Spinacetin

UHPLC-HRMS based flavonoid profiling of the aerial parts of Chenopodium foliosum Asch. (Amaranthaceae).[Pubmed:31711317]

Nat Prod Res. 2019 Nov 12:1-5.

Chenopodium foliosum Asch. has been recognised by Bulgarian legislation as a medicinal plant. The decoction of its aerial parts has been used for treatment of cancer, as an immunostimulant and antioxidant drug. An UHPLC-HRMS profiling method was used for a comprehensive study of flavonoid composition of C. foliosum. Fourty flavonoid glycosides with nine aglycones (patuletin, gomphrenol, Spinacetin, 6-methoxykaempferol, kaempferol, quercetin, isorhamnetin, 3,5,3',4'-tetrahydroxy-6,7-methylenedioxyflavone and 3,5,4'-trihydroxy-3'-methoxy-6,7-methylenedioxyflavone) were detected. Kaempferol, quercetin and isorhamnetin glycosides were identified as minor components. A pseudo MS(3) experiment aided at discriminating 6-methoxykaempferol and isorhamnetin glycosides. Flavonoid composition dominated by di-, triglycosides and acylated flavonoids. Acid hydrolysis and GS-MS analysis confirmed the presence of D-glucose, D-apiose and L-rhamnose. Ten flavonoids were reported here for the first time.

Comparison of flavonoid and policosanol profiles in Korean winter-spinach (Spinacia oleracea L.) cultivated in different regions.[Pubmed:30611481]

Food Chem. 2019 May 1;279:202-208.

Spinach intake has long been highlighted globally because of its outstanding nutritional aspects. In this study, changes in flavonoids, a representative functional phytochemical group, were investigated by UPLC-QTof MS with multivariate analysis of winter-spinach samples from three different cultivation regions in Korea. From the partial least squares discriminant analysis (PLS-DA), the differences of flavonoids among the geographic locations were clearly distinguished. Seven spinach flavonoids (2, patuletin-3-O-glucosyl-(1-->6)-glucoside; 4, Spinacetin-3-O-glucosyl-(1-->6)-[apiosyl-(1-->2)]-glucoside; 8, patuletin 3-O-(2''-feruloylglucosyl)-(1-->6)-[apiosyl-(1-->2)]-glucoside; 11, Spinacetin 3-O-(2''-feruloylglucosyl)-(1-->6)-[apiosyl-(1-->2)]-glucoside; 12, patuletin 3-O-(2''-feruloylglucosyl)-(1-->6)-glucoside; 18, 5,3',4'-trihydroxy-3-methoxy-6:7-methylendioxyflavone-4'-glucuronide; 20, 5,4'-dihydroxy-3,3'-dimethoxy-6:7-methylendioxyflavone-4'-glucuronide) were evaluated as key markers among 20 isolated metabolites. Interestingly, the contents of individual marker were significantly different among the groups, though total amount of flavonoids were almost same. Additionally, policosanols (PCs) in the winter-spinach was examined quantitatively using GC-MS for the first time. The PCs were analyzed as the range of 53.6-59.2mg/100g, indicate that the winter-spinach is a beneficial source of PCs.

Detection of flavonoids from Spinacia oleracea leaves using HPLC-ESI-QTOF-MS/MS and UPLC-QqQLIT-MS/MS techniques.[Pubmed:30259760]

Nat Prod Res. 2019 Aug;33(15):2253-2256.

Spinacia oleracea L. (Spinach) is a leafy vegetable which is considered to have a high nutritional value. Flavonoids in spinach were reported to act as antimutagenic property. Rapid detection of these flavonoids in Spinach was achieved by using HPLC-ESI-QTOF-MS/MS. Thirty six compounds were tentatively identified based on their retention times, accurate mass and MS/MS spectra. The fragmentation patterns of known compounds were applied to elucidate the structure of their corresponding derivatives having the same basic skeleton. Out of thirty six peaks, three peaks were assigned as patuletin and six peaks were assigned as Spinacetin derivatives. Twelve compounds were first time identified following the fragmentation pattern of known compounds. Five of the identified compounds i.e., Spinacetin, 5,3',4'-trihydroxy-3-methoxy-6,7-methylenedioxyflavone, protocatechuic acid, ferulic acid and coumaric acid were simultaneously quantified in spinach leaves by a validated UPLC-ESI-MS/MS method under MRM mode.

