Effects of isolated scenting on the taste quality of broken green tea based on metabolomics.

Liquid chromatography-mass spectrometry (LC-MS) combined with multivariate analysis were used to characterize the nonvolatile compounds of broken green tea and explore the effect of isolated scenting on metabolic profile and taste quality of broken green tea in this research. A total of 236 nonvolatile compounds were identified and 13 compounds were believed to be the key characteristic taste compounds of scented broken green tea. Meanwhile, the optimal isolated scenting time of broken green tea was determined to be 10 h based on the sensory evaluation and PLS results. The contents and types of flavonoids, organic acids and catechins lead to the difference of taste quality at different scenting times. Overall, these findings provided a theoretical basis for scenting to improve the taste of broken green tea, and provide a new idea for improving the taste of broken green tea.

[Main components in different varieties of Xihuangcao by UPLC-Q-TOF-MS and UPLC-DAD].

This study established an ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) method to analyze the main components in different varieties of Xihuangcao and established a UPLC-DAD method to simultaneously determine the five active components(caffeic acid, rosmarinic acid, schaftoside, isoschaftoside, and oridonin).The chromatographic separation was performed on a Waters ACQUITY UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 µm) with a gradient elution of methanol(B)-water containing 0.1% formic acid(A) at a flow rate of 0.3 mL·min~(-1).The column temperature was 30 ℃.The Q-TOF-MS discriminant analysis was performed under positive electrospray ion mode and the split ratio was 1∶1. Quantitative analysis was carried out by UPLC-DAD.The determination wavelength was set at 245 nm.Thirty-two main components of Xihuangcao were separated and identified by UPLC-Q-TOF-MS, where 19 were identified in Rabdosia serra, nine in R.nervosa, 10 in R.lophanthoides, 15 in R.lophanthoides var.graciliflora, 10 in R.lophanthoides var.gerardianus, and seven in R.stracheyi.The UPLC-DVD method was developed for simultaneously determining five active components in different varieties of Xihuangcao.The standard curves for five compounds showed good linearity with correlation coefficients higher than 0.999 0.The precision, repeatability, and stability were good.The average recoveries(n=6) were between 97.01% and 102.7% with RSD<3.0%.The results of UPLC-Q-TOF-MS analysis provided a scientific basis for the use of R.stracheyi as a medicinal material of Xihuangcao and the equivalent use of R.lophanthoides var.gerardianus with R.lophanthoides var.graciliflora to some extent.The UPLC-DAD method for simultaneously determining five active components is simple, rapid, and accurate.This study can provide the basis for the quality control of different varieties of Xihuangcao.

Flavonoid pattern inheritance in the allopolyploid Spartina anglica - Comparison with the parental species S. maritima and S. alterniflora.

The invasive species Spartina anglica arose in Europe by a cross between the Afro-European species S. maritima (native, paternal ancestor) and the introduced North American S. alterniflora (invasive, maternal ancestor). Aqueous methanolic extracts were prepared from plant tissue for chemotaxonomical comparison between the three species and determination of the phenolic pattern inheritance in S. anglica. A total of 20 phenolic compounds were detected in the aerial tissues of S. anglica and S. alterniflora, but only seven in S. maritima. They were isolated from their respective crude extracts, and their structures were determined according to spectroscopic data analysis and chemical evidence. They all belong to the flavonoid class, with 13 of them identified as C-glycoflavonoid and seven as O-glycoflavonoid. All these products were detected for the first time from S. anglica, fourteen of them for the first time from S. alterniflora, and three of them for the first time from S. maritima. The individual concentrations in the three species were determined by quantitative HPLC. The two parental species were found to differ markedly in their foliar phenolic fingerprint, whereas that of S. anglica showed a clear maternal dominance. Eight of the fourteen major compounds identified were of maternal origin among which, six were over-expressed, only three were from paternal origin but under-regulated, while two originated from the two parents. As far as we know, this work represents the first exhaustive report of the phenolic fingerprints of S. alterniflora and S. anglica and of the phenolic pattern inheritance in S. anglica. The similarity in the phenolic chemistry of the introduced and invasive S. alterniflora to its progeny could play a role in the physiological vigour and invasion success of S. anglica. This work provide a foundation for further studies, considering the reported biological activities of C-glycosidic flavonoids and tricin derivatives, and the lack of knowledge of the ecological chemistry of the genus Spartina.

Antioxidant activity and polyphenol composition of sugarcane molasses extract.

