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This study aimed to characterize and identify the non-volatile components in Pogostemonis Herba by using ultra-perfor-mance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) combined with UNIFI and an in-house library. The chemical components in 50% methanol extract of Pogostemonis Herba were detected by UPLC-Q-TOF-MS in both positive and negative MS~E continuum modes. Then, the MS data were processed in UNIFI combined with an in-house library to automatically characterize the metabolites. Based on the multiple adduct ions, exact mass, diagnostic fragment ions, and peak intensity of compounds and the fragmentation pathways and retention behaviors of reference substances, the structures identified by UNIFI were further verified and those of the unidentified compounds were tentatively elucidated. A total of 120 compound structures were identified or tentatively identified, including flavonoids, phenylpropanoids, phenolic acids, terpenes, fatty acids, alkaloids, and phenylethanoid glycosides. Sixteen of them were accurately identified by comparison with reference substances, and 53 compounds were reported the first time for Pogostemonis Herba. This study systematically characterized and identified the non-volatile compounds in Pogostemonis Herba for the first time. The findings provide a scientific basis for revealing the pharmacodynamic material basis, establishing a quality control system, and developing products of Pogostemonis Herba.
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This study aims to identify the chemical constituents from Callicarpa kwangtungensis and determine their activities. MCI, ODS, and Sephadex LH-20 chromatography and semi-preparative HPLC were employed to separate the chemical constituents. A total of 15 compounds were separated, and their structures were identified on the basis of spectroscopic analysis and comparison with the data in relevant literature. Specifically, the 15 compounds were 3-O-α-L-rhamnopyranosyl-6-O-β-D-apiofuranosyl-4-O-E-caffeoyl-D-glucopyranoside(1), 3,6-O-α-L-dirhamnopyranosyl-4-O-E-caffeoyl-D-glucopyranoside(2), β-OH-forsythoside B(3), β-OH-poliumoside(4),(+)-lyoniresinol-3α-O-β-D-apiofuranosyl-(1→2)-β-D-glucopyranoside(5),(+)-lyoniresinol-3α-O-β-D-glucopyranoside(6),(-)-lyoniresinol-3α-O-β-D-glucopyranoside(7), kelampayoside A(8), descaffeoylpoliumoside(9), acteoside(10), alyssonoside(11), poliumoside(12), isacteoside(13), acetyl forsythoside B(14), and forsythoside B(15). Compounds 1 and 2 were novel, and the NMR data of compounds 3 and 4 were reported here for the first time. Furthermore, the hemostatic activities of the extract and abundant ingredients(compounds 12 and 15) of C. kwangtungensis were determined with Yunnan Baiyao as the positive control and normal saline as the negative control. The extract and compounds 12 and 15 significantly shortened the tail tip bleeding time in mice.
Subject(s)
Animals , Mice , Callicarpa , Hemostatics , China , Glycosides/chemistryABSTRACT
ObjectiveIn order to explore the changes of chemical constituents in Plantaginis Semen before and after stir-frying， ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MSE） was used to rapidly identify and semi-quantitatively analyze the differential components in Plantaginis Semen processed at different stir-frying time. MethodWaters ACQUITY UPLC BEH C18 column （2.1 mm×100 mm， 1.8 μm） was employed with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-1 min， 5%-10%B； 1-2 min， 10%-15%B； 2-10 min， 15%-20%B； 10-12 min， 20%-40%B； 12-13 min， 40%-100%B； 13-14 min， 100%-5%B； 14-15 min， 5%B）， the flow rate was 0.3 mL·min-1， the column temperature was 40 ℃， and the injection volume was 3 μL. Electrospray ionization （ESI） was applied for mass spectrometric analysis under positive and negative ion modes， and the scanning range was m/z 50-1 500. MarkerLynx 4.1 software was used to find the differential compounds， and the intensity of each ion peak in samples with different stir-frying time was compared to study the content variations of these compounds. ResultA total of 20 components with potential significant differences were found， among which 17 were identified and 3 were unknown， mainly including phenylethanoid glycosides， iridoid glycosides， alkaloids and others. After processing， the peak intensities of 7 compounds， such as sucrose， geniposidic acid， verbascoside and plantagoguanidinic acid A， in Plantaginis Semen decreased. The peak intensities of orobanchoside， dianthoside and plantain D increased first and then decreased during the stir-frying process. The peak intensities of 10 compounds （decaffeoylacteoside， calceolarioside A， isoacteoside， etc.） increased， and 9 of them were newly generated components. ConclusionThe content and composition of the chemical components in Plantaginis Semen changed significantly after stir-frying， which may be related to the reduction of laxative effect and the enhancement of antidiarrheal and diuretic activities of Plantaginis Semen after stir-frying.