Anti-inflammatory Potential of Flavonoids from the Aerial Parts of Corispermum marschallii.[Pubmed:30109803]

J Nat Prod. 2018 Aug 24;81(8):1760-1768.

The isolation of phenolics from aerial parts of Corispermum marschallii yielded a total of 13 compounds including nine previously undescribed patuletin and Spinacetin glycosides. These were identified as patuletin 3- O-beta-d-galactopyranosyl-7- O-beta-d-glucopyranoside (1), Spinacetin 3- O-beta-d-galactopyranosyl-7- O-beta-d-glucopyranoside (2), patuletin 3- O-(6''- O-beta-d-glucopyranosyl)-beta-d-galactopyranoside (3), patuletin 3- O-(6''- O-alpha-l-arabinopyranosyl)-beta-d-galactopyranoside (4), patuletin 3- O-(2''- O-(5'''- O-alpha-l-arabinopyranosyl)-beta-d-apiofuranosyl)-beta-d-galactopyranoside (5), patuletin 3- O-(2''- O-beta-d-apiofuranosyl)-beta-d-galactopyranoside (6), Spinacetin 3- O-beta-d-galactopyranoside (7), patuletin 3- O-beta-d-galactopyranosyl-7- O-(6'''- O-feruloyl)-beta-d-glucopyranoside (8), and Spinacetin 3- O-beta-d-galactopyranosyl-7- O-(6'''- O-feruloyl)-beta-d-glucopyranoside (9). Structure elucidation was based on UV-visible, multistage MS, and 1D and 2D NMR spectroscopy and chemical derivatization, which allowed the identification on the glycosides with two different hexose moieties occurring at different positions of the aglycones. Most of the compounds tested inhibited the production of pro-inflammatory factors such as ROS, IL-8, and TNF-alpha in stimulated neutrophils.

Spinacetin Suppresses the Mast Cell Activation and Passive Cutaneous Anaphylaxis in Mouse Model.[Pubmed:30104977]

Front Pharmacol. 2018 Jul 30;9:824.

We previously reported the anti-inflammatory and anti-asthmatic activities of the extract of the Inula japonica Thunb. Aiming for discovery of a novel anti-inflammatory compound, we isolated Spinacetin from the extract and investigated its in vitro and in vivo anti-inflammatory effect and the related mechanism. Effect of Spinacetin on the Syk signaling pathway was studied in bone marrow-derived mast cells (BMMCs), and that on the nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein kinases (MAPKs) was investigated in Rat basophilic leukemia (RBL)-2H3 cells and human mast cell line (HMC-1). The in vivo anti-inflammatory activity was assessed with passive cutaneous anaphylaxis (PCA) reaction assay. Spinacetin significantly inhibited the release of histamine, and production of inflammatory mediators such as leukotriene C4 (LTC4) and interlukin-6 (IL-6) in IgE/Ag stimulated BMMCs. Analysis of the signaling pathways demonstrated that Spinacetin inhibited activation of Syk, linker of activated T cells (LAT), phospholipase Cgamma (PLCgamma), cytosolic phospholipase A2 (cPLA2), MAPKs, Akt/NF-kappaB, and intracellular Ca(2+) mobilization but with no effect on Fyn and Lyn. On the other hand, Spinacetin suppressed IgE/Ag-induced activation of RBL-2H3 cells with inhibition against phosphorylation of extracellular signal regulated-protein kinase (ERK), c-Jun-NH2-terminal kinase (JNK), p38 MAPKs, PLCgamma, translocation of cPLA2, and Akt/IkappaBalpha/NF-kappaB signal. However, Spinacetin had no effect on PMA and A23187-induced activation of HMC-1. Furthermore, oral administration of Spinacetin dose-dependently attenuated IgE/Ag-mediated PCA reaction in mouse model. Taken together, Spinacetin showed the activities in preventing inflammatory processes, which might be at least partially attributed to the abolishment of Syk-dependent activation of IgE/Ag-mediated mast cells.

