Tubuloside B

Integrated Strategy Drives Direct Infusion-Tandem Mass Spectrometry as an Eligible Tool for Shotgun Pseudo-Targeted Metabolomics of Medicinal Plants.[Pubmed:33439008]

Anal Chem. 2021 Feb 2;93(4):2541-2550.

Direct infusion (DI) has an extraordinary high-throughput advantage. Pseudo-targeted metabolomics (PTM) has been demonstrated integrating the merits of both nontargeted and targeted metabolomics. Herein, we attempted to implant DI into the PTM concept to configure a new strategy allowing shotgun PTM. First, a versatile MS/MSALL program was applied to acquire MS(1) and MS(2) spectra. Second, online energy-resolved MS (online ER-MS) was conducted to obtain breakdown graph as well as optimal collision energy (OCE) for each ion transition paired by precursor ion and the dominant product ion. Third, selected reaction monitoring (SRM) was responsible to output a quantitative dataset with a constant length. Moreover, breakdown graph also served as orthogonal structural evidence when matching MS(2) spectra between DI-MS/MS and an in-house library to strengthen structural annotation confidence. To evaluate and illustrate the utility of the new strategy toward shotgun PTM of medicinal plants, in-depth chemome comparison was conducted within three Cistanche species, all of which are edible medicinal plants and playing essential roles for turning the deserts into the oases. A total of 185 variables participated in the quantitative measurement program. Each diagnostic ion pair was featured with an OCE. Significant species differences occurred, and echinacoside, acteoside, isoacteoside, 2'-acetyl-acteoside, Tubuloside B, mannitol, sucrose, betaine, malate, as well as choline were found to be confirmative chemical markers offering primary contributions toward the species discrimination. After cross-validation with LC-MS/MS, DI-MS/MS fortified with the new strategy is an eligible tool for shotgun PTM, beyond Cistanche plants.

[Comparative studies of three Cistanche speices based on UPLC specific chromatogram and determination of main components].[Pubmed:31602949]

Zhongguo Zhong Yao Za Zhi. 2019 Sep;44(17):3749-3757.

Based on UPLC specific chromatogram and determination of seven main components,this study aimed at evaluating the quality of Cistanche deserticola,C. tubulosa and C. sinensis. Echinacoside,cistanoside A,verbascoside,tubuloside A,isoacteoside,2'-acetylacteoside,Tubuloside B were used as reference substances. UPLC analysis was performed on a Waters ACQUITY UPLC HSS T3 column( 2. 1 mmx100 mm,1. 8 mum). The mobile phase was acetonitrile-0. 08% trifluoroacetic acid solution. The flow rate was0. 3 mL.min-1,and the injection amount was 10 muL. The column temperature was 40 ,and the detection wavelength was 330 nm.The UPLC specific chromatograms were processed with ChemPattern software. UPLC specific chromatograms of C. deserticola and C.tubulosa from different samples were of high similarity,but the similarities of their counterfeit C. sinensis were less than 0. 06. Both of cluster and principal component analysis can distinguish certified products and counterfeits. The content ratios of echinacoside/verbascoside and verbascoside/isoacteoside were quite different between C. deserticola and C. tubulosa,which had distinct significance.The UPLC specific chromatogram and contents of seven main components can provide a basis for quality evaluation of Cistanches Herba.

Determination of tubuloside B by LC-MS/MS and its application to a pharmacokinetic study in rats.[Pubmed:29143972]

Biomed Chromatogr. 2018 Apr;32(4).

Tubuloside B, a novel neuroprotective phenylethanoid, is a major active constituent of Cistanche tubulosa and Cistanche deserticola. A specific and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantification of Tubuloside B in rat plasma. Sample preparation was conducted through a protein-precipitation extraction with methanol using tubuloside A as internal standard (IS). Chromatographic separation was achieved using a Capcell Pak C18 column (2.0 x 50 mm, 5 mum) with a mobile phase of methanol-10 mm ammonium acetate buffer (70:30, v/v) in an isocratic elution. Mass spectrometry analysis was performed in negative ionization mode with selected reaction monitoring transitions at m/z 665.1 --> 160.9 for Tubuloside B, and m/z 827.1 --> 160.9 for IS. Calibration curves were linear over the range of 1.64-1640 ng/mL for plasma samples samples (R(2) > 0.990). The lower limit of quantification (LLOQ) was 1.64 ng/mL. The intra- and inter-day accuracy was between 92.3 and 113.0% with the RSD <9.23% at all LLOQ and quality control levels. Finally, this method was successfully applied in the pharmacokinetics study of Tubuloside B after intravenous administration.