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The purpose of the research is to study the bioactive constituents of Callicarpa nudiflora. From the 65% ethanol extract of C. nudiflora leaves, ten compounds were isolated by macroporous adsorption resin, Sephadex LH-20, ODS, silica gel, and preparative HPLC. These compounds were identified as callicapene M6(1), sterebin A(2), isomartynoside(3), crenatoside(4), luteolin-7-O-neohesperidoside(5), apigenin-7-O-β-D-neohesperidoside(6), isoacteoside(7), acteoside(8),(7R)-campneoside I(9), and(7S)-campneoside I(10) on the basis of NMR, HR-ESI-MS, and optical rotation data. Compound 1 was obtained as a new compound. Compounds 2 and 4 were isolated from the genus Callicarpa for the first time. Compounds 9 and 10 were isolated from C. nudiflora for the first time.
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Callicarpa , Chromatography, High Pressure Liquid , Diterpenes , Molecular Structure , Plant LeavesABSTRACT
Tanreqing Injection has the functions of clearing heat, resolving phlegm and detoxifying. It mainly contains amino acids, iridoids, flavonoids, phenylethanoid glycosides, steroids and other chemical components. Studies show that Tanreqing Injection has many pharmacological activities such as anti-inflammation, anti-bacteria, anti-virus, inhibition of liver injury and etc. Numerous domestic and foreign research efforts have been focused on the study of Tanreqing Injection. This paper comprehensively reviewed the recent progress in research on chemical constituents, pharmacological actions and clinical application of Tanreqing Injection, so as to provide some reference for its further study.
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Several Orobanche medicinal plants sometimes served as alternative sources of Cistanches Herba, attributing to the benefits such as tonifying kidney, strengthening tendons and bones. Among them, O. coerulescens, O. cernua and O. pycnostachya have been widely utilized in northern China for treatments of pains in the loins and knees, impotence, and spermatorrhea. However, their chemical profiles haven't been elucidated. In the present study, UHPLC-IT-TOF-MS was implemented to conduct in-depth chemome profiling of O. coerulescens, O. cernua and O. pycnostachya, aiming to achieve a comprehensive chemical characterization and to provide pronounced information for the quality control and clinical applications. An ACE Ultra-Core 2.5 Super C_(18)(3.0 mm×150 mm, 2.5 μm) column was deployed for chromatographic separations, and high-resolution MS~n spectra were recorded by IT-TOF-MS. Forty-eight components, in total, were observed, and thirty-eight ones were structurally annotated according to proposing mass fragmentation patterns, matching with relevant databases. Particularly, nine ones were confirmed by reference compounds. Overall, the chemical compositions of O. coerulescens and O. cernua are quite similar, and differences occur between O. pycnostachya and the prior two ones; primary chemical family is phenylethanoid glycosides, and several lignan glycosides as well as iridoid glycosides are also observed; the primary components include acteoside, isoacteoside, crenatoside and 2'-acetylacteoside, etc.