Polyphenol Phase-II Metabolites are Detectable in Human Plasma after Ingestion of (13) C Labeled Spinach-a Pilot Intervention Trial in Young Healthy Adults.[Pubmed:29529352]

Mol Nutr Food Res. 2018 May;62(10):e1701003.

SCOPE: After intrinsic labeling of spinach (Spinacia oleracea L., Chenopodiaceae) with (13) CO2 , we investigated if labeled polyphenol metabolites were detectable in human plasma. METHODS AND RESULT: In a pilot intervention trial, five healthy men consumed 5 g freeze-dried (13) C labeled spinach, including a total amount of 160 mumol methoxyflavonols, including 70 mumol 5,3',4'-trihydroxy-3-methoxy-6,7-methylendioxyflavone-4'-glucuronide. Plasma samples of all subjects were analyzed with regard to their (13) C/(12) C ratio. Additionally, (13) C labeled metabolites of patuletin, Spinacetin, and 5,3',4'-trihydroxy-3-methoxy-6,7-methylendioxyflavone (TMM) were analyzed in plasma samples in a subgroup of three subjects. TMM-glucuronide, TMM-sulfate, and Spinacetin-glucuronide-sulfate, the latter as (12) C1(13) C16 and (13) C17 isotopologs, were tentatively identified. Plasma concentration of TMM-glucuronide and TMM-sulfate reached cmax from 19.1-54.3 and 22.5-125.5 nmol L(-1) , respectively, 7-9 h post-ingestion. CONCLUSION: It seems likely that (13) C labeled TMM-glucuronide and TMM-sulfate are phase-II metabolites which were converted after colonic transformation. Variations in plasma kinetics were observed for these two metabolites and may be attributed to the individual composition of the microbiota. We conclude that (13) C labeled polyphenol metabolites are detectable and quantifiable in human plasma.

Rapid ultra-high-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry and selected reaction monitoring strategy for the identification and quantification of minor spinacetin derivatives in spinach.[Pubmed:28836299]

Rapid Commun Mass Spectrom. 2017 Nov 15;31(21):1803-1812.

RATIONALE: Spinach is green leafy vegetable which is a rich source of flavonoids, phenolic acids, carotenoids, and vitamins A, C and E. It contains unique flavonoids which have significant anticarcinogenic, antiinflammatory and free radical scavenging activities. The present study reports the systematic identification and quantification of novel flavonoids by ultra-high-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry (UHPLC/HR-QTOFMS). METHODS: An ultrasonication technique was used for the extraction of flavonoids from spinach. A rapid and reliable analytical method was established for the identification of flavonoids from methanolic extract. Flavonoids were characterized by their ultraviolet (UV) spectra, high-resolution accurate masses and MS/MS fragmentation pathways obtained using electrospray ionization (ESI). Furthermore, precursor ions from the intact molecule, and the resulting product ions, were monitored by selected reaction monitoring (SRM) with different collision energies in positive and negative ion mode. RESULTS: For the first time, five minor Spinacetin derivatives were identified under optimized SRM and broadband collision-induced dissociation (+bbCID) conditions. Fragmentation pathways were proposed for spectra obtained in ESI positive ion mode. The use of HR-QTOFMS and SRM allowed us to differentiate between molecules with the same nominal mass. The identified Spinacetin derivatives were found to be acylated with ferulic and coumaric acids. CONCLUSIONS: UHPLC interfaced with HR-QTOFMS in combination with SRM provides a rapid, reliable and highly sensitive method for the identification of flavonoids, and potentially other bioactive compounds, in a complex matrix.

