High-performance liquid chromatography and LC-ESI-MS method for the identification and quantification of two biologically active isomeric coumarinolignoids cleomiscosin A and cleomiscosin B in different extracts of Cleome viscosa

A rapid, sensitive and simple reverse-phase high-performance liquid chromatographic-electrospray ionization-mass spectrometry method for simultaneous determination of cleomiscosin A and cleomiscosin B has been developed and validated. The isomeric coumarinolignoids cleomiscosin A (1) and cleomiscosin B (2) were separated on a Waters symmetry C(18) column with a solvent system composed of acetonitrile-methanol (1:2) and acetic acid-water (0.5:99.5) in a gradient elution mode. The absorption at 326 nm was chosen as the measuring wavelength in which resolution and baseline separation of compounds 1 and 2 could be obtained. The identity of the two isomeric compounds 1 and 2 in the samples were determined on a triple quadrupole mass spectrometer with ESI interface operating in the positive mode. Calibration curves were linear (r(2) > 0.993) over the concentration range 20-200 microg/mL for cleomiscosin A and 10-200 microg/mL for cleomiscosin B with acceptable accuracy and precision, respectively. The intra-day and inter-day precision were 1.13 and 0.82% for cleomiscosin A and 1.78 and 1.28% for cleomiscosin B, respectively. The validated method was successfully applied for the analysis of the above two compounds in different extracts of Cleome viscosa.     

A methoxylated quercetin glycoside harnesses HCC tumor progression in a TP53/miR-15/miR-16 dependent manner

Abstract

This study focused on studying the impact of flavonoids isolated from Cleome droserifolia on HCC cell lines and to further unveil their possible impact on TP53 and its downstream tumor suppressor miRNAs. Three flavonol glycosides were isolated from C. droserifolia namely, Isorhamnetin-3-O-β-D-glucoside (1), Quercetin-3`-methoxy-3-O-(4``-acetylrhamnoside)-7-O-α-rhamnoside (2), and Kaempferol-4`-methoxy-3,7-O-dirhamnoside (3). They showed a concentration and time dependent reduction in cellular viability and anchorage-independent growth of HCC cells. Moreover, they exhibited a decrease in the migrating capacity of HepG2 cells in a pattern similar to positive control cells. (2) Showed the most potent effects in halting HCC tumorigenic activity (IC₅₀=36?±?1.70?µM) and a repression of the cellular proliferation rate of HepG2 cells. Restoration of TP53 and its downstream tumor suppressor miRNAs; miR-15a, miR-16, miR-34a by (2) was observed. Moreover, attenuation of (2) mediated actions was shown upon using anti-miR-15a and anti-miR-16. To conclude, this study crystallizes a novel role of C. droserifolia in harnessing HCC progression in-vitro with a possible contribution of TP53/miR-15a/miR-16.     

A New Indole Alkaloid from Cleome droserifolia

The phytochemical investigations on Cleome droserifolia resulted in the isolation and characterization of a new indole alkaloid, 5‐hydroxy‐2‐methoxy‐1‐methyl‐1H‐indole‐3‐carbaldehyde ( 1 ). The structure elucidation was carried out on the basis of 1D‐ and 2D‐NMR techniques. In addition to 1 , two known aromatic derivatives, veratrol ( 2 ) and 2‐methoxy‐4‐methylacetophenone ( 3 ), were also obtained. All these compounds were purified by repeated column chromatography of the CH₂Cl₂ fraction obtained from MeOH extract of Cleome droserifolia. The structure of the new compound 1 was finally confirmed by the combined 1D (¹H‐ and ¹³C‐) and 2D (H? C correlations; HMBC and HSQC) NMR and IR spectroscopy, mass spectrometry (MS), and UV absorption spectroscopy techniques. The comparison of the physical and spectroscopic data with those in the literature provided evidence for the structure confirmation of known compounds. All the purified compounds were subjected to urease and α‐glucosidase enzymes inhibition. The results showed that compound 1 was more potent with an IC₅₀ value 11.97±2.067 μg/ml towards urease inhibition, while the activity of α‐glucosidase enzyme was marginal.     