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Male , China , Cistanche , Glycosides , Orobanche , Plants, MedicinalABSTRACT
Objective::To study the chemical constituents in Baihe Dihuangtang by ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). Method::The separation was performed on an ACQUITY UPLC BEH C18 column (2.1 mm×50 mm, 1.7 μm) by a gradient elution of acetonitrile-0.1%formic acid solution. The flow rate was 0.4 mL·min-1 and the column temperature was 30 ℃, the injection volume was 5 μL. Electrospray ionization was applied and the data were collected via positive and negative ion modes. By using SCIEX OS 1.4 software, the chemical constituents were analyzed based on the retention time, excimer ion peak and fragment ion peak of the compounds, as well as comparison with reference substances and literature data. Result::A total of 49 chemical constituents in Baihe Dihuangtang were identified, including 5 phenolic glycerides, 15 phenylethanoid glycosides, 11 iridoids, 8 lonones, 3 phenylpropanoids, 2 nucleosides, 1 organic acid, and 4 other compounds. Among them, phenolic glycerides belonged to Lilii Bulbus, and other components mainly belonged to Rehmanniae Radix. Four chemical constituents (acteoside, isoacteoside, ferulic acid and caffeic acid) were identified by comparison of reference substances. Conclusion::The established detection method can quickly and accurately analyze the chemical constituents of Baihe Dihuangtang. The information of chemical constituents in Baihe Dihuangtang is comprehensively expounded. The study establishes a foundation for the research of quality control, material foundation of efficacy and the development of compound preparations of Baihe Dihuangtang.
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In order to improve the quality control level of Ligustri Lucidi Fructus(LLF) and to explore the changes of chemical components after processing,the HPLC method for fingerprint and simultaneous determination of the major polar components in LLF were established. The octadecylsilane bonded silica gel was used as the stationary phase,with acetonitrile as the mobile phase A and0. 2% formic acid as the mobile phase B in a gradient elution procedure at a flow rate of 1. 0 m L·min-1. The detection wavelength was set at 280 nm and the column temperature was 25 ℃. There were 22 common peaks,20 of which were selected from the fingerprint of LLF and its wine-steamed product,respectively,and 14 chromatographic peaks were identified with reference substances. With the same chromatographic conditions,seven components were quantitatively analyzed and the results of system adaptability and methodology investigation all met the requirements of content determination. Compared with the crude LLF,the content of 5-hydroxymethyl furfural and salidroside significantly increased in wine-steamed LLF,while the contents of iridoid glycosides generally decreased. The method provided a basis for quality control of LLF and its processed products as well as the related preparations.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Fruit , Chemistry , Furaldehyde , Glucosides , Iridoid Glycosides , Ligustrum , Chemistry , Phenols , PhytochemicalsABSTRACT
Objective: To establish the plant tissue culture system of Cistanche tubulosa, and determine the effect of drought stress on accumulation of two respective phenylethanoid glycosides in it. Methods The major chemical constituents of C. tubulosa by tissue culture were analyzed by HPLC-UV and HR-MS. The cell growth curves were also determined. In addition, the effects of drought stress on the phenylethanoid glycosides (echinacoside and acteoside) content in the tissue culture system of C. tubulosa were also studied by using NaCl, mannitol and PEG6000 as osmotic regulators, respectively. Results:Chemical constituents analyses revealed that callus and suspension cultures of C. tubulosa could produce the respective phenylethanoid glycosides of echinacoside and acteoside as in wild plant; Cell growth curves indicated that 30 d were the optimum culture period of callus culture; The cell growth rate and the accumulation of echinacoside and acteoside were mostly inhibited when the callus cells were under drought stress induced by NaCl or mannitol. Meanwhile, the accumulation of echinacoside and acteoside in cell suspension culture of C. tubulosa could be effectively enhanced by treatment with PEG6000. The maximum biomass of echinacoside and acteoside could reach to (1.07 ± 0.10) g/L and (0.12 ± 0.01) g/L 15 d after induction, respectively. And their contents were 20.94% and 2.27% separately based on the cell dry weight (DW) after 15 d of treatment with 6% PEG6000, which were 1.29 and 1.19 fold higher than the control group. Conclusion:Drought stress induced by PEG6000 could effectively enhance the accumulation of echinacoside and acteoside in cell suspension culture of C. tubulosa.
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Phenylethanoid glycosides (PeGs) are natural active ingredients of many plants at home and abroad. They possess a spectrum of beneficial activities, such as anti-oxidant, hepatoprotective, whitening, and neuroprotective. However, the content of PeGs in food or medicinal materials is low and unstable. During the past decade, studies on biosynthesis and metabolic regulation of PeGs have been extensively carried out. Here, the recent achievements in biosynthesis and metabolic regulation of PeGs in plants and microorganisms are reviewed, as well as in total synthesis and semi-synthesis of PeGs. Hopefully, this work done so far will provide reference for elucidating the biosynthesis mechanism of PeGs, stabilizing and improving the content of PeGs in herbal materials, improving the quality, and synthesizing PeGs by microbial metabolic engineering or chemical synthesis.