Chenopodium bonus-henricus L. - A source of hepatoprotective flavonoids.[Pubmed:28229939]

Fitoterapia. 2017 Apr;118:13-20.

Three new flavonoid glycosides (7-9) named patuletin-3-O-(5'''-capital O, Cyrillic-capital IE, Cyrillic-feruloyl)-beta-d-apiofuranosyl(1-->2)[beta-d-glucopyranosyl (1-->6)]-beta-d-glucopyranoside (7), Spinacetin-3-O-(5'''-capital O, Cyrillic-capital IE, Cyrillic-feruloyl)-beta-d-apiofuranosyl (1-->2)[beta-d-glucopyranosyl(1-->6)]-beta-d-glucopyranoside (8) and 6-methoxykaempferol-3-O-(5'''-capital O, Cyrillic-capital IE, Cyrillic-feruloyl)-beta-d-apiofuranosyl(1-->2)[beta-d-glucopyranosyl (1-->6)]-beta-d-glucopyranoside (9) together with six known flavonoid glycosides of patuletin, Spinacetin and 6-methoxykaempferol (1-6) were isolated from the aerial parts of C. bonus-henricus and identified with spectroscopic methods (1D and 2D NMR, UV, IR, HRESIMS). The MeOH extract exerts hepatoprotective and antioxidant activities comparable to those of flavonoid complex silymarin in in vitro (60mug/mL) and in vivo (100mg/kg/daily for 7days) models of hepatotoxicity, induced by CCl4. Flavonoids (1-9) (100muM), compared to silybin, significantly reduced the cellular damage caused by CCl4 in rat hepatocytes, preserved cell viability and GSH level, decreased LDH leakage and reduced lipid damage. High concentrations of compounds (1-9) showed marginal or no cytotoxicity on HepG2 cell line. The experiment data suggest that the glycosides of 6-methoxykaempferol, Spinacetin and patuletin are a promising and safe class of hepatoprotective agents.

6-Methoxyflavonol Glycosides with In Vitro Hepatoprotective Activity from Chenopodium bonus-henricus Roots.[Pubmed:26434121]

Nat Prod Commun. 2015 Aug;10(8):1377-80.

One new, namely 6-methoxykaempferol 3-O-[beta-apiofuranosyl(l-->2)]-f-glucopyranosyl(l->6)-fl-glucopyranoside (2), and two known flavonoid glycosides, Spinacetin 3-O-[beta-apiofuranosyl(1-->2)]-beta-glucopyranosyl(1-->6)-beta-glucopyranoside (1) and Spinacetin 3-O-gentiobioside (3), were isolated from the roots of Chenopodium bonus-henricus L. Their structures were determined by means of spectroscopic methods (ID, 2D NMR, UV, IR) and HR-ESI-MS. Radical scavenging and anti-oxidant activities of 1 and 3 were established using DPPH and ABTS free radicals, FRAP assay and inhibition of lipid peroxidation (LP) in a linoleic acid system by the ferric thiocyanate method. Compound 3 was found to possess stronger DPPH and ABTS radical scavenging activity (IC50 0.44 +/- 0.008 mM and 0.089 +/- 0.002 mM, respectively) compared with 1 (IC50 1.22 +/- 0.0 10 mM and 0.11 +/- 0.004 mM, respectively). Both flavonoids inhibited the lipid peroxidation of linoleic acid significantly. Additionally, 1 and 3 significantly reduced the cellular damage caused by the hepatotoxic agent CCI4 in rat hepatocytes and preserved cell viability and GSH level, decreased LDH leakage and reduced lipid damage. Effects were similar to those of the positive control silymarin. Control of self-toxic effects made in a MTT based assay using HepG2 cells revealed statistically significant cytotoxic effects only in very high concentrations (exceeding mM) and an incubation time of 72 h, making flavonoid glycosides with a 6-methoxykaempferol skeleton a promising and safe class of hepatoprotective compounds.

