Chemical Profiling of Kaliziri Injection and Quantification of Six Caffeoyl Quinic Acids in Beagle Plasma by LC-MS/MS.

Vitiligo is a stubborn multifactorial skin disease with a prevalence of approximately 1% in the global population. Kaliziri, the seeds of Vernonia anthelmintica (L.) Willd., is a well-known traditional Uyghur medicine for the treatment of vitiligo. Kaliziri injections is a Chinese-marketed treatment approved by the China Food and Drug Administration for the treatment of vitiligo. The significant effects of Kaliziri injection have been thoroughly studied. However, chemical components studies and plasma quantification studies are lacking for Kaliziri injection. Ultra-high-performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry was employed to comprehensively characterize the caffeoyl quinic acid derivatives present in Kaliziri injection. Based on accurate mass measurements, key fragmental ions and comparisons with reference standards, 60 caffeoyl quinic acid derivatives were identified in Kaliziri injections, including caffeoyl quinic acids, coumaroyl caffeoyl quinic acids, dicaffeoyl quinic acids, feruloyl caffeoyl quinic acids, and dicaffeoyl quinic acid hexosides. Moreover, an HPLC-MS/MS method was developed and validated for the quantitative analysis of 5-caffeoyl quinic acid, 4-caffeoyl quinic acid, 1,3-dicaffeoyl quinic acid, 3,4-dicaffeoyl quinic acid, 3,5-dicaffeoyl quinic acid and 4,5-dicaffeoyl quinic acid in beagle plasma. The quantitative HPLC-MS/MS method was applied to quantify these six major caffeoyl quinic acids in beagle plasma after the subcutaneous administration of Kaliziri injection. All of the six analytes reached their peak plasma of concentrations within 30 min.

Anti-nociceptive and Anti-inflammatory Properties of Ilex latifolia and its Active Component, 3,5-Di-caffeoyl Quinic Acid Methyl Ester

The present study was conducted to investigate anti-nociceptive and anti-inflammatory effects of the leaves of Ilex latifolia Thunb (I. latifolia) in in vivo and in vitro. Writhing responses induced by acetic acid and formalin- and thermal stimuli (tail flick and hot plate tests)-induced pain responses for nociception were evaluated in mice. I. latifolia (50 – 200 mg/kg, p.o.) and ibuprofen (100 mg/kg, p.o.), a positive non-steroidal anti-inflammatory drug (NSAID), inhibited the acetic acid-induced writhing response and the second phase response (peripheral inflammatory response) in the formalin test, but did not protect against thermal nociception and the first phase response (central response) in the formalin test. These results show that I. latifolia has a significant anti-nociceptive effect that appears to be peripheral, but not central. Additionally, I. latifolia (50 and 100 µg/mL) and 3,5-di-caffeoyl quinic acid methyl ester (5 µM) isolated from I. latifolia as an active compound significantly inhibited LPS-induced NO production and mRNA expression of the pro-inflammatory mediators, iNOS and COX-2, and the pro-inflammatory cytokines, IL-6 and IL-1β, in RAW 264.7 macrophages. These results suggest that I. latifolia can produce antinociceptive effects peripherally, but not centrally, via anti-inflammatory activity and supports a possible use of I. latifolia to treat pain and inflammation.

Caffeoyl quinic acids in Chrysanthemum morifolium improve LPS-induced HUVEC vascular endothelial cell injury by regulating ERK/MAPK signaling pathway / 药学学报