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Objective: To investigate the adsorption kinetics and thermodynamic characteristics of phenylethanoid glycosides (PGs) in the leaves of Callicarpa nudiflora on macroporous resins and provide a reference for the separation and purification of these compounds. Methods: With adsorption and desorption ratio as indexes, optimum types of macroporous resin for purification of PGs were selected from 8 kinds of macroporous resins by static adsorption and desorption tests, and then adsorption kinetics model and adsorption isotherm model of PGs were established to investigate their adsorption processes. Results: SP-825 and SP-207 resin were selected and they have similar adsorption process for PGs. Both of them showed a fast adsorption in 0-60 min, a slow adsorption in 60-360 min, and an equilibrium adsorption stage after 360 min. Adsorption dynamic behavior was well described by quasi-second-order equation of both SP-825 and SP-207 macroporous resins, and adsorption rate was mainly controlled by liquid film diffusion and intraparticle diffusion. Equilibrium adsorption data fitted Langmuir and Freundlich isotherm equations well. Both of the two kinds of resins showed good adsorption properties for PGs, and the adsorption process belongs to favorable adsorption. Conclusions: Both of the kinetic model and thermodynamic model can well describe the adsorption process of SP-825 and SP-207 macroporous resins and the two resins were regarded as excellent adsorption resins for the purification of PGs from the leaves of Callicarpa nudiflora.
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OBJECTIVE： To establish a method for rapid determination of total phenylethanoid glycosides and total iridoid glycosides in the root of Rehmannia glutinosa. METHODS： The contents of total phenylethanoid glycosides and total iridoid glycosides in medicinal material samples were determined by UV spectrophotometry. Quantitative model of total phenylethanoid glycosides and total iridoid glycosides in medicinal samples was established by NIRS-PLS method. The optimal pretreatment spectra were multivariate scattering correction combined with first derivative method， standard normalization combined with first derivative method. The optimum spectral ranged from 6 703.35-11 065.54 cm－1 and 3 999.63-9 102.36 cm－1. The optimum principal factor number were 10 and 7. RESULTS： The content determination of total phenylethanoid glycosides and total iridoid glycosides in medicinal material samples was proved to meet the requirements by methodological experience. The internal cross validation determination coefficients of total phenylethanoid glycosides and total iridoid glycosides were 0.998 2 and 0.980 9. The correction of root mean square error was 0.032 7 and 0.186 0. The root mean square error of prediction were 0.035 5 and 0.035 1. The root mean square error of cross validation were 0.256 9 and 0.574 3. The predicted values of total phenylethanol glycosides and total iridoid glycosides were 0.268％-1.636％ and 3.424％-6.978％， respectively； the determination value of them were 0.299％-1.629％ and 3.431％-6.952％， respectively； the absolute deviations were －0.042％-0.067％ and －0.111％-0.088％， respectively；the relative deviations were －0.819％-0.076％、－2.257％-1.672％， respectively；There was no statistical significance between predicted values and measured values （P＞0.05）. CONCLUSIONS： The method is accurate and simple. The method can be used for the rapid determination of total phenylethanoid glycosides and total iridoid glycosides in different germplasms of R. glutinosa.
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The genus Orobanche, Cistanche and Boschniakia are taxonomically classified as members of Orobanchaceae. The medicinal plants of these three genera are closely related, and their traditional curative effects are similar. As representative compounds, phenethyl glycosides are predominantly dominant type both in amount and in chemical structural varieties, which are considered to be the important bioactive material basis of these genera. In this paper, phenethyl glycosides and their pharmacological activities are described in a single list. In addition, the other compounds were also reviewed in order to better compare the difference of the bioactive substances. These findings have important reference value for effective development and rational utilization of resources of medicinal plants in the family Orobanchaceae.