The antioxidant activity of sugarcane molasses ethanol extract (ME) and its fraction (ME-RBF) was evaluated using ABTS, ORAC 6.0 and CAA assays and ME-RBF demonstrated 26-fold, 12-fold and 2-fold higher values, respectively than ME. Likewise, total polyphenol and flavonoid concentration in ME-RBF are more than 10-fold higher than ME, that suggested antioxidant activity is correlated with polyphenol composition. Quantitative analysis of 13 polyphenols (chlorogenic acid, caffeic acid, sinapic acid, syringic acid, vanillin, homoorientin, orientin, vitexin, swertisin, diosmin, apigenin, tricin and diosmetin) was carried out by LCMS. MS/MS analysis allowed the tentative identification of seven apigenin-C-glycosides, three methoxyluteolin-C-glycosides and three tricin-O-glycosides some of which have not been reported in sugarcane before to the best of our knowledge. The results demonstrated that sugarcane molasses can be used as potential source of polyphenols that can be beneficial to health.

Identification and quantitative analysis of phenolic glycosides with antioxidant activity in methanolic extract of Dendrobium catenatum flowers and selection of quality control herb-markers.

Recently, the flowers of Dendrobium catenatum (D. officinale) have been approved as new food ingredient. This study aimed to investigate the herb-markers and their antioxidant activities in methanolic extracts of D. catenatum flowers, and to establish the quality evaluation methods for raw materials and their products of flower by HPLC. The methanolic extract of 11 strains of D. catenatum flowers were found to contain a high content of total phenol and flavonoids, and they possessed potential antioxidant capacities based on DPPH radical scavenging assay. A total of 21 phenolic herb-markers were selected according to the similarity and principal component analysis of the chromatographic fingerprinting profiles. Their structures were further elucidated by UV, HPLC-DAD-ESI-QTOF-MS/MS and NMR analyses. The identified compounds included 2 phenylpropanoids, 11C-glycosylflavones and 6 O-glycosylflavones, which could be employed as the indicators for quantitative evaluation of the quality and authenticity of the flowers. Based on the pre-column DPPH/ABTS+-HPLC analysis, the major compounds contributed to the antioxidative activity were identified as 1-O-caffeoyl-ß-D-glucoside, rutin and isoquercitrin, all of which, were also the most abundant constituents in the methanolic extract. The results suggest the potential of D. catenatum flowers as a new antioxidant resources for medicinal and food products.

Pharmacokinetics and absolute bioavailability of isoschaftoside in rat by LC-MS/MS / 中国药科大学学报

@#The aim of this study was to develop a highly sensitive and specific LC-MS/MS method to explore the pharmacokinetic properties and absolute bioavailability of isoschaftoside in rats. Blood sampling was performed at different time points after intragastric administration of isoschaftoside(1. 5, 3. 0, 6. 0 mg/kg)and 0. 5 mg/kg by intravenous injection. Isoschaftoside was analyzed by a validated LC-MS/MS method in plasma; the pharmacokinetic parameters and absolute bioavailability were evaluated by software DAS 3. 0. The results showed that the linear concentration ranges of isoschaftoside was 1. 0- 500. 0 ng/mL(r=0. 997 6). The precision, accuracy, matrix effect, sensitivity, dilution reliability and stability met the requirements of biological sample analysis. For ig administration of isoschaftoside(1. 5, 3. 0, 6. 0 mg/kg), the pharmacokinetic parameter cmax was(109. 34±22. 87), (259. 84±95. 35)and(499. 26±288. 09)ng/mL; AUC0-t was(310. 57±46. 18), (552. 67±207. 14)and(1 075. 03±371. 19)h ·ng/mL; t1/2 was(2. 36±0. 22), (2. 91±0. 19)and(3. 04±0. 86)h; tmax was(1. 03±0. 25), (1. 18±0. 17)and(1. 5±0. 43)h; MRT0-t was(11. 33±1. 53), (11. 27±1. 09)and(8. 29±0. 76)h, respectively. For iv administration of isoschaftoside(0. 5 mg/kg), the pharmacokinetic parameter AUC0-t was(1 536±421. 3)h ·ng/mL; t1/2 was(2. 57±0. 46)h; MRT0-t was(9. 55±2. 37)h. Furthermore, the absolute bioavailability was 6. 73%, 5. 99%, 5. 80%, respectively. The LC-MS/MS analysis method established in this study was accurate and sensitive, so it can be applied to the pharmacokinetic study of isoschaftoside.

Costus spiralis (Jacq.) Roscoe: A Novel Source of Flavones with α-Glycosidase Inhibitory Activity.

