Screening and mechanism study of components targeting DNA from the Chinese herb <i>Lonicera japonica</i> by liquid chromatography/mass spectrometry and fluorescence spectroscopy

Reverse phase-solid phase extraction from Cassia alata leaves (CaRP) was used to obtain a refined extract. Higher than wild-type sensitivity to CaRP was exhibited by 16 haploid Saccharomyces cerevisiae mutants with defects in DNA repair and membrane transport. CaRP had a strong DPPH free radical scavenging activity with an IC50 value of 2.27 μg mL−1 and showed no pro-oxidant activity in yeast. CaRP compounds were separated by HPLC and the three major components were shown to bind to DNA in vitro. The major HPLC peak was identified as kampferol-3-O-β-d-glucoside (astragalin), which showed high affinity to DNA as seen by HPLC-UV measurement after using centrifugal ultrafiltration of astragalin-DNA mixtures. Astragalin-DNA interaction was further studied by spectroscopic methods and its interaction with DNA was evaluated using solid-state FTIR. These and computational (in silico) docking studies revealed that astragalin-DNA binding occurs through interaction with G-C base pairs, possibly by intercalation stabilized by H-bond formation.

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“…For this, centrifugal ultrafiltration sampling followed by HPLC analysis was used, a fast screening for bioactive compounds binding to DNA [19]. …”

Section: Resultsmentioning

confidence: 99%

“…Normally, small molecules have to some extent selectivity except for their binding to DNA by electrostatic interaction (along the external DNA double helix) [19]. Intercalative binding is usually affected by planarity of the bound candidates and groove binding is related to the two types of grooves in DNA, i.e.…”

Section: Resultsmentioning

confidence: 99%

Astragalin from Cassia alata Induces DNA Adducts in Vitro and Repairable DNA Damage in the Yeast Saccharomyces cerevisiae

Saito

Silva

Santos

et al. 2012

IJMS

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“…Gradient elution with the content of the organic component varying over broad values was used to analyze mixtures of compounds with highly different polarities (plant pharmaceutical preparations, food products) or to determine different metabolites [27]. The HPLC analysis time could reach 120 min [29].…”

Section: Analytical Methods For Biological Samples From Pk Studies Ofmentioning

confidence: 99%

“…SPE is often used as a secondary and additional purification step, e.g., after preliminary ultrafiltration [27]. Three isoflavonoid glycosides were determined in rat plasma after oral administration of Astragalus mongholicus extract [28].…”

Section: Analytical Methods For Biological Samples From Pk Studies Ofmentioning

confidence: 99%

Pharmacokinetics of Quercetin and Other Flavonols Studied by Liquid Chromatography and LC-MS (a Review)

Guglya

2014

Pharm Chem J

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Modern analytical methods used to study the pharmacokinetics of quercetin and other flavonols, i.e., biologically active compounds that exhibit various therapeutic properties, were reviewed. The preparation of biological fluids and tissues for analysis, chromatography conditions, and mass spectrometric detection for various flavonols as the aglycons and glycosides and their metabolites were discussed. Quantitative analyses of flavonol concentrations in biological samples and their mass spectrometric identification in in vitro and in vivo studies during tests with pure compounds and multi-constituent plant extracts were presented as typical examples.Keywords: quercetin, flavonols, flavonoids, flavonoid pharmacokinetics, bioanalytical methods, liquid chromatography-mass spectrometry (LC-MS/MS).Flavonols are a subclass of flavonoids, a varied and broad class of polyphenolic compounds that includes the flavonol quercetin, one of the most widely distributed and studied representatives. These compounds were well known over 100 years ago as plant pigments. However, the discovery in the 1990s of the antioxidant properties of flavonoids stimulated renewed interest in them. They are capable of neutralizing free radicals, which is responsible for their various therapeutic properties [1,2].A pharmacokinetic (PK) study in vivo in humans and experimental animals, i.e., absorption, distribution, metabolism, and elimination, is a necessary component of the evaluation of the mechanisms of possible therapeutic or toxic action of a biologically active compound. The PK parameters of drug candidates are related to the effectiveness and administration mode in order to establish the optimum concentrations.The properties of quercetin and other flavonols were briefly characterized and the development in the last decade of methods for determining the concentrations and identifying flavonols in biological media during PK studies were discussed in the review. Structure, properties and natural sources of flavonolsFlavonoids are secondary plant metabolites and share common biosynthetic pathways. They are responsible for the color of fruits and leaves. Surface tissues of plants are especially rich in them [3]. They are involved in the photosynthesis, proliferation, and death (through apoptosis) of plant cells. It is assumed that the main biological role of flavonoids is to protect plants from aggressive environmental factors (UV radiation, infectious agents).The variety of flavonoids is explained by the fact that most of them occur in plants as O-or C-glycosides. The aglycon is a 15-carbon skeleton, the principal elements of which are two aromatic rings joined by a pyran ring. The last determines to which subclass one flavonoid or another belongs. The classification includes the subclasses flavones, flavonols, flavanones (catechins), anthocyanidins, isoflavones, and flavanonols [4].