To explore the effect of total extract of Chrysanthemum morifolium on lipopolysaccharide (LPS)-induced acute lung injury in mice, we studied the effects of three caffeoyl quinic acids isolated from Chrysanthemum morifolium on vascular endothelial cell injury and their mechanisms of action. All animal experiments were carried out strictly in accordance with the National Animal Welfare Ethics and Protection Regulations. A mouse model of acute lung injury was established by intranasal instillation of LPS. After 6 days of oral administration of chrysanthemum extract, the lung wet weight/dry weight ratio, tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) were measured in mouse bronchoalveolar lavage fluid. Human umbilical vein endothelial cells (HUVEC) were serum starved for 12 h and treated with 2.5 μg·mL-1 LPS for 24 h to establish the in vitro model of vascular endothelial cell injury. After 24 h of treatment of caffeoyl quinic acids from Chrysanthemum morifolium, the levels of TNF-α, IL-6, IL-1β, vascular cell adhesion molecule-1 (VCAM-1) and endothelin-1 (ET-1) were measured by ELISA in the cell culture supernatant, the malondialdehyde (MDA) level was detected by TBA method, the superoxide dismutase (SOD) level was determined by hydroxylamine method, and the nitric oxide (NO) level was assayed by a one-step method. The levels of p-MEK1/2, MEK1/2, p-ERK1/2, ERK1/2, p-JNK, JNK, p-P38 and P38 of mitogen-activated protein kinase (MAPK) signaling pathway were detected by Western blot. The total extract of Chrysanthemum morifolium can significantly reduce the wet weight/dry weight ratio of lung in mice and the levels of TNF-α, IL-6 and IL-1β in alveolar lavage fluid. The caffeoyl quinic acids from Chrysanthemum morifolium significantly increased the levels of SOD and NO, decreased the levels of TNF-α, IL-6, IL-1β, VCAM-1, ET-1 and MDA, and significantly reduced the levels of p-MEK1/2, p-ERK1/2. In conclusion, total extracts of Chrysanthemum morifolium exhibit certain protective effect on mice with acute lung injury, and three caffeoyl quinic acids from Chrysanthemum morifolium may improve LPS-induced vascular endothelial cell injury by inhibiting inflammatory cytokines and oxidative stress, and regulating inter-cellular adhesion molecule and vasomotor factors through ERK/MAPK signaling pathway.

Anti-oxidant chemical constituents from Patrinia villosa / 中草药

Objective: To study the antioxidant chemical constituents and antioxidant activity of Patrinia villosa. Methods: The 70% ethanol-water extract of the herb was separated by silica gel column chromatography, ODS column chromatography and sephadex column chromatography. Then, the compound were further purified and extracted by semi-preparative HPLC. Their structures were elucidated by physiochemical property and spectral analysis. DPPH and ABTS methods were used to determine the antioxidant bioactivities of the isolated compounds. Results: A total of ten compounds were isolated and synthesized, including chlorogenic acid butyl ester (1), 3,4-di-O-caffeoyl quinic acid methyl ester (2), luteolin-7-O-rutinoside (3), 1β-O-β-D-glucopyranosy- 15-O-(p-hydroxylphenylacetate)-5α,6βH-eudesma-3,11(13)-dien-12,6α-olide (4), 3,4-di-O-caffeoyl quinic acid ethyl ester (5), 4,5-di-O-caffeoyl quinic acid methyl ester (6), 4,5-di-O-caffeoyl quinic acid n-butyl ester (7), luteolin-7-O-β-D-glucuronide methyl ester (8), luteolin-7-O-β-D-glucuronide ethyl ester (9), and apigenin-7-O-β-D-glucuronide methyl ester (10). The DPPH radical scavenging IC50 of compounds 3, 8, and 9 were (23.95 ± 0.71), (73.09 ± 0.33), and (25.06 ± 0.65) μmol/L, respectively. The ABTS radical scavenging IC50 was (7.13 ± 0.07), (11.48 ± 0.21), (5.15 ± 0.08) mol/L, respectively. Conclusion: Eight compounds except compounds 3 and 8 are obtained from this species for the first time. Compounds 3, 8, and 9 had significant antioxidant activity.

Chemical constituents from stems of Acanthopanax henryi / 中草药

Objective To study the chemical constituents from stems of Acanthopanax henryi based on LPS-induced macrophages RAW264.7 and microglia BV2 as the bioactivity guided model. Methods The compounds were isolated and purified by silica gel and Sephadex LH-20 column chromatography, as well as Prep-TLC and recrystallization methods. Their structures were identified on the basis of their physicochemical properties and spectroscopic data. Results Eighteen compounds were obtained from A. henryi and their chemical structures were identified as p-hydroxybenzoic acid (1), trans-p-hydroxycinnamic acid (2), (E)-caffeic acid methyl ester (3), caffeic acid (4), trans-coniferyl aldehyde (5), syringaldehyde (6), vanillin (7), 6-methoxy-7-hydroxycoumarin (8), trans-sinapaldehyde (9), undecane-1,11-dioic acid monomethyl ester (10), (-)-sesamin (11), 3-O-caffeoyl-quinic acid (12), 5-O-caffeoyl-quinic acid (13), 1,3-di-O-caffeoyl-quinic acid (14), 1,4-di-O-caffeoyl-quinic acid (15), 1,5-di-O-caffeoyl-quinic acid (16), stigmasterol (17), and β-sitosterol (18), respectively. Conclusion To the best of our knowledge, compound 10 was isolated from Araliaceae for the first time. Except compounds 12, 14, 17, and 18, all of other compounds were obtained from this species for the first time.

