Chloroform-methanol Mixture Research Articles

Phytochemical and Anti-Microbial Analysis of the Roots of Ficus Exasparata (Anwirinwa)

The roots of Ficus exasparata of the Moraceae family were duly analysed to ascertain the active constituents responsible for the use of the plant part in the treatment of inflammatory diseases, treatment of internal bleeding, bruises, cough and dangerous boils among others. The phytochemical analysis showed the presence of some secondary metabolites in various concentrations with saponin and alkaloids present in low concentration while reducing sugars were present in very high concentration with protein, carbohydrates and acidic components totally absent. The Atomic Absorption Spectroscopic (AAS) analysis showed the presence of certain trace elements such as Cd (0.55ppm), Cr (1.88ppm),As (0.38ppm), Hg (0.09ppm) etc which were higher than the WHO recommended standard of Cd (0.003ppm), Cr (0.05ppm), As (0.01ppm), Hg (0.001ppm) respectively an observation that could be attributed to the crude nature of the plant parts. Three different choice solvents (Chloroform, Ethylacetate and Chloroform-Methanol mixture) were used in the extraction of the plant part. The extracts were subsequently subjected to thin layer chromatography with respective Rf values as 0.13, 0.22 and 0.60 respectively showing just a single spot each for all the solvents used. The extracts were subjected to antimicrobial screening using eight pathogenic bacteria species and three fungi with only the chloroform extract exhibiting activity on the selected test organisms. The three extracts from the sample were subjected to structural elucidation using a combination of some spectroscopic techniques such as FTIR, UV-VIS, H1-NMR, C13-NMR and GCMS. The spectral analysis suggested the presence of Coumarin-3-carboxamide, 8-allyl-N-(3-nitrophenyl) and 1,2,3-propanetricarboxylic acid, 2-(acetyloxy)-tributyl ester for chloroform and ethyl acetate extracts respectively. The constituents of these extracts such as Coumarin in chloroform extract could account for its potency in curing certain illnesses as coumarin is a well known natural product that has displayed a broad range of biological activities.

Cholesterol and Fatty Acids Oxidation in Meat from Three Muscles of Massese Suckling Lambs Slaughtered at Different Weights

Eighteen Massese male lambs fed mainly with maternal milk were slaughtered at 11, 14 and 17 kg. Samples of Longissimus dorsi (LD), Triceps brachii (TB) and Semimembranosus (Sm) muscles were collected. Total intramuscular lipids were extracted by means of a mixture of chloroform methanol 2/1. Cholesterol content and its oxidation product (COP) were determined by a gas chromatography apparatus equipped with an apolar 30 m column. Fatty acid oxidation was evaluated by means of thio-barbituric acid reactive substances (TBARS) extracting the sample with aqueous acidic solution. The effect of slaughter weight on oxidation of intramuscular lipids was found only in TB muscles. In this muscle the cholesterol content showed a decreasing trend, while the content of COPs significantly increased with the age of animals. Among the COPs, the 7-ketocholesterol and 7β-hydroxycholesterol were the most abundant, followed by α- and β-epoxy-cholesterol and cholestan-triol. The content of TBARS did not vary owing to a similar fatty acid composition of intramuscular fat across weight of slaughter. In any case, the values of TBARS did not reach the threshold of the detection of off-flavour in meat.

Sample pretreatment of brain tissues and cerebrospinal fluid for NMR investigations

Samples of rat homogenated brain cortex and cerebrospinal fluid prepared by various protocols were investigated by high-resolution NMR. The stability of the major metabolites, biomarkers of neurode-generative diseases, was examined depending on the procedure of sample pretreatment. As shown, the optimal conditions of the sample pretreatment of brain tissues to NMR analysis involve mechanical homogenization followed by the addition of deuterated water and for prolong experiments, an extraction with a methanol-chloroform mixture (2: 1). Samples of cerebrospinal fluid for NMR study should be prepared in buffer media based on deuterated water to improve the stability of the preparation and prevent the drift of metabolite signals in the spectra of different samples during the experimentation. The chosen optimal conditions of sample pretreatment of brain tissue and cerebrospinal fluid can be used in future to study changes in the composition and concentration of metabolites in neurodegenerative diseases by virtue of NMR. The proposed methods allow more accurate the diagnosis of pathologies of the brain function at early stages.