Costus spiralis, a plant used in traditional Brazilian medicine for the treatment of complications in diabetes, was investigated. Assay of hexane, ethyl acetate, methanol, and aqueous fractions obtained by partition of a crude methanol extract of dried leaves of C. spiralis revealed that AGI activity was confined to the ethyl acetate fraction. Purification of this fraction yielded schaftoside and isoschaftoside. The AGI activities of the two flavones were lower than, but comparable with, that of the anti-diabetic drug acarbose. In contrast, the IC50 value of the ethyl acetate fraction was 1.95-, 2.34-, and 2.22-fold higher than those of acarbose, schaftoside, and isoschaftoside, respectively. The results demonstrate for the first time that schaftoside and isoschaftoside are responsible, in part, for the AGI activity of C. spiralis. Our study suggests that further investigations into C. spiralis may lead to the discovery of additional compounds with antihyperglycemic activity.

Simultaneous Determination of Caffeic Acid,Isoschaftoside and Schaftoside in Xiaoyan Lidan Tablet by RP-HPLC / 中国药房

ABSTARCT OBJECTIVE:To establish a method for the simultaneous determination of caffeic acid,isoschaftoside and schafto-side in Xiaoyan lidan tablet. METHODS:RP-HPLC was performed on the column of Phenomenex C18 with mobile phase of metha-nol- 0.1% phsphate acid(gradient elution)at flow rate of 0.8 ml/min,the detection wavelength was 334 nm,column temperature was 25 ℃,and the injection volume was 10 μl. RESULTS:The linear range was 0.10-2.81 μg for caffeic acid(r=0.999 9), 0.20-5.63 μg for isoschaftoside(r=0.999 9)and 0.10-2.63 μg for schaftoside(r=0.999 9);RSDs of precision,stability and repro-ducibility tests were lower than 3%;recoveries were 96.3%-100.4%(RSD=1.59%,n=9),97.3%-101.2%(RSD=1.28%,n=9) and 96.6%-100.6%(RSD=1.39%,n=9),respectively. CONCLUSIONS:The method is sensitive,accurate,reliable and reproduc-ible,and can be used for the simultaneous determination of caffeic acid,isoschaftoside and schaftoside in Xiaoyan lidan tablet the quality control of Xiaoyan lidan tablet.

Study on Content Dynamic Changes of Three Water-soluble Flavonoids in Isodon Lophanthoides / 中国药师

Objective:To establish an HPLC method for the simultaneous determination of viceninⅡ, isoschaftoside and schafto-side in Isodon lophanthoides, and analyze their content dynamic changes. Methods:The HPLC system consisted of a Phenomenex luna C18(250 mm ×4.6 mm, 5 μm)column and a solution system of methanol and 0.5% formic acid with gradient elution, the flow rate was 0. 8 ml·min-1 and the detection wavelength was 334 nm at the column temperature of 25℃and the injection volume was 10 μl. Results:The linear range of viceninⅡ, isoschaftoside and schaftoside was 0.130-3.110 μg(r =0.999 8), 0.180-4.540 μg(r =0.999 9) and 0.080-1.970 μg(r =0. 9999), respectively. The average recovery was 96.8% (RSD =1.9%), 99.2% (RSD =1. 6%) and 97. 1% (RSD=1. 6%) (n=6), respectively. The above three kinds of water-soluble flavonoids from seedling stage to significant accumulation, isoschaftoside and schaftoside reached the highest value in March and August, the content of VitexinⅡ in January and April tends to be stable, in May began to increase gradually, reached the maximum value in August, then began to de-crease. Conclusion:With the contents of water-soluble flavonoids as the indicators, the best harvest time of Isodon lophanthoides is March and August, and the quality of the medicinal materials harvested in August is the best.

Simultaneous Determination of Schaftoside, Isoschaftoside, Deoxyelephantopin and 4, 5-Dicaffeoylquinic Acid in Shennongcha Granules by HPLC / 中国药师

Objective:To establish an HPLC method for determining four constituents ( schaftoside, isoschaftoside, deoxyelephan-topin and 4,5-dicaffeoylquinic acid) in Shennongcha granules. Methods:An HPLC method was performed on a Hypersil C18 column (200 mm × 4. 6 mm,5 μm) with the mobile phase of acetonitrile as the phase A and 0. 025 mol·L-1 phosphoric acid solution as the phase B with gradient elution. The flow rate was 1. 3 ml·min-1 . The detection wavelength was set at 270 nm for schaftoside and isos-chaftoside, 208 nm for deoxyelephantopin and 327 nm for 4, 5-dicaffeoylquinic acid. The column temperature was room temperature. Results:The calibration curve was linear over the concentration range of 5. 850-117. 000 μg·ml-1 for schaftoside, 4. 650-93. 000 μg ·ml-1 for isoschaftoside, 4. 160-83. 200 μg · ml-1 for deoxyelephantopin and 5. 470-109. 400 μg · ml-1 for 4, 5-dicaffeoylquinic acid. The correlation coefficient of all curves was more than 0.999. The average recoverywas 97.70% (RSD=1.40%), 96.87%(RSD=1.13%), 97.53%(RSD =1.69%) and 99.29%(RSD =1.01%) (n =6) , respectively. Conclusion: The developed HPLC method is simple,accurate,and can be used in the content determination of Shennongcha granules.