Urine metabolomics shows an induction of fatty acids metabolism in healthy adult volunteers after supplementation with green coffee (Coffea robusta L.) bean extract.

BACKGROUND AND OBJECTIVE: Green coffee bean extract is used as herbal medicine or supplement for weight reduction and obesity. The active constituents are considered caffeine and chlorogenic acid (CGA) derivatives. The mode of action of CGA is still unclear and can be related to peroxisome proliferator-activated receptor α (PPAR-α) and liver X receptor Rα (LXR-α). Metabolomics may be an innovative tool for the description and discovery of the multiple target nature of such phytocomplex. METHODS: 24 h urine samples were collected once a week from ten healthy adult volunteers consuming daily 400 mg of dry Green coffee bean extract (GCBE, 4.9% of chlorogenic acid) each day for 30 days (5 harvesting days, considering also the first day of supplementation). Urine samples were analyzed by LC-QTOF using both untargeted and targeted approaches. The latter was used to monitor two urinary markers of oxidative stress (allantoin, 8-OHdG). RESULTS: Metabolomics analysis (PLS-DA) revealed changes in urine composition before and during the treatment with GCBE. Markers related to treatment were metabolites related to polyphenol administration as hippuric acid, benzoic acid derivatives, dihydroferulic and dihydrosinapic acid sulphate, but also carnitine derivatives and dicarboxylic acids. On the other hand, no changes in the levels of allantoin and 8-OHdG were observed. CONCLUSION: This preliminary study showed the possible usefulness of metabolomics approach in the evaluation of GCBE consumption in healthy subjects. The observed changes in urinary composition can be related to the catabolism of GCBE constituents and to induced fatty acid metabolism, mainly related to carnitine derivatives. This latter result could be considered, at least in part, as a further proof of the mode of action of green coffee extract.

Selective enzymatic hydrolysis of chlorogenic acid lactones in a model system and in a coffee extract. Application to reduction of coffee bitterness.

Chlorogenic acid lactones have been identified as key contributors to coffee bitterness. These compounds are formed during roasting by dehydration and cyclization of their precursors, the chlorogenic acids (CGAs). In the present study, we investigated an approach to decompose these lactones in a selective way without affecting the positive coffee attributes developed during roasting. A model system composed of (3-caffeoylquinic acid lactone (3-CQAL), 4- caffeoyl quinic acid lactone (4-CQAL), and 4-feruloylquinic acid lactone (4-FQAL)) was used for the screening of enzymes before treatment of the coffee extracts. Hog liver esterase (HLE) hydrolyzed chlorogenic acid lactones (CQALs, FQALs) selectively, while chlorogenate esterase hydrolyzed all chlorogenic acids (CQAs, FQAs) and their corresponding lactones (CQALs, FQALs) in a non-selective way. Enzymatically treated coffee samples were evaluated for their bitterness by a trained sensory panel and were found significantly less bitter than the untreated samples.

Caffeoylquinic acids from antiplasmodial active extract of Xanthium cavanillesii fruits and their molecular modelling studies.

The antiplasmodial active extract of Xanthium cavanillesii contains 3,4-dicaffeoyl quinic acid (3,4-DCQA), 3,5-dicaffeoyl quinic acid (3,5-DCQA) and 1,3,5-tricaffeoyl quinic acid (1,3,5-TCQA). These results inspired us to investigate the interaction of these molecules with a promising validated target of Plasmodium, PfATP6 orthologue of mammalian Ca+2-ATPase. Models of this receptor were obtained through comparative modelling. Afterwards, molecular docking studies were used to identify possible interaction modes of these caffeoyl quinic derivatives on the binding site. The 1,3,5-TCQA had the best energy, but all of these had better energy than thapsigargin, a non-competitive inhibitor of the sarco/endoplasmatic reticulum Ca+2-ATPase (SERCA).