Solvent-dependent Assembly of Discrete and Continuous CoCl2 Adamantane-based Ligand Complexes: Observations by CSI–Mass Spectrometry and X-ray Crystallography

Discrete and continuous coordination structures were obtained in single crystals of CoCl₂ and an adamantane-based bidentate ligand bearing imidazolyl groups, depending on the methanol concentration in a methanol-chloroform mixture. Single-crystal X-ray structure analysis revealed that the metal centers exhibited a tetrahedral geometry in the discrete complex and an octahedral geometry in the continuous metal complex. Conventional analytical methods, including UV-vis and NMR spectroscopy, could not identify those two complexes in solution. In contrast, cold-spray ionization mass spectrometry could detect differences between the discrete complex and the continuous metal complex, and ion peaks due to continuous ligand adducts were found only in the spectrum of the continuous metal complex.

Selective Synthesis of Glyceryl Tris[9,10‐(threo)‐dihydroxyoctadecanoate

AbstractElaborating novel triacylglyceride (TAG) based polyester hyperbranched unimolecular encapsulating agents represents an original and promising approach to the selective delivery of hydrophobic biologically active compounds. However, selective modification of double bonds in unsaturated TAG to obtain corresponding pure polyhydroxy derivatives with high yields is still a big challenge. Two novel approaches to synthesize the glyceryl tris[9,10‐(threo)‐dihydroxyoctadecanoate] were proposed and tested: (1) via the bromination and nucleophilic substitution of secondary halide functions by oxyacetyl groups followed by the hydrolysis of acetyls, and (2) direct transformation of the double bonds by the reaction with peroxoformic acid in an excess of formic acid, and the removal of formyl protective groups. Glycerol trioleate and natural olive oil were used as starting materials. The first synthetic route allowed for successful preparation of the corresponding polyhalide and polyoxyacetyl products; however, a more effective final deacetylation procedure is required. The second proposed approach showed a very good reproducibility in obtaining hydroxy‐oxyformyl derivatives. The optimal conditions of the reaction involve the use of diethyl ether as a cosolvent and stirring at room temperature for 30 min. To remove the formyl groups, three original procedures using organic solvent medium at room temperature were proposed: in the presence of cesium carbonate in chloroform–methanol mixture, and two methods using hydrochloric acid in chloroform–methanol mixture or in acetone. All three methods were efficient to carry out the deformylation; nevertheless, TAG esters remained stable only with 15–17 % hydrochloric acid in acetone. Simple isolation procedures and high overall yields (95.6 and 94.9 % for both triolein and olive oil, respectively) allow considering the second approach as a promising method to obtain threo‐polyhydroxy derivatives from unsaturated TAG.

Studies on the seasonal variations in the proximate composition of ascidians from the Palk Bay, Southeast coast of India

ObjectiveTo investigate the seasonal fluctuations of the proximate composition of the ascidians muscle. MethodsThe moisture content was estimated by drying 1 g of fresh tissue at a constant temperature at 105°C for 24 h. The loss of weight was taken as moisture content. The total protein was estimated using the Biuret method. The total carbohydrate in dried sample was estimated spectrophotometrically following the phenol- sulphuric acid method. The lipid in the dried sample tissue was gravimetrically estimated following the chloroform-methanol mixture method. Ash content was determined gravimetrically by incinerating 1 g dried sample in muffle furnace at about 550°C for 6 h and results are expressed in percentage. ResultsIt was found very difficult to compare the monthly variations, as all the ten species, exhibited wide fluctuations in their proximate compositions. For the sake of convenience, average seasonal values were calculated by summing the monthly values. ConclusionThe proximate composition of the 10 commonly available ascidians showed high nutritive value and hence these groups especially solitary ascidians can be recommended for human consumption in terms of pickles, soup, curry and others after ensuring the safety of consumers.

A handbook for uncovering the complete energetic budget in insects: the van Handel's method (1985) revisited

Insects comprise relevant biological models for investigating nutrient acquisition and allocation processes in the context of life‐history ecology and evolution. However, empirical investigations are still partly limited by the lack of availability of simple methods for simultaneously estimating the four major energetic components (i.e. lipids, free sugars, glycogen and proteins) in the same individual. In the present work, we validate a fast, reproducible and cheap method for overcoming this problem that uses different solvents successively. First, proteins are solubilized in a phosphate‐lysis buffer and then quantified according to the classical Bradford assay procedure. In a second step, a chloroform–methanol mixture is added to the aqueous phase, which allows assay of the total lipid fraction, as well as the free sugars and glycogen in the same insect homogenate. In addition, a micro‐separation procedure is adapted to partition the total lipids into neutral (mainly stored lipids) and polar (mainly structural lipids) components. Although these assays are conducted sequentially in the same individual, the sensitivity of our method remains high: the estimated amount of each energetic compartment does not differ from that obtained with former, partial methods. Our method should thus largely improve our knowledge about nutrient acquisition and allocation among insects not only in laboratory‐reared individuals, but also in animals caught in the wild. Descriptions and recommendations are given at each step of the protocol to adapt the procedure to various insect species. Finally, to prevent misinterpretation of data generated in accordance with this protocol, the limits of our method are discussed in the light of life‐history studies.