Phenolic Compounds from Atriplex littoralis and Their Radiation-Mitigating Activity.[Pubmed:26290401]

J Nat Prod. 2015 Sep 25;78(9):2198-204.

From the aerial parts of Atriplex littoralis, three new flavonoid glycosides named atriplexins I-III (1-3), a known flavonoid glycoside, Spinacetin 3-O-beta-d-glucopyranoside (4), arbutin (5), and 4-hydroxybenzyl-beta-d-glucopyranoside (6) were isolated. Their structures were elucidated on the basis of detailed spectroscopic analysis, including 1D and 2D NMR (COSY, NOESY, TOCSY, HSQC, HMBC) and HRESITOF MS data. The compounds were tested for in vitro protective effects on chromosome aberrations in peripheral human lymphocytes using a cytochalasin-B-blocked micronucleus (MN) assay in a concentration range of 0.8-7.4 muM of final culture solution. Chromosomal damage was induced by 2 Gy of gamma-radiation on binucleated human lymphocytes, and the effects of the compounds were tested 2 to 19 h after irradiation. The frequency of micronuclei (MNi) was scored in binucleated cells, and the nuclear proliferation index was calculated. The highest prevention of in vitro biochemical and cytogenetic damage of human lymphocytes induced by gamma-radiation was exhibited by 3 (reduction of MN frequency by 31%), followed by 4 and 6.

Topical analgesic, anti-inflammatory and antioxidant properties of Oxybaphus nyctagineus: phytochemical characterization of active fractions.[Pubmed:24945398]

J Ethnopharmacol. 2014 Aug 8;155(1):776-84.

ETHNOPHARMACOLOGICAL RELEVANCE: Oxybaphus nyctagineus (Michx.) Sweet has traditionally been used by several Native American tribes predominantly as a topical anti-inflammatory and analgesic agent. AIM OF THE STUDY: To evaluate the antioxidant, analgesic and anti-inflammatory activity of the extracts prepared from the aerial parts of Oxybaphus nyctagineus and to characterize the major chemical constituents of the bioactive extracts. MATERIALS AND METHODS: Crude polar and apolar extracts (PCE and ACE) of the herb of Oxybaphus nyctagineus were prepared and tested in the models of the CFA-induced hyperalgesia in rat knee and carrageenan-induced paw edema in rat. To identify the active compounds, subfractions were prepared by column chromatography and subjected in vitro assays, such as antioxidant assays (DPPH, peroxynitrite (ONOO-) scavenging), and the LPS-induced IL-1beta release test in human monocytes. Preparative HPLC was employed for the isolation of active substances, while phytochemical analysis was performed by mean of LC-MS/MS and NMR. RESULTS: The topically administered PCE and ACE of Oxybaphus nyctagineus demonstrated a significant analgesic and anti-inflammatory effect in the inflammation animal models. The subfraction A4 of ACE and the subfraction P5 of PCE considerably inhibited the LPS-induced IL-1beta release in human monocytes, while the strongest activity was localized in the subfraction P5 in the antioxidant assays. The HPLC-MS/MS and NMR analysis revealed that 6-methoxyflavonol diglycosides, namely patuletin-3-O-robinobioside (1), 6-methoxykaempferol-3-O-robinobioside (2), Spinacetin-3-O-robinobioside (3), and hydroxy-polyenoic fatty acids, namely corchorifatty acid B (4), 9-hydroxy-10E,12Z,15Z-octadecatrienoic acid (9-HOT acid) (5), and 9-hydroxy-10E,12Z-octadecadienoic acid (9-HOD acid) (6) were present in PCE, and in ACE as major compounds. CONCLUSION: The results of this study established a pharmacological evidence for the traditional use of Oxybaphus nyctagineus as an anti-inflammatory agent used topically, and provided data on its phytochemical composition for the first time.