Analysis on chemical composition of Farfarae Flos by UPLC-Q-TOF-MS / 中草药

Objective: The chemical constituents in the methanol extract from Farfarae Flos were rapidly identified using UPLC-Q-TOF-MS in positive and negative ion modes. Methods: The analysis was performed on an Agilent Poroshell 120 EC-C18 chromatographic column (100 mm × 2.1 mm, 2.7 μm). The mobile phase consisted of acetonitrile and 0.1% aq formic acid. In positive ion mode, gradient elution: 0-1 min, 5%-17% B; 1-3 min, 17%-19% B; 3-14 min, 19%-44% B; 14-16 min, 44%-66% B; 16-26 min, 66%-87% B; 26-28 min, 87%-95% B; 28-33 min, 95% B. In negative ion mode, gradient elution: 0-2 min, 5%-14% B; 2-10 min, 14%-32% B; 10-15 min, 32% B. The flow rate was 0.4 mL/min, and the injection volume was 5 μL. The information of the compounds was analyzed by positive and negative ion modes mass spectrum information, elements composition, reference substance retention time or mass spectrum parameters of compounds in literature. Results: Thirty-four compounds in Farfarae Flos extracts were identified, combined with provided accurate molecular weight compounds by UPLC-Q-TOF-MS, including 12 kinds of terpenoids, eight kinds of flavonoids, seven kinds of phenolic acids, two kinds of pyran compounds, one kind of phenolic ketones, one kind of fat ketones, and three kinds of alkaloids. Conclusion: The method provides the theory basis for quality control and the clinical reasonable application and provides the reference for clarifying its efficacy material base.

Phenolic constituents from Erycibe schmidtii / 中草药

Objective: To investigate the chemical constituents of Erycibe schmidtii. Methods: The compounds were separated and purified by silica gel and Sephadex LH-20 column chromatography, and semipreparative HPLC. Their structures were determined by nuclear magnetic resonance (NMR) spectroscopy analysis method. The anti-inflammatory activity of some compounds was tested by relative NO productivity. Results: Twelve compounds were isolated and identified as caffeic acid (1), N-trans-p-hydroxyphenethyl ferolamine (2), chlorogenic acid (3), chlorogenic acid methyl ester (4), 4-O-caffeoylquinic acid (5), 4-O-caffeoylquinic acid methyl ester (6), 4,5-di-O-caffeoylquinic acid (7), 4,5-di-O-caffeoylquinic acid methyl ester (8), 3,5-di-O-caffeoylquinic acid (9), 3,5-di-O-caffeoylquinic acid methyl ester (10), 3,4-di-O-caffeoylquinic acid (11), and 3,4-di-O-caffeoylquinic acid methyl ester (12). Compounds 1 and 2 showed some inhibitory activity for RAW cells 264.7 releasing NO. Conclusion: Compounds 5-7, 9, 11, and 12 are obtained from the plants of Erycibe Roxb. for the first time, while compounds 1, 2, and 4-12 are separated from this plant for the first time. Compounds 1 and 2 exhibit certain anti-inflammatory activity.

Chemical constituents in stems of Ilex cornuta / 中草药

Objective: To investigate the chemical constituents in the stems of Ilex cornuta and the ability of scavenging free radicals of compounds 1-9. Methods: The compounds were isolated and purified by silica gel, medium pressure column chromatography, and semi-preparative liquid chromatography, and their structures were elucidated by chemical properties and spectroscopic analyses. The antifreeradical efficiency of compounds 1-9 was evaluated by DPPH radical scavenging assay. Results: Fifteen compounds were isolated and their structures were identified as isochlorogenic acid B (1), 3,4,5-tricaffeoylquinic acid (2), 4,5-di-O-caffeoyl quinic acid methyl ester (3), 3,4-di-O-caffeoyl quinicacid methyl ester (4), 3,5-di-O-caffeoyl quinicacid methyl ester (5), 3,4,5-tri-O-caffeoyl quinic acid methyl ester (6), 3,5-dimethoxy-4-hydroxybenzaldehyde (7), ethyl gallate (8), dihydrosyringenin (9), 2,6-dimethoxy-1,4-benzoquinone (10), arctigenin (11), 1-O-(vanillic acid)-6-O-(3″, 5″-dimethoxy-galloyl)-β-D-glycoside (12), (+)-1-hydroxypinoresinol-1-O-β-D-glucopyranoside (13), (+)-(7S,8S)-syringylglycerol 8-O-β-D-glucopyranoside (14), and schaftoside (15). Compounds 1-7 had good antifreeradical efficiency. Conclusion: Compounds 6,8-10,14, and 15 are obtained from the plants of Ilex L. the first time, and compounds 2,7,11, and 12 are obtained from this plant for the first time. Compounds 1-6 have good antifreeradical efficiency.

High performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight-mass spectrometry as a powerful analytical strategy for systematic analysis and improved characterization of the major bioactive constituents from Radix Dipsaci.

