Isolation of Terpenoids from Trichilia quadrijuga (Meliaceae) by Droplet Counter-Current Chromatography

The Trichilia genus (Meliaceae) consists of about 230 species distributed throughout tropical America and its phytochemical profile is rich in terpenic metabolites. Droplet counter-current chromatography (DCCC) was used for the isolation and purification of secondary metabolites obtained from a dichloromethane fraction (2.9 g) of Trichilia quadrijuga stems. A hexane–ethyl acetate–methanol–water (1:2:1.75:1, v/v/v/v) solvent system was chosen to isolate 2β,3β,4β-trihydroxypregnan-16-one (1, 24.5 mg, 0.8%), kudtdiol (2, 45.0 mg, 1.6%) and 3-O-β-d-glucopyranosylsitosterol (3, 6.0 mg, 0.2%). The results showed that DCCC was a very effective tool for the isolation of terpenes from T. quadrijuga.     

Validation assay for total flavonoids, as rutin equivalents, from Trichilia catigua Adr. Juss (Meliaceae) and Ptychopetalum olacoides Bentham (Olacaceae) commercial extract

An ultraviolet spectrophotometric method was validated for total flavonoid quantitation, as rutin equivalents, present in the Trichilia catigua Adr. Juss (Meliaceae) and Ptychopetalum olacoides Bentham (Olacaceae) commercial extract. Parameters as linearity, interval (range), specificity, estimated limit of detection (LOD, microg/mL), estimated limit of quantitation (LOQ, microg/mL), recovery (R, %), precision or relative standard deviation (RSD, %), and accuracy (E, %) were established. The analytical method was validated according to the experimental results: correlation coefficient (r = 0.9997); interval (RSD = 0.15-0.47%; E = 98.98-101.24%); specificity to total flavonoids quantitation, as rutin equivalents, at wavelength 361.0 nm; LOD = 0.09 microg/mL and LOQ = 0.27 microg/mL; R = 99.36-102.14%; adequate intra- and interrun precision (0.30-0.49% and 0.31-0.81%), and intra- and interrun accuracy (100.60-102.38% and 98.58-100.38%).     

Rubescins I and J,further limonoid derivatives from the stem bark of Trichilia rubescens (Meliaceae)

Two new tetranorterpenoid derivatives named rubescins I (1) and J (2), were isolated along with six known compounds including rubescin D (3), lichexanthone (4), scopoletin (5), scopoletin O-glycoside (6), β-sitosterol (7) and stigmasterol (8) from the stem bark of Trichilia rubescens (Meliaceae). The structures of the compounds were determined by means of MS, different NMR and by comparison with related data reported in the literature.     

A validated higher-performance liquid chromatography method for quantification of cinchonain Ib in bark and phytopharmaceuticals of Trichilia catigua used as Catuaba

The hydroalcoholic extract, prepared from authentic chopped barks of Trichilia catigua, was evaluated by high-performance liquid chromatography using a diode array detector (200-400 mn). The crude extract was purified by rotation locular counter-current chromatography and the chloroform fraction obtained was clean-up by solid-phase extraction. With the aim of getting preliminary structure information on-line, the methanol fraction thus obtained was analyzed by gradient elution using the diode array detector coupled to a mass spectrometer. The presence of flavalignan in this extract was inferred by the chromatographic band, in the total ion current trace, that had an [M-H](-) = 451. With this information, cinchonain Ib was isolated as a pure compound from the crude hydroalcoholic extract using a solid-phase extraction procedure for the sample clean-up followed by a semi-preparative separation using the reverse mode of elution. The isolated compound, after complete characterization, was used as an external standard for the development and validation of a method for the analysis of this compound in herbal medicines using the ultraviolet as the detector. The validated method has been successfully applied for quantification of cinchonain Ib in commercialized herbal medicines sold as Catuaba in Brazil and also in standard chopped barks of T. catigua.     

Synthesis of fused tricyclic gamma-lactones mediated by manganese(III) acetate

Exposure of cyclic alkenes bearing a carboxylic acid and a malonate group to manganese(III) acetate and an appropriate copper(II) salt provides the corresponding tricyclic gamma-lactones in good yield.     