Study on the Content Dynamic Changes of Caffeic Acid,Vicenin-2 and Isoschaftoside in Rabdosia lophanthoi-des / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of caffeic acid,vicenin-2 and isoschaftoside in Rabdosia lophanthoides to assess its optimal harvest period. METHODS:HPLC was performed on the column of Diamonsil C18 with mobile phase of methanol-0.1% phosphoric acid solution (gradient elution) at a flow rate of 0.8 ml/min ,detection wave-length was 334 nm ,column temperature was 25 ℃ and volume injection was 15 μl. RESULTS:The linear range was 0.23-5.68 μg/ml for caffeic acid(r=0.999 9),0.31-7.76 μg/ml for vicenin-2(r=0.999 8)and 0.45-11.30 μg/ml for isoschaftoside(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%,average recoveries were 95.1%-98.9%(RSD=1.8%, n=6),97.4%-101.3%(RSD=1.7%,n=6) and 95.5%-98.8%(RSD=1.4%,n=6),respectively. The contents of above-men-tioned 3 ingredients of R. lophanthoides were the highest harvested in Apr. to May and Jul. to Aug. in a year. CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the simultaneous determination of caffeic acid,vicenin-2 and isoschaftoside in R. lophanthoides. The optimal harvest period is Apr. to May and Jul. to Aug. in a year.

Water-soluble constituents from Isodon lophanthoides var. gerardianus and their antitumor activities / 中草药

Objective: To investigate the water-soluble constituents of Isodon lophanthoides var. gerardianus. Methods: Compouds were isolated and purified by D-101, Sephadex LH-20, and ODS column chromatographies. The chemical structures were elucidated on the basis of spectral data and physicochemical properties. Results: Twelve compounds were isolated and identified as vitexin II (1), vicenin III (2), isoschaftoside (3), schaftoside (4), vitexin (5), 6, 8-di-C-α-L-arabinosylapigenin (6), apigenin-7-O-glucuronide (7), apigenin 6-C-β-L-arabin-osyl-8-C-α-L-arabinopyranoside (8), apigenin 6-C-β-D-xylopyranosyl-8-C-α-L-arabinopyranoside (9), caffeic acid (10), rosmarinic (11), and rutin (12). Conclusion: Compounds 1-9 are separated from I. lophanthoides var. gerardianus for the first time, and constituents of the water-soluble parts of I. lophanthoides var. gerardianus are studied systematically for the first time. Compounds 1, 6, and 11 exert antiproliferative effects on HepG2 cells.

Studies on reference substance of Guangjinqiancao Zonghuangtong Capsule / 中成药

AIM:To prepare a compound as the chemical reference substance of Guangjinqiancao Zonghuangtong Capsule.METHODS:To apply general column chromatography combined with preparative HPLC to isolate the target compound,to use analytic HPLC to determine the purity,stability and its content in the capsule,and to employ spectroscopic analysis (UV,IR,ESI-MS,1H-NMR,13C-NMR,DEPT,1H-13CCOSY,1H-1HCOSY,1DHOHAHA.1D.NOE,HMBC) to elucidate the structure of the isolated compound.RESULTS:The obtained compound was identified as isoschaftoside with the purity of over 99%, which was stable within 3 months at ambient temperature.As for isosehaftoside solution.it was stable within 8 h at ambient temperature.Its content in the capsule was above 3.0%.CONCLUSION:Isoschaftoside is a qualified reference substance for analytic assay ofGuangjinqiancao Zonghuangtong Capsule,and can be isolated from Desmodium styracifolium(Osb.)Merr.

Study on Identification of Rhizoma Arisaematis and Content Determination of Flavonoids / 中国药房

OBJECTIVE:To establish the identification method of Rhizoma Arisaematis and content determination of flavon-oids.METHODS:TLC was used to identify Rhizoma Arisaematis with schaftoside and ischaftoside as reference substances.The content of flavonoids was determined by HPLC.RESULTS:TLC of test sample and that of control substance had same color dots.The linear range of schaftoside and isoschaftoside were 7.925～126.8 ?g?mL-1(r=0.999 9) and 3.996～63.94 ?g?mL-1(r=0.999 7) respectively.Average recoveries were 99.4% for schaftoside (RSD=2.10%,n=9) and 99.52% for isoschaftoside(RSD=2.42%,n=9).CONCLUSION:The method is simple,accurate and reproducible for the quality evaluation of Rhizoma Arisaematis.