[Caffeoyl phenylethanoid glycosides from Callicarpa kwangtungensis].[Pubmed:25095374]

Zhongguo Zhong Yao Za Zhi. 2014 May;39(9):1630-4.

Phytochemical investigation on the EtOH extract from the aerial part of Callicarpa kwangtungensis led to the isolation and characterization of 10 caffeoyl phenylethanoid glycosides, 2'-acetylacteoside (1), tubuloside E (2), acteoside (3), Tubuloside B (4), isoacteoside (5), alyssonoside (6), 2'-acetylforsythoside B (7), brandioside (8), forsythoside B (9), and poliumoside (10). Compound 4 was isolated from the plants of Verbenaceae,and 6 was obtained from the Callicarpa genus, for the first time, while compounds 1, 2, 5 and 7 were firstly reported from the plant.

Two-phase hollow fiber liquid phase microextraction based on magnetofluid for simultaneous determination of Echinacoside, Tubuloside B, Acteoside and Isoacteoside in rat plasma after oral administration of Cistanche salsa extract by high performance liquid chromatography.[Pubmed:24531006]

J Pharm Biomed Anal. 2014 Jun;94:30-5.

A new and fast sample preparation technique based on two-phase hollow fiber liquid phase microextraction (HF-LPME) with magnetofluid was developed to quantitate and determine the four phenylethanoid glycosides (PhGs) (Echinacoside, Tubuloside B, Acteoside and Isoacteoside) in plasma after oral administration of Cistanche salsa extract. Analysis was accomplished by reversed-phase high performance liquid chromatography (RP-HPLC) with ultraviolet detection. Parameters that affect the HF-LPME processes, such as the content of magnetic powder, the solvent type, salt content, stirring speed, extraction time and hollow fiber length, were investigated and optimized. Under the optimized conditions, the preconcentration factors for PhGs were higher than 625. The calibration curve for PhGs was linear in the range of 0.1-100ngmL(-1) with correlation coefficients greater than 0.9996. The intra-day and inter-day precision (RSD) were below 8.74% and the limits of detection (LOD) for the four PhGs were 8-15pgmL(-1) (S/N=3). The validated method was successfully applied to separate and determine the four PhGs in rat plasma after oral administration of C. salsa extract.

Simultaneous separation and determination of four phenylethanoid glycosides in rat plasma sample after oral administration of Cistanche salsa extract by microemulsion liquid chromatography.[Pubmed:24508672]

J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Mar 1;951-952:24-31.

A simple, rapid and specific method was developed to separate as well as to determine the four phenylethanoid glycosides (PhGs) (echinacoside, Tubuloside B, acteoside and isoacteoside) in rat plasma after oral administration of Cistanche salsa extract by reversed phase high performance liquid chromatography using a microemulsion as the mobile phase. The separations were performed on a Zorbax Extend-C18 column at 25 degrees C. Photodiode-array detector was conducted at 322nm and with a flow rate of 0.8mLmin(-1). The optimized microemulsion mobile phase consisted of 0.3% triethylamine in 20mM phosphoric acid at pH 6.0, 0.8% (v/v) ethyl acetate as oil phase, 1.5% (v/v) Genapol X-080 as surfactant, 2.5% (v/v) n-propanol as co-surfactant. Under the optimal conditions, the calibration curve for four PhGs was linear in the range of 10-1000ngmL(-1) with the correlation coefficients greater than 0.9994. The intra-day and inter-day precision (RSD) were below 8.64% and the limits of detection (LOD) for the four PhGs were 0.4-1.3ngmL(-1) (S/N=3). The microemulsion liquid chromatography (MELC) method was successfully applied to separate and determine the four PhGs in rat plasma after oral administration of C. salsa extract.

