious effect on the content of caffeic acid and isochlorogenic acid A in the bud of Lonicera macranthoides. CONCLUSIONS: It is supposed to regulate prevention and control of aphids in Lonicera macranthoides GAP base.

[Research on quality standards of zhuang medicine Lonicerae dasystylae flos].[Pubmed:23724691]

Zhongguo Zhong Yao Za Zhi. 2013 Mar;38(5):762-7.

OBJECTIVE: To establish quality standard of Zhuang medicine Lonicera dasystyla, and provide scientific basis for the quality control of L. dasystyla. METHOD: Characteristics of materia medica, microscopic features, TLC indentification, inspection, extractum and determination of chlorogenic acid, macranthoidin B, dipsacoside B were carried out through the experience, microscopic, physical and chemical methods, respectively. The standard of quality control was formulated thereafter. RESULT: The characteristics of materia medica, microscopic features, TLC indentification were specified, the average contents of water, total ash, acid-insoluble ash, alcohol-soluble extracts, chlorogenic acid were 11.6%, 6.6%, 0.2% , 24.4%, 1.16%, respectively, the total amount of macranthoidin B and dipsacoside B was 3.13%. Quality standard of L. dasystyla was proposed according to experimental results. CONCLUSION: The quality of L. dasystyla can be controlled effectively with the quality standard.

[Chemical constitutents from pre-formulation of lonicerae japonicae flos in shuanghuanglian lyophilized powder for injection].[Pubmed:22007545]

Zhongguo Zhong Yao Za Zhi. 2011 Jun;36(12):1613-9.

OBJECTIVE: To research the chemical constitutents for the pre-formulation of Lonicerae Japonicae Flos (the dried buds of Lonicera japonica) in Shuanghuanglian lyophilized powder for injection and provide substance foundation for the adverse reaction of Shuanghuanglian lyophilized powder for injection. METHOD: The chemical constituents were isolated by column chromatography and preparative HPLC. All structures were characterized by the spectroscopic methods including ESI-MS, 1H-NMR, 13C-NMR, and compared with data in the literature. RESULT: Twenty compounds were isolated and identified as sophoraricoside(1), luteolin-7-O-beta-D-glucopyranoside(2), rutin(3), quercetin(4), 3,5-O-dicaffeoyl quinic acid methyl ester(5), 4,5-O-dicaffeoyl quinic acid methyl ester(6), 3,4-O-dicaffeoyl quinic acid methyl ester(7), 4,5-dicaffeoyl quinic acid(8), 3,4-dicaffeoyl quinic acid(9), chlorogenic acid(10), epi-vogeloside (11), sweroside(12), vogeloside(13), secoxyloganin(14), macranthoidin A(15), macranthoidin B(16), loniceroside A(17), loniceroside B(18), loniceroside C(19), dipsacoside B(20). CONCLUSION: Compound 1 was identified in genus Lonicera for the first time and compounds 1-20 were isolated from the pre-formulation for the first time.

Biologically active triterpenoids from Cephalaria ambrosioides.[Pubmed:19152225]

Planta Med. 2009 Feb;75(2):163-7.

The roots of Cephalaria ambrosioides yielded a new triterpene, 6alpha-hydroxyhederagenic acid ( 1), in addition to the known triterpene hederagenic acid ( 2) and four corresponding saponins, leontoside A (or akeboside Stb) ( 3), kalopanax saponin A (or alpha-hederin) ( 4), saponin PG (or sapindoside B) ( 5), and dipsacoside B ( 6). Their structures have been elucidated on the basis of their spectral data (MS, 1 D and 2 D NMR) and by some chemical transformations. The extract and all isolated compounds were evaluated for their antimicrobial, molluscicidal and IN VITRO cytotoxic activities. All compounds showed strong antimicrobial activity (MIC values 1.80 - 2.50 microg/mL), with 5 and 6 exhibiting activities comparable to those of standard antibiotics. Moreover, compounds 3 - 5 were active against all assayed cancer cell lines, whereas compounds 3 and 4 exhibited higher activities against Biomphalaria Glabrata snails, with minimum inhibitory concentrations of 5.4 and 6.2 microg/mL, respectively.

Liquid chromatography-mass spectrometry analysis of macranthoidin B, macranthoidin A, dipsacoside B, and macranthoside B in rat plasma for the pharmacokinetic investigation.[Pubmed:19097951]

J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Jan 15;877(3):159-65.

A liquid chromatography-electrospray ionization-mass spectrometry method has been developed and validated for identification and quantification of four major bioactive saponins in rat plasma after oral administration of extraction of saponins from Flos Lonicerae, i.e., macranthoidin B, macranthoidin A, dipsacoside B, and macranthoside B. Plasma samples were extracted with solid-phase extraction, separated on a Shim-pack CLC-ODS column and detected by MS in negative selective ion monitoring mode. Calibration curves offered linear ranges of two orders of magnitude with r(2)>0.999. The method showed the low limit quantification of 7.72, 6.06, 7.16, and 1.43 ng/mL for macranthoidin B, macranthoidin A, dipsacoside B, and macranthoside B, respectively. The inter- and intra-CV precision (R.S.D.) were all within 10% and accuracy (% bias) ranged from -10 to 10%. The overall recovery was more than 70%. This developed method was subsequently successfully applied to pharmacokinetic profiles of the four saponins in rats. After oral administration of extraction of saponins in rats, the concentration-time course was found to be the double peaks of curve.