Corrected Structure of Aglalactone Isolated from Aglaia elaeagnoidea (Meliaceae)

Summary.  The structure of aglalactone, a dihydrobenzofuranone from Aglaia elaeagnoidea, was revised on the basis of 2D NMR data and lanthanide induced shifts (LIS). Received June 29, 2000. Accepted July 11, 2000     

Alkaloids from the stem bark of Turraeanthus africanus (Meliaceae)

Fractionation of the methanol extract of the stem bark of Turraeanthus africanus led to the isolation of two new alkaloids designated turraeanthin A and B, together with two known alkaloids. The structures of the new alkaloids were elucidated by means of spectroscopic analysis and characterized as 10-O-demethyl-17-O-methyl isoarnottianamide and 11-demethoxyl-12-methoxyl oxynitidine respectively.     

Corrected Structure of Aglalactone Isolated from Aglaia elaeagnoidea (Meliaceae)

 The structure of aglalactone, a dihydrobenzofuranone from Aglaia elaeagnoidea, was revised on the basis of 2D NMR data and lanthanide induced shifts (LIS).     

Separation of Phenols from the Leaves of Toona Sinensis (Meliaceae) by Capillary Electrophoresis

A micellar electrokinetic capillary chromatography (MEKC) method, using UV detection, was developed for the determination of polyphenols in Toona sinensis (Meliaceae); the procedure involved precipitation of polyphenols from the leaves of T. sinensis using methanol. The structures can be established with fifteen compounds including methyl gallate, gallic acid, kaempferol, quercitin, quercitrin, rutin, kaempferol‐glucoside, catechin, epicatechin, stearic acid, palmitic acid, β‐sitosterol, stigmasterol, β‐sitosteryl‐glucoside, and stigmasteryl‐glucoside by spectroscopic analysis. However, there has been no investigation to quantitate the polyphenols that form T. sinensis. Thus, seven polyphenols of T. sinensis with UV absorbance, catechin (C), epicatechin (EC), methyl gallate (MG), rutin (R), gallic acid (G), quercitrin (Q), and kaempferol (K) were separated within 10 min with a 40 cm uncoated fused‐silica column, with the RSD < 3% (migration times), voltage at 15 kV using this method. On‐column detection was carried out at 254 nm. The detection limit of this method for all analytes ranged from 19.5 to 0.02 μM (RSD < 3.1%). The method provided a rapid and sensitive identification of polyphenols of interest in T. sinensis and is suitable for biological activity studies.     

Structure determination by nmr of azadirachtin and related compounds from azadirachta indica a. Juss (Meliaceae)

Two novel limonoids 3-deacetyl-3-cinnamoylazadirachtin (6) and 1-tigloyl-3-acetyl-11-methoxyazadirachtinin (8) have been isolated from Azadirachta indica extracts in addition to azadirachtin (4), 22,23-dihydro-23β-methoxyazadirachtin (5) and 3-tigloylazadirachtol (7). The assignment of the structures (5) - (8) and the reassignment of the azadirachtin structure as (4) was achieved on the basis of detailed ¹H and ¹³C n.m.r. spectroscopic analyses.     

UV spectrophotometric determination of bioflavonoids from a semisolid pharmaceutical dosage form containing Trichilia catigua Adr. Juss and Ptychopetalum olacoides Bentham standardized extract: analytical method validation and statistical procedures

A precise, accurate, and sensitive UV spectrophotometric method was developed and validated for routine quantification of total bioflavonoids, expressed as rutin, from a topical oil-in-water pharmaceutical emulsion containing the extract of Trichilia catigua Adr. Juss and Ptychopetalum olacoides Bentham. The method was validated experimentally, and the data were treated rigorously by statistical analysis. The following analytical parameters were assessed: linearity, specificity, intra- and interrun precision measured as relative standard deviation (RSD, %), intra- and interrun accuracy (E, %), recovery (Rec., %), limit of detection (LOD, microg/mL), and limit of quantification (LOQ, microg/mL). The UV spectrophotometric method was linear (r = 0.9995) for standard rutin over the concentration range of 5.0-15.0 microg/mL with specificity for total bioflavonoids (expressed as rutin) at 361.0 nm with an absence of interferents from the complex matrix; RSD of < or = 1.79%, intrarun (E = 97.88 +/- 1.75 to 99.0 +/- 0.33%) and interrun (E = 98.38 +/- 1.12 to 100.79 +/- 1.30%) accuracy; Rec. = 98.64 +/- 0.42 to 100.74 +/- 0.41%; LOD = 0.20 microg/mL; and LOQ = 0.30 microg/mL.     