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“…Thus, CU became a useful technique for screening active compounds bound to biomacromolecules such as bovine serum albumin, 5 α-glucosidase, 6 quinone reductase-2, 7 deoxyribonucleic acid (DNA) and liposomes. 8,9 High performance liquid chromatography-mass spectrometry (HPLC-MS or LC-MS) has been widely applied for the simultaneous separation and identification of active compounds in complex mixtures. 10 The combination of CU and LC-MS (CU-LC-MS) became a powerful tool in analyzing active compounds due to its simple operation, high speed and low sample consumption.…”

Section: Introductionmentioning

confidence: 99%

Analysis of Xanthine Oxidase Inhibitors fromPuerariae flosUsing Centrifugal Ultrafiltration Coupled with HPLC-MS

Liu¹,

Xiao²,

Ma³

et al. 2016

Journal of the Brazilian Chemical Society

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In this study, centrifugal ultrafiltration coupled with high performance liquid chromatographymass spectrometry was utilized to screen and identify xanthine oxidase inhibitors from Puerariae flos extract. The experimental conditions of centrifugal ultrafiltration including xanthine oxidase concentration, incubation time, pH and temperature were optimized. At the optimum condition (xanthine oxidase concentration: 30.0 μg mL -1 , incubation time: 20 min, pH 7.0 and temperature: 25 °C), four compounds were successfully screened from P. flos extract and identified as tectoridin, daidzin, ononin and biochanin A. The yields of tectoridin, daidzin, ononin and biochanin A were 0.231, 0.117, 0.303 and 0.089 g from 50.0 g crude P. flos samples. The inhibitory activities of these compounds were verified by xanthine oxidase inhibition assays. The experimental half maximal inhibitory concentration (IC 50 ) values of tectoridin, daidzin, ononin and biochanin A were 88.5, 85.1, 88.8 and 87.0 μmol L -1 , and the binding degree of them were 5. 70, 8.28, 6.31 and 37.83% at the optimum condition, respectively. The proposed method provided a rapid and effective way to screen and analyze active compounds from natural products. Keywords: HPLC-MS, inhibitor, Puerariae flos, ultrafiltration, xanthine oxidase IntroductionNatural products have been used as the most consistently successful resources of new drug discovery for a long time because of their great diversity of the chemical structures and better drug-like properties compared to the synthetic compounds.1 However, natural products resources like plant extracts were very complex and usually contained various kinds of components. Separation and purification of natural compounds were time-consuming and laborious processes.2 Hence, simple and effective methods aiming at directly screening natural product extracts would be greatly helpful for drug discovery. Centrifugal ultrafiltration (CU) utilized centrifugal force and a semi-permeable membrane to retain suspended solids and high molecular weight solutes, while liquid and low molecular weight solutes were allowed to pass through depending on the nominal molecular weight cut-off of the membrane. 4 Based on these features, active compounds could be retained by membrane together with enzyme due to the binding with enzyme. Thus, CU became a useful technique for screening active compounds bound to biomacromolecules such as bovine serum albumin, 5 α-glucosidase, 6 quinone reductase-2, 7 deoxyribonucleic acid (DNA) and liposomes. 8,9 High performance liquid chromatography-mass spectrometry (HPLC-MS or LC-MS) has been widely applied for the simultaneous separation and identification of active compounds in complex mixtures. 10The combination of CU and LC-MS (CU-LC-MS) became a powerful tool in analyzing active compounds due to its simple operation, high speed and low sample consumption. Compared with immobilized enzyme screening assay, complex synthesis procedures could be avoided in CU-LC-MS as well.11 Moreover, the efficient screeni...

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Screening and mechanism study of components targeting DNA from the Chinese herb Lonicera japonica by liquid chromatography/mass spectrometry and fluorescence spectroscopy

Cited by 24 publications

References 30 publications

Astragalin from Cassia alata Induces DNA Adducts in Vitro and Repairable DNA Damage in the Yeast Saccharomyces cerevisiae

Astragalin from Cassia alata Induces DNA Adducts in Vitro and Repairable DNA Damage in the Yeast Saccharomyces cerevisiae

Pharmacokinetics of Quercetin and Other Flavonols Studied by Liquid Chromatography and LC-MS (a Review)

Analysis of Xanthine Oxidase Inhibitors fromPuerariae flosUsing Centrifugal Ultrafiltration Coupled with HPLC-MS

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