[Anti-inflammatory constituents from Inula japonica].[Pubmed:24754174]

Zhongguo Zhong Yao Za Zhi. 2014 Jan;39(1):83-8.

Chemical constituents of Inula japonica were isolated and purified by repeated column chromatographies, over silica gel, and Toyopearl HW-40, and preparative HPLC. On the basis of spectral data analysis, including NMR and MS data, the structures of the isolates were elucidated and their anti-inflammatory activities were assayed. Fifteen compounds were isolated from the ethyl acetate extract of I. japonica, and their structures were elucidated as dihydrosyringenin (1), (3S, 5R, 6S, 7E)-5,6-epoxy-3-hydroxy-7-megastigmen-9-one (2), (6R, 7E) -9-hydroxy-4,7-megastigmadien-3-one (3), arnidiol (4), taraxasterol acetate (5), 8,9,10-trihydroxythymol (6), taxifolin (7), luteolin (8), napetin (9), eupatin (10), Spinacetin (11), quercetin (12), p-hydroxycinnamic acid (13), caffeic acid (14), and caffeoyl acetate (15). Compounds 1, 2, 7, 13 and 15 were isolated from the genus Inula for the first time, and compounds 3, 4, 9-11 and 14 were isolated from this plant for the first time. The anti-inflammatory activity result showed that compounds 3, 6-12 and 14 exhibited inhibition effect against leukotriene C4 (LTC4) synthesis and degranulation definitely in c-Kit Ligand (KL) induced mast cells, and compound 8 and 12 also had the suppression effect against lipopolysacharide(LPS) induced nitric oxide (NO) activity in RAW264.7 macrophages. It is firstly reported that compounds 7 and 9-11 possessed potent inhibition activities against LTC4 generation and degranulation in mast cells.

Spasmolytic activity of Artemisia copa aqueous extract and isolated compounds.[Pubmed:22577954]

Nat Prod Res. 2013;27(11):1007-11.

Artemisia copa Phil. (Compositae) is used in popular medicine as a digestive and for gastric pains. The effects of A. copa aqueous extract and its isolated compounds were evaluated on isolated rat jejunum. The extract inhibited non-competitively the cumulative concentration-response curves induced by acetylcholine and CaCl2. The tonic jejunum contractions induced by 80 mM KCl were inhibited by A. copa. Relaxant effects of A. copa on the tonic contraction induced by 25 mM KCl, [EC50: 0.94 mg mL(-1) (0.64-1.39)], was not inhibited by glibenclamide, TEA, l-NAME or methylene blue. Chrysoeriol, Spinacetin and luteolin (30 microg mL(-1)), produced an antagonism on the CaCl2 concentration-response curve, showing an inhibition of the maximum contractions (70.0% +/- 5.0%, 49.1% +/- 4.5% and 77.0% +/- 3.5% of E max, respectively), whereas tricin did not inhibit when the same concentration was used. A. copa exerts spasmolytic activity by blocking calcium channels and three isolated compounds could be, at least partly, responsible for the effect.

Flavonoids from Artemisia copa with anti-inflammatory activity.[Pubmed:16450301]

Planta Med. 2006 Jan;72(1):72-4.

Bioactivity-guided fractionation of the dichloromethane and ethanol extracts from the aerial parts of Artemisia copa led to the isolation of the flavonoids Spinacetin, jaceosidin, axillarin, penduletin, tricin and chrysoeriol. These compounds were studied for possible inhibitory activity on the generation of inflammatory mediators in a cell line of mouse macrophages (RAW 264.7) stimulated with lipopolysaccharide. Spinacetin and jaceosidin weakly inhibited nitric oxide production whereas all flavonoids reduced prostaglandin E2 levels to different extents. The most active flavonoid was jaceosidin that inhibited cyclooxygenase-2 activity in a concentration-dependent manner with an IC50 value of 2.8 microM. In addition, the other flavonoids partially inhibited synovial phospholipase A2 activity. These mechanisms may provide a basis for explaining the anti-inflammatory activity of this plant.