Quality evaluation of Flos lonicerae through a simultaneous determination of seven saponins by HPLC with ELSD.[Pubmed:15861786]

J Chromatogr A. 2005 Apr 8;1070(1-2):43-8.

A new HPLC coupled with evaporative light scattering detection (ELSD) method has been developed for the simultaneous quantitative determination of seven major saponins, namely macranthoidin B (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-beta-D-glucopyranosyl(6-->1)-O-beta-D-glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-alpha-L-arabinopyranosyl(2-->1)-O-alpha-L-rhamnopyranoside (7) in Flos Lonicerae, a commonly used traditional Chinese medicine (TCM) herb. Simultaneous separation of these seven saponins was achieved on a C18 analytical column. The mobile phase consisted of (A) acetonitrile-acetic acid (95:0.5) and (B) 0.5% aqueous acetic acid using a gradient elution of 29%A at 0-10 min, 29-46%A at 10-25 min and 46%A at 25-30 min. The drift tube temperature of ELSD was set at 106 degrees C, and with the nitrogen flow-rate of 2.6 l/min. All calibration curves showed good linear regression (r2>0.9922) within test ranges. This method showed good reproducibility for the quantification of these seven saponins in Flos Lonicerae with intra- and inter-day variations of less than 3.0% and 6.0%, respectively. The validated method was successfully applied to quantify seven saponins in five sources of Flos Lonicerae, which provides a new basis of overall assessment on quality of Flos Lonicerae.

[Triterpene saponins from Pulsatilla cernua].[Pubmed:11372442]

Yao Xue Xue Bao. 2000 Oct;35(10):756-9.

AIM: To investigate the chemical constituents of the roots of Pulsatilla cernua (Thunb.) Bercht. et Opiz. METHODS: Column chromatography (including D101 macroporous resin, silica gel and ODS) and HPLC were used to separate the chemical constituents whose structures were elucidated by FAB-MS, NMR (1D and 2D) and hydrolysis methods. RESULTS: Six compounds were isolated and established as hederagenin 3-O-alpha-L-rhamnopyranosyl (1-->2)-alpha-L-arabinopyranoside (pulsatilla saponin A, 1), hederagenin 3-O-beta-D-glucopyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1-->2)-alpha-L- arabinopyranoside (2), hederagenin' 3-O-alpha-L-rhamnopyranosyl(1-->2)[beta-D- glucopyranosyl(1-->4)]-alpha-L-arabinopyranoside (pulsatilla saponin D, 3), 3-O-alpha-L-rhamnopyranosyl (1-->2)-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranosyl ester (dipsacoside B, 4), 3-O-alpha-L-rhamnopyranosyl(1-->2) [beta-D-glucopyranosyl(1-->4)]-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl ester (cernuoside C, 5) and daucosterol (6), respectively. CONCLUSION: Compound 5 is a new compound and named as cernuoside C. Compounds 2, 4 and 6 were isolated from Pulsatilla cernua for the first time.

[Studies on the chemical constituents of Lonicera macranthoides Hand. -Mazz].[Pubmed:8213170]

Yao Xue Xue Bao. 1993;28(4):273-81.

Three bisdesmosidic triterpenoid saponins I-III, have been isolated from the flowers of Lonicera macranthoides Hand. -Mazz. By spectral (IR, MS, 1H-NMR and 13C-NMR) and chemical methods, I was proved to be identical with the known dipsacoside B, II and III were new compounds named macranthoidin A and macranthoidin B. Their structures were identified as follows: The prosapogenin of II, macranthoside A(IIb), was elucidated as 3-O-beta-D-glucopyranosyl-(1-3)-alpha-L-rhamnopyranosyl-(1-2)-alpha-L- arabinopyranosyl 3 beta, 23-dihydroxyl-olean-12-en-28-olic acid, II was established as 3-O-beta-D-glucopyranosyl-(1-3)-alpha-L-rhamnopyranosyl-(1-2)-alpha-L- arabinopyranosyl 3 beta,23-dihydroxyl-olean-12-en-28-O-beta-D-glucopyranosyl-(1-6)-beta-D- glucopyranoside. The prosapogenin of III, macranthoside B(IIIb), was elucidated as 3-O-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-3)-alpha-L- rhamnopyranosyl-(1-2)-alpha-L-arabinopyranosyl 3 beta,23-dihydroxyl-olean-12-en-28-olic acid, III was established as 3-O-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-3)-alpha-L- rhamnopyranosyl-(1-2)-alpha-L-arabinopyranosyl 3 beta,23-dihydroxyl-olean-12-en-28-O-beta-D-glucopyranosyl-(1-6)-beta-D- glucopyranoside. IIb and IIIb have not been reported in the literature.