Radix Dipsaci (RD), the dried root of Dipsacus asper, is commonly used as a traditional Chinese medicine for the treatment of bone diseases and functions in strengthening bone and healing bone fractures. Nevertheless, the high polarity, non chromophores and low abundance of multiple compounds in this plant bring difficulty for their isolation and structural determination by traditional chromatographic and spectroscopic techniques, which hindered the use of RD in clinical practice and retarded the process of RD modernization. In this work, a sensitive and rapid high-performance liquid chromatography coupled with electrospray time-of-flight-mass spectrometry (HPLC-ESI-QTOF-MS/MS) was employed to rapidly separate and identify the multiple minor constituents in RD. Separation was performed an Agilent poroshell 120 EC-C18 column (2.1mm×100mm, i.d., 2.7µm) with 0.1% formic acid aqueous solution and acetonitrile as the mobile phase under gradient conditions. As a result, 36 major constituents including dipsacus saponins, iridoid glycosides and caffeoyl quinic acid derivatives were identified or tentatively characterized from the RD, 11 of which had not been previously reported to the best of our knowledge. In conclusion, the HPLC-ESI-QTOF-MS/MS is feasible and credible technique to separate and identify the constituents in complex matrices of traditional Chinese medicines.

Anti-Aspergillus activity of green coffee 5-O-caffeoyl quinic acid and its alkyl esters.

The antifungal activities of 5-O-caffeoyl quinic acid (5-CQA) and of methyl, butyl, octyl, and dodecyl esters or 5-CQA, were tested on five toxigenic moulds from the Aspergillus genus (Aspergillus flavus, Aspergillus nomius, Aspergillus ochraceus, Aspergillus parasiticus, Aspergillus westerdijkiae). These mycotoxin producers' moulds may contaminate many types of food crops throughout the food chain posing serious health hazard to animals and humans. The use of chemical methods to decrease mycotoxin producer moulds contamination on food crops in the field, during storage, and/or during processing, has been proved to be efficient. In this work, the antifungal effect of 5-CQA and a homologous series of 5-CQA esters (methyl, butyl, octyl, dodecyl), was investigated using the microdilution method and the minimum inhibitory concentrations (MIC50 and MIC80). All molecules presented antifungal activity, and two esters showed a MIC for all fungi: octyl (MIC50 ≤ 0.5-0.75 mg/mL, MIC80 = 1.0-1.5 mg/mL) and dodecyl (MIC50 = 0.75-1.25 mg/mL) chlorogenates. Dodecyl chlorogenate showed a MIC80 (1.5 mg/mL) only for A. parasiticus. The maximum percent of growth inhibition on aspergillii was observed with octyl (78.4-92.7%) and dodecyl (54.5-83.7%) chlorogenates, being octyl chlorogenate the most potent antifungal agent. It was thus concluded that lipophilization improved the antifungal properties of 5-CQA, which increased with the ester alkyl chain length, exhibiting a cut-off effect at 8 carbons. As far as we know, it is the first report demonstrating that lipophilization may improve the antifungal activity of 5-CQA on five toxigenic moulds from the Aspergillus genus. Lipophilization would be a novel way to synthesize a new kind of antifungal agents with a good therapeutic value or a potential use as preservative in food or cosmetics.

Caffeoylquinic acid derivatives from Bidens parviflora and their antihistamine release activites / 中草药

Objective Based on the activities of antihistamine release to study the compounds from Bidens parvi-flora and find biological active compounds. Methods The chemical constituents from B.parviflora were isolated by silica gel and Sphadex LH-20 column chromatographies and purified by preparative HPLC. The chemical structures had been identified by physiochemical properties and spectroscopic methods. Results Six caffeoylquinic acid derivatives were identified as 3, 5-di-O-caffeoylquinic acid ( Ⅰ ),3, 4-di-O-caffeoylquinic acid ( Ⅱ ), 4, 5-di-O-caffeoylquinic acid ( Ⅲ ), 4-O-caffeoylquinic acid ( Ⅳ ), 5-O-caffeoylquinic acid ( Ⅴ ), 4-[3-(3, 4-dihydroxy-phenyl)-acryloyloxy]-2, 3-dihydroxy-2-methyl-butyric acid ( Ⅵ ). Conclusion Compounds Ⅰ - Ⅵ are first obtained from B. parviflora and Ⅵ is new one. Some of the compounds exhibit the activities in antiallergic assays. Moreover, the structure-activity relationships of these compounds have been also discussed in this paper.