[Structure-activity relationships of phenylethanoid glycosides in plants of Cistanche salsa on antioxidative activity].[Pubmed:19873735]

Zhong Yao Cai. 2009 Jul;32(7):1067-9.

OBJECTIVE: To study the structure-activity relationships of phenylethanoid glycosides in plants of Cistanche salsa on antioxidative activity. METHODS: By the assay systems of DPPH*, the antioxidant activity of six phenylethanoid glycosides from plants of Cistanche salsa was determined to investigate the relationship between the antioxidant activities and phenylethanoid glycosides's structural characteristics. RESULTS: The antioxidative activity of phenylethanoid glycosides was variant with dose-dependent effect. The sequence of the strength of the antioxidative activity of the six components was shown to be 2'-Acetylacteoside > Acteoside > or = Tubuloside B > or = Isoacteoside > Echinacoside > Cistanoside A. CONCLUSION: The antioxidative activity of phenylethanoid glycosides is related to the number of phenolic hydroxyl, steric hindrance, 2-acetyl on the middle glucopyranose, and the location of phenolic hydroxyl. Additionally, it may be related to the alpha, beta-unsaturated ketone of phenl-2-propenoyl.

Protective effect of tubuloside B on TNFalpha-induced apoptosis in neuronal cells.[Pubmed:15456528]

Acta Pharmacol Sin. 2004 Oct;25(10):1276-84.

AIM: To investigate the neuroprotective effect of Tubuloside B, one of the phenylethanoids isolated from the stems of Cistanche salsa, on tumor necrosis factor-alpha (TNFalpha)-induced apoptosis in SH-SY5Y neuronal cells. METHODS: Cell viability was analyzed using MTT assay. Apoptotic cells were detected using Hoechst33342 staining, and confirmed by DNA fragmentation and flow cytometric analysis. The activity of caspase-3 was measured with special assay kit. The concentration of free intracellular calcium was determined with the probe Indo-1 by spectrometer. The level of intracellular reactive oxygen species and the potential of mitochondrial membrane were determined by laser scanning confocal microscopy (LSCM) combined with fluorescence probe H2DCFDA or JC-1 respectively. RESULTS: SH-SY5Y cells treated with TNFalpha 100 microg/L for 36 h showed typical morphological changes of apoptosis. DNA ladder could be observed by agarose gel electrophoresis. The highest percentage of apoptotic cells accumulated to 37.5 %. Following 36 h treatment with TNFalpha, accumulation of intracellular ROS and [Ca2+]i and decrease in mitochondrial membrane potential were observed, and caspase-3 activity increased by about five-fold compared with controls. However, pretreatment with Tubuloside B (1, 10, or 100 mg/L) for 2 h attenuated the TNFalpha-mediated apoptosis. The antiapoptotic action of Tubuloside B was partially dependent on an anti-oxidative stress effects, maintain of mitochondria function, decrease of concentration of free intracellular calcium and inhibition of caspase-3 activity. CONCLUSION: Tubuloside B has the neuroprotective capacity to antagonize TNFalpha-induced apoptosis in SH-SY5Y cells and may be useful in treating some neurodegenerative diseases.

Tubuloside B from Cistanche salsa rescues the PC12 neuronal cells from 1-methyl-4-phenylpyridinium ion-induced apoptosis and oxidative stress.[Pubmed:12451484]

Planta Med. 2002 Nov;68(11):966-70.