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Cistanche , Glycosides , Orobanchaceae , Orobanche , Plants, MedicinalABSTRACT
Echinacoside (ECH) is one of the active ingredients in Cistanche Herba and the principal effective component of Memoregain© as well. Moreover, a new agent namely Naoqing Zhiming tablet, derived from ECH has been licensed for clinical trials. However, the knowledge regarding the stability of is limited, till now, initiating a significant barrier for its further development along with the clinical trials. Herein, we aim to in depth characterize the transformation pattern of ECH in methanol. When ECH was stored in methanol, two primary products (P1 and P2) could be observed in HPLC chromatogram. A home-made automated fraction collector was configured via employing two 2-phase/6-port electronic valves to prepare P1 and P2. Following ¹H-NMR and LC-MS/MS assays, P1 and P2 were unambiguously identified as acteoside and cistanoside A, respectively. Moreover, the existences of cis-ECH, cis-acteoside, and cis-cistanoside A were claimed after careful analysis of the ¹H-NMR spectra of ECH, P1 and P2. Above all, the primary transformation pathways of ECH in methanol included methylation as well as hydrolysis, and mild transformation could also be initiated by cis/trans- configuration transferring for the caffeoyl group. The findings obtained in current study are envisioned to provide useful insight for the further development of ECH and the impurity detection of Naoqing Zhiming tablet. Moreover, the automated fraction collector configured in current study is able to serve as a versatile tool for the collection of signals-of-interest within phytochemical evaluations and impurity isolation.
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As a holoparasitic plant, Cistanche deserticola is one of the two original sources of Cistanches Herba that is one of the most famous tonic medicines, in Chinese Pharmacopoeia. The succulent stems are used for medicinal usage, whereas those lignified stems as well as flowers of less pharmacological importance are usually deserted, suggesting extensive resource waste. Herein, chemical characterization of the flowers along with lignified stems was conducted using HPLC-IT-TOF-MS aiming to explore the medicinal valu of those non-medicinal parts. Following ultrasonication-assisted extraction with 50% aqueous methanol, either flower or lignified stem extract was subjected onto LC-IT-TOF-MS equipped with a Capcell core ADME column to acquire both MS¹ and MSº spectra, and gradient elution was carried out with combinatory 0.1% aqueous formic acid and acetonitrile. Both positive and negative ionization polarities were deployed, resulting in the observation of 62 components, in total. Thirty-nine signals were structurally annotated, including phenylethanoid glycosides, iridoids, lignans and saccharides according to matching with authentic components and literature information, as well as applying the proposed mass fragmentation rules. A total of 62 ones were putatively identified. Above all, lignified stems and flowers should not the qualified substitutes for the succulent stems attributing to the significant differences between the medicinal portion and those parts with less medicinal values.
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The chemical constituents from the ethanol extract of Clerodendrum japonicum were isolated by a combination of various chromatographic techniques including column chromatography over silica gel, sephadex LH-20, ODS and reversed phase HPLC. Sixteen compounds with a pair of epimers were elucidated through the application of physicochemical properties with modern spectral analysis technology as 7α-hydroxy syringaresinol (1), (-)-syringaresinol (2), (-)-medioresinol (3), 2″,3″--acetylmartyonside (4), 2″--acetyl-martyonside (5), martinoside (6), monoacetyl martinoside (7)，cytochalasin O (8), 9-epi-blumenol B (9), (6R, 9S) and (6R,9)-9-hydroxy-4-megastigmen-3-one (10a，10b), (6R,9S)-3-oxo-α-ionol (11), (-)-dehydrovomifoliol (12)，megastigm-5-en-3,9-diol (13), (3R,6E,10S)-2,6,10-trimethyl-3-hydroxydodeca-6,11-diene-2,10-diol (14), (2)-butylitaconic acid (15), 3-(3&-hydroxybutyl)-2,4,4-trimethylcyclohexa-2,5-dienone (16), (-)-loliolide (17), of which compound 1 and 15 are new natural product, the other compounds were isolated for the first time from Clerodendrum japonicum except for compounds 4, 6 and 7.