New Minor Cyclopeptides from Arenaria oreophila （Caryophyllaceae）

New minor cyclic peptides, named arenariphilin C 1 and arenariphilin D 2, were isolated from the whole plants of Arenaria oreophila (Caryophyllaceae). Their structures were determined as cyclo(Pro1-Pro2-Leu1-Leu2-Phe-Ser-Gly-Thr) and cyclo (Ser-Cys) on the basis of spectroscopic data, especially by two-dimension NMR (2D NMR) technologies.     

Antifeedant triterpenoids from the seeds and bark of Lansium domesticum cv Kokossan (Meliaceae)

Two tetranortriterpenoids, kokosanolide A (1) and C (2) were isolated from the seeds and three onoceranoid-type triterpenoids: kokosanolide B (3), 8,14-secogammacera-7,14-diene-3,21-dione (4) and a 1.5:0.5 mixture of 8,14-secogammacera-7,14(27)-diene-3,21-dione (5) and compound 4 were isolated from the bark of kokossan (Lansium domesticum). Complete 1H- and 13C-NMR data of the triterpenoids 1-5 are reported. The triterpenoids' structures were elucidated primarily by means of high field 1D- and 2D-NMR, IR and HRMS spectral data. Triterpenoids 1-5 exhibited moderate to strong antifeedant activity against the fourth instar larvae of Epilachna vigintioctopunctata.     

Cytotoxicity and Synergistic Effect of the Constituents from Roots of Aglaia odorata (Meliaceae)

Twelve compounds were isolated from the roots of Aglaia odorata. Their structures were established on the basis of NMR and MS data as rocaglaol (1), rocaglamide (2), eichlerialactone (3), sapelins A (4), isofouquierone (5), eichlerianic acid (6), shoreic acid (7), agladupol E (8), 3-epimeliantriol (9), cleomiscosins B (10), 2β,3β-dihydroxy-5α-pregnane-16-one (11) and β-d-glucopyranos-1-yl N-methylpyrrole-2-carboxylate (12). Among them, compounds 1 and 2 showed significant cytotoxicity against human cancer cell (HL-60, SMMC-7721, A-549, MCF-7 and SW480) with IC₅₀ values of 0.007–0.095 μM, while compounds 3–5 and 10 and 11 showed moderate to no cytotoxicity (IC₅₀ 0.43 to values >40 μM). Compound 6 showed only weak cytotoxicity (IC₅₀ 6.87 to >40 μM) and its epmier 7 was completely inactivite (IC₅₀>40 μM) in the assay. However, potent synergistic effect was observed when the molar ratio of 6 to 7 is between 4:1 and 1:1.     

Open tubular supercritical fluid chromatography of triterpene acids from Dysoxylum pettigrewianum (Meliaceae)

Open tubular supercritical fluid chromatography (SFC) with carben dioxide as the mobile phase has been investigated for the separation of four triterpene acids, two of which were structural isomers. These compounds, isolated from Dysoxylum pettigrewianum (Meliaceae), were chromatographed on three stationary phases with different selectivities. 30%-biphenylmethylpolysiloxane and polyethylene glycol (Carbowax 20M) phases exhibited poor chromatographic selectivity for the compounds. The separation was improved by employing the shape selectivity of a liquid crystalline biphenylcarboxylate ester polysiloxane and utilizing simultaneous temperature and pressure programming. The elution order was explained on the basis of the molecular structures of the triterpene acids, and their interaction with the liquid crystal phase.     