Identification and quantification of two antihepatotoxic coumarinolignoids cleomiscosin A and cleomiscosin B in the seeds of Cleome viscosa using liquid chromatography-tandem mass spectrometry

A sensitive liquid chromatography/electrospray ionization tandem mass spectrometric (LC/ESI-MS/MS) method was developed for the identification and quantification of two antihepatotoxic coumarinolignoids cleomiscosin A and cleomiscosin B in different extracts of the seeds of Cleome viscosa. The separation of cleomiscosin A and cleomiscosin B was achieved on an RP(18) column using a solvent system consisting of a mixture of acetonitrile-methanol (1:2, v/v) and acetonitrile-water-formic acid (5:95:0.3, v/v) as a mobile phase in a gradient elution mode. A multiple-reaction monitoring (MRM) method was developed for quantification of cleomiscosin A and cleomiscosin B in the seed extracts of Cleome viscosa. On the basis of signal-to-noise ratio of 3, the limit of detection in MRM mode for cleomiscosin A and cleomiscosin B were 1.0 and 4.0 ng/mL respectively. The method was validated in terms of linearity, accuracy and precision for 6 days. The method developed was found to be useful for identification and quantification of cleomiscosin A and cleomiscosin B in the different extracts of the seeds of Cleome viscosa.     

High-performance liquid chromatographic method for identification and quantification of two isomeric coumarinolignoids-cleomiscosin A and cleomiscosin B-in extracts of Cleome viscosa

A simple, accurate and reproducible reverse-phase HPLC method has been developed for identification and quantification of two isomeric coumarinolignoids, cleomiscosin A and B in different extracts of the seeds of Cleome viscosa using photodiode array detection at 326 nm. Cleomiscosin A and B were separated on a Waters symmetry C(18) column (250 x 4.6 mm with 5.0 microm particle size) with an isocratic elution system composed of acetonitrile-methanol (1:2, v/v) and acetic acid-water (0.5:99.5, v/v) in the ratio of 40:60 (v/v). The calibration curves were linear (r(2) > 0.997) in the concentration ranges of 20-100 microg/mL for both compounds. The limits of detection and quantification were 15 and 20 microg/mL for both cleomiscosin A and B. The intra- and inter-day precisions were 3.68 and 2.22% for cleomiscosin A and 4.22 and 5.06% for cleomiscosin B. The recoveries measured at two different concentration levels varied from 98.03 to 110.06%. The method was used to identify and quantify cleomiscosins A and B in different extracts of Cleome viscosa seeds.     

High‐performance liquid chromatographic method for identification and quantification of three potent liver protective coumarinolignoids—cleomiscosin A,cleomiscosin B and cleomiscosin C—in extracts of Cleome viscosa

A simple, rapid, accurate and reproducible reverse‐phase HPLC method has been developed for simultaneous identification and quantification of three coumarinolignoids, cleomiscosin A (Cliv A), cleomiscosin B (Cliv B) and cleomiscosin C (Cliv C) in different extracts of the seeds of Cleome viscosa using photodiode array detection at 326 nm. Cliv A, B and C were separated on a Waters symmetry C₁₈ column (250 × 4.6 mm, 5 μm) using the solvent system consisting of a mixture of acetonitrile : methanol (1:2 v/v) and water : acetic acid (99.5:0.5 v/v) as a mobile phase in a gradient elution mode. The calibration curves were linear in the concentration ranges 15–200, 10–80 and 15–180 μg/mL for Cliv A, Cliv B and Cliv C, respectively. The limits of detection and quantification for Cliv A, Cliv B and Cliv C were 15 and 20 μg/mL, 10 and 15 μg/mL and 15 and 20 μg/mL, respectively. The intra‐day and inter‐day precisions were 2.08 and 0.93% for Cliv A , 1.22 and 0.39% for Cliv B and 1.29 and 0.23 for Cliv C respectively. The developed HPLC method was used to identify and quantify Cliv A, Cliv B and Cliv C in different extracts of seed of Cleome viscosa. Copyright © 2010 John Wiley & Sons, Ltd.     

A New Trinortriterpenoid from Cleome chrysantha

PreviousstudiesonchemicalconstitutionsoftheCleomespecieshaveuncoresedthepresenceoftriterpenoids,nor-triterpenoids,coumarino-lignansandflavanoidsl4.ButtherehasnotbeenanyreportsonthespeciesCleomechrysantha.InourinvestigationontheEgyptianCleomespecies,weisolatedanewtrinortriterpenoid,l-epibrachycarpone1fromtheaerialpartsofCleomechrysantha,togetherwith0-sitosterol2anddaucosterol3.Thispaperdealswiththestructuralelucidationof1.1wasobtainedascolorlesscrystals,mp274--276"C.ItsElMSgavea[M] peakatI…