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Objective Cistanches Herba is a kind of tonic traditional Chinese medicine with several therapy effects including tonifying kidney-yin, anti-dementia, anti-aging and relaxing bowel. Phenylethanoid glycosides (PhGs) are the major effective components in C. tubulosa. However, there were no further studies on molecular pharmacologic mechanisms due to its complex components and mechanism diversity of action in PhGs till to now. The aim of this study was to investigate the target protein groups and related mechanisms associated with PhGs in anticerebral ischemia-reperfusion injury. Methods The middle cerebral artery occlusion (MCAO) model was established in rats, and the protective effects of PhGs on cerebral ischemia-induced injuries were determined. A kind of solid bead whose surface was cross-linked with PhGs was prepared to capture the target proteins from brain tissue lysates. The target proteins were further identified with LC-MS/MS. Results PhGs significantly inhibited cerebral ischemia-induced injuries by reducing ischemia size and rat behavioral scores and elevated the SOD levels in rat brain tissues. Eighteen target proteins were identified based on “target fishing” strategy and divided into 9 kinds according to their biological functions, including anti-oxidation, ion channel, immunoregulation, cell survival and cytoskeleton, etc. Conclusion These findings reveal the potential pharmacological mechanisms of PhGs in anti-dementia, fatigability alleviating, anti-tumor, immunoloregulation, etc, and also present a promising technology for investigating the complicated pharmacological mechanisms of traditional Chinese medicine.
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The constituents of the whole plant of Orobanche coerulescens were isolated and purified by using various column chromatographic techniques including D101, silica gel and ODS. The structures were identified by spectroscopic analyses including NMR and MS. A new phenylethanol glycoside was isolated from the whole plant of O. coerulescens, and was identified as 2-(3-methoxy-4-hydroxyphenyl)-ethanol-1-O- [(1→3)-O-α-L-rhamnopyranosyl-4, 6-O-di-feruloyl]-β-D-glucopyranoside, named as orobancheoside B. Through the antibacterial activity test, orobancheoside B was proved to have certain antibacterial activity, and be one of the main active components of O. coerulescens. The research result will laid a foundation for the medicinal materials and quality control research. Activity screening, broomrape orobancheoside B has certain antibacterial activity, as one of the main active components of O. coerulescens, and to constantly improve the quality of the medicinal materials laid a foundation.
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Objective To establish a method for determination of phenylethanoid glycoside and acteoside in Plantago Herba. Methods UV-visible spectrophotometric method was used for the determination of the content of phenylethanoid glycosides compounds in Plantago Herba. HPLC method was used for the determination of acteoside in Plantago Herba. Chromatographic column with C18 ODS2 (4.6 mm × 150 mm, 5 μm) was used. Acetonitrile-0.1%formic acid (13:87) was as mobile phase; the flow rate was 1 mL/min; the detection wavelength was 332 nm; the column temperature was 30 ℃; the sample volume was 10 μL. Results The contents of phenylethanoid glycoside in Plantago Herba from different producing areas were among 1.03%–3.47%. Acteoside with peak area over the 0.0062–1.55 mg range showed a good linear relationship; the sample recovery rate was 98.9%, and the RSD was 1.6%. The contents of acteoside in Plantago Herba from different producing areas was among 0.18%–0.56%. Conclusion The method is simple, stable and reproducible, which can be used for the determination of phenylethanoid glycoside and acteoside in Plantago Herba from different producing areas and provide experimental basis for quality control of Plantago Herba.
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The aim of this study was to evaluate the seed quality of C.deserticola and establish quality grading rules of seeds by detecting the impacts of different processing methods on the contents of the effective components of C.deserticola for optimizing the suitable processing method.The seed quality was judged by thousand-kernel weight,empty embryo rate and water content.The samples were preliminary processed by freeze-drying,natural drying and hot air circulation drying respectively,and the content of phenylethanoid glycosides was determined by HPLC-UV.The seed quality classification standard of C.deserticola was established,and the seeds were divided into three grades based on the standard.It was found that freeze-drying method was optimum,featuring less effective component loss,beautiful appearance of herbal piece,crisp texture and fast drying.In conclusion,this study laid a foundation for the quality control of the seeds of C.deserticola with the provision of scientific evidence for the initial processing of the fresh product.