Dammarane-Type Triterpenoid from the Stem Bark of Aglaia elliptica (Meliaceae) and Its Cytotoxic Activities

Two new dammarane-type triterpenoid fatty acid ester derivatives, 3β-oleate-20S-hydroxydammar-24-en (1) and 3β-oleate-20S,24S-epoxy-25-hydroxydammarane (2) with a known dammarane-type triterpenoid compound, such as 20S-hydroxydammar-24-en-3-on (3), were isolated from the stem bark of Aglaia elliptica (C.DC.) Blume. The chemical structures were determined by spectroscopic methods, including FTIR, NMR (one and two-dimensional), and HRESITOF-MS analysis, as well as chemical derivatization and comparison with previous literature. Furthermore, the synthetic analog resulting from transesterification of 1 and 2 also obtained 3β,20S-dihydroxy-dammar-24-en (4) and 20S,24S-epoxy-3β,25-dihydroxydammarane (5), respectively. The cytotoxic effect of all isolated and synthetic analog compounds was evaluated using PrestoBlue reagent against MCF-7 breast cancer cell and B16-F10 melanoma cell lines. The 20S-hydroxydammar-24-en-3-on (3) showed the strongest activity against MCF-7 breast cancer and B16-F10 melanoma cell, indicating that the ketone group at C-3 in 3 plays an essential role in the cytotoxicity of dammarane-type triterpenoid. On the other hand, compounds 1 and 2 had very weak cytotoxic activity against the two cell lines, indicating the presence of fatty acid, significantly decreasing cytotoxic activity. This showed the significance of the discovery to investigate the essential structural feature in dammarane-type triterpenoid, specifically for the future development of anticancer drugs.     

Vinyl and carbene ruthenium(II) complexes from hydridoruthenium(II) precursors

The reactions of the hydrido compounds [RuHCl(CO)(L)2][L = PiPr3 (1), PCy3 (2)] with HC(triple bond)CR (R = H, Ph, tBu) afforded by insertion of the alkyne into the Ru-H bond the corresponding vinyl complexes [RuCl(CHCHR)(CO)(L)2], 3-8, which upon protonation with HBF4 gave the cationic five-coordinated ruthenium carbenes [RuCl(CHCH2R)(CO)(L)2]BF4, 9-14. Subsequent reactions of the carbene complexes with PR3(R = Me, iPr) and CH3CN led either to deprotonation and re-generation of the vinyl compounds or to cleavage of the ruthenium-carbene bond and the formation of the six-coordinated complexes [RuCl(CO)(CH3CN)2(PiPr3)2]BF4, 17, and [RuH(CO)(CH3CN)2(PiPr3)2]X, 18a,b. The acetato derivative [RuH(2-O2CCH3)(CO)(PCy3)2], 19, also reacted with acetylene and phenylacetylene by insertion to yield the related vinyl complexes [Ru(CHCHR)(kappa2-O2CCH3)(CO)(PCy3)2], 20, 21, of which that with R = H was protonated with HBF4 to yield the corresponding cationic ruthenium carbene 22. With [RuHCl(H2)(PCy3)2], 25, as the starting material, the five-coordinated chloro(hydrido)ruthenium(II) compounds [RuHCl(PCy3)(dppf)], 26(dppf = [Fe(eta5-C5H4PPh2)2]), [RuHCl[Sb(CH2Ph)3](PCy3)2], 27, and [RuHCl(CH3CN)(PCy3)2], 30, were prepared. The reactions of 27 with HCCR (R = H, Ph) gave the hydrido(vinylidene) complexes [RuHCl(CCHR)(PCy3)2], 28 and 29, whereas treatment of 30 with HC(triple bond)CPh afforded the vinyl compound [RuCl(CHCHPh)(CH3CN)(PCy3)2], 31. The molecular structures of 11(R = tBu, L = PiPr3) and 26 were determined crystallographically.     

Synthesis of chiral 3-substituted gamma-lactones and 9-furanosyladenine from (R)-2-(2,2-diethoxyethyl)-1,3-propanediol monoacetate prepared by lipase-catalyzed reaction

A chiral building block, (R)-2-(2,2-diethoxyethyl)-1,3-propanediol monoacetate was synthesized in high optical and chemical yields by lipase-catalyzed transesterification. From this compound, we synthesized chiral 3-substituted gamma-lactones and a new nucleoside with antiviral activity.