The neuroprotective effects of Tubuloside B, one of the phenylethanoids isolated from the Chinese herbal medicine Cistanche salsa, on 1-methyl-4-phenylpyridinium ion (MPP +)-induced apoptosis and oxidative stress in PC12 neuronal cells were investigated. PC12 cells treated with MPP + underwent apoptotic death as determined by MTT assay, flow cytometry and DNA agarose gel electrophoresis; intracellular accumulation of reactive oxygen species (ROS) was measured by DCFH-DA staining with laser scanning confocal microscopy (LSCM). Simultaneous treatment with Tubuloside B markedly attenuated MPP +-induced cytotoxicity, DNA fragmentation, and intracellular accumulation of ROS. These results strongly indicate that Tubuloside B prevents MPP +-induced apoptosis and oxidative stress. Tubuloside B may be applied as an antiparkinsonian agent.

Inhibition of nitric oxide by phenylethanoids in activated macrophages.[Pubmed:10913595]

Eur J Pharmacol. 2000 Jul 14;400(1):137-44.

Nitric oxide (NO) is one of the pro-inflammatory molecules. Some phenylethanoids have been previously shown to possess anti-inflammatory effects. Seven phenylethanoids from the stems of Cistanche deserticola, viz. isoacteoside, Tubuloside B, acteoside, 2'-O-acetylacteoside, echinacoside, cistanoside A and tubuloside A, were tested for their effect on NO radical generation by activated murine macrophages. At the concentration of 100-200 microM, all the phenylethanoids reduced (6.3-62.3%) nitrite accumulation in lipopolysaccharide (0.1 microgram/ml)-stimulated J774.1 cells. At 200 microM, they inhibited by 32.2-72.4% nitrite accumulation induced by lipopolysaccharide (0.1 microgram/ml)/interferon-gamma (100 U/ml) in mouse peritoneal exudate macrophages. However, these compounds did not affect the expression of inducible nitric oxide (iNOS) mRNA, the iNOS protein level, or the iNOS activity in lipopolysaccharide-stimulated J774.1 cells. Instead, they showed a clear scavenging effect (6.9-43.9%) at the low concentrations of 2-10 microM of about 12 microM nitrite generated from an NO donor, 1-propanamine-3-hydroxy-2-nitroso-1-propylhydrazino (PAPA NONOate). These results indicate that the phenylethanoids have NO radical-scavenging activity, which possibly contributes to their anti-inflammatory effects.

Hepatoprotective activity of phenylethanoids from Cistanche deserticola.[Pubmed:9525102]

Planta Med. 1998 Mar;64(2):120-5.

Four phenylethanoids isolated from the stems of Cistanche deserticola, acteoside (1), 2'-acetylacteoside (2), isoacteoside (3) and Tubuloside B (4), significantly suppressed NADPH/CCl4-induced lipid peroxidation in rat liver microsomes. Addition of them to primary cultured rat hepatocytes efficiently prevented cell damage induced by exposure to CCl4 or D-galactosamine (D-GalN). Acteoside (1) further showed pronounced anti-hepatotoxic activity against CCl4 in vivo.

Antioxidative effects of phenylethanoids from Cistanche deserticola.[Pubmed:8996643]

Biol Pharm Bull. 1996 Dec;19(12):1580-5.

The acetone-H2O (9:1) extract from the stem of Cistanche deserticola showed a strong free radical scavenging activity. Nine major phenylethanoid compounds were isolated from this extract. They were identified by NMR as acteoside, isoacteoside, 2'-acetylacteoside, Tubuloside B, echinacoside, tubuloside A, syringalide A 3'-alpha-rhamnopyranoside, cistanoside A and cistanoside F. All of these compounds showed stronger free radical scavenging activities than alpha-tocopherol on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and xanthine/xanthine oxidase (XOD) generated superoxide anion radical (O2-.). Among the nine compounds, isoacteoside and Tubuloside B, whose caffeoyl moiety is at 6'-position of the glucose, showed an inhibitory effect on XOD. We further studied the effects of these phenylethanoids on the lipid peroxidation in rat liver microsomes induced by enzymatic and non-enzymatic methods. As expected, each of them exhibited significant inhibition on both ascorbic acid/Fe2+ and ADP/NADPH/Fe3+ induced lipid peroxidation in rat liver microsomes, which were more potent than alpha-tocopherol of caffeic acid. The antioxidative effect was found to be potentiated by an increase in the number of phenolic hydroxyl groups in the molecule.