Freeze-dried ABTS+ method: A ready-to-use radical powder to assess antioxidant capacity of vegetable oils

Abstract A new approach to 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonate) (ABTS +) assay was conducted. ABTS + radical solution in water was obtained by using classical method of incubating ABTS with potassium peroxodisulphate. The resulting radical solution was freeze-dried and tested for its radical efficiency by comparing freshly prepared ABTS + (FP-ABTS) via using standard antioxidants. A high correlation was observed between the results obtained from FP-ABTS and freeze-dried ABTS (FD-ABTS) assays. When it was stored in the freezer, FD-ABTS powder was found to keep its initial activity to a great extend over one month. For the measurement of antiradical capacity of oil samples, FD-ABTS was dissolved in methanol and diluted to desired concentrations with a chloroform–methanol mixture. Linearity and accuracy of the method was confirmed by measuring the antiradical capacity of stripped sunflower oil spiked with different concentrations of standard antioxidants. Antiradical capacity of 22 oil samples including some commonly consumed vegetable oils was also tested by the FD-ABTS method.

Quality of Canola Oil Obtained by Conventional and Supercritical Fluid Extraction

Quality of canola oil obtained by the supercritical fluid extraction (SFE), using CO2 with ethanol as a co-solvent, was evaluated and compared to that of the conventionally-obtained oils using either n-hexane or chloroform methanol mixture. Physical characteristics, chemical properties, fatty acid composition and phenolic profile of oils were investigated. The SFE oil showed significantly lower melting point, peroxide value (PV) and higher free fatty acids (FFAs) and iodine value (IV) than the n-hexane-extracted one. There were no significant differences in the fatty acid composition of different oils. The SFE oil showed significantly higher phenolic content (35.91, 10.15, 3.16, 0.32 and47.48mg/g of sinapic acid, sinapine, sinapoyl glucose, canolol and total phenolics) as compared to 0.08, 0.70, 0.88, 0.45 and0.71mg/g, respectively in the n-hexane-extracted oil. These results indicate the superiority of SFE and advocate its use for the extraction of highly stable and functional canola oil for further health and nutraceutical uses. The present results have an industrial and technological relevance as SFE could be competitive with the traditional extraction techniques providing an environmental approach and enhancing the obtained oil quality and stability. After recovery of the initial installation costs, SFE could be more economic than conventional extraction. However, further economical studies are needed to validate this last conclusion.

Enzymological analysis of the tumor suppressor A-C1 reveals a novel group of phospholipid-metabolizing enzymes

A-C1 protein is the product of a tumor suppressor gene negatively regulating the oncogene Ras and belongs to the HRASLS (HRAS-like suppressor) subfamily. We recently found that four members of this subfamily expressed in human tissues function as phospholipid-metabolizing enzymes. Here we examined a possible enzyme activity of A-C1. The homogenates of COS-7 cells overexpressing recombinant A-C1s from human, mouse, and rat showed a phospholipase A½ (PLA½) activity toward phosphatidylcholine (PC). This finding was confirmed with the purified A-C1. The activity was Ca²⁺ independent, and dithiothreitol and Nonidet P-40 were indispensable for full activity. Phosphatidylethanolamine (PE) was also a substrate and the phospholipase A₁ (PLA₁) activity was dominant over the PLA₂ activity. Furthermore, the protein exhibited acyltransferase activities transferring an acyl group of PCs to the amino group of PEs and the hydroxyl group of lyso PCs. As for tissue distribution in human, mouse, and rat, A-C1 mRNA was abundantly expressed in testis, skeletal muscle, brain, and heart. These results demonstrate that A-C1 is a novel phospholipid-metabolizing enzyme. Moreover, the fact that all five members of the HRASLS subfamily, including A-C1, show similar catalytic properties strongly suggests that these proteins constitute a new class of enzymes showing PLA½ and acyltransferase activities.

Isolation and Characterization of Bioenhancing Catechins from Thea sinensis Leaves

Thea sinensis syn. Camellia sinensis (Theaceae) is an evergreen shrub indigenous to India. The leaves of this tea plant have been used both as a social and medicinal beverage since 3000 BC [1]. Traditionally green tea is recommended for headaches, body aches and pains, digestion, enhancement of immune defenses, detoxification, as an energizer and to prolong life [2]. The leaves have more than 700 chemical constituents, among which flavanoids, amino acids, vitamins, caffeine and polysaccharides are important to human health. In general, fresh tea leaves contain polyphenols, among which catechins prevail [3]. In the present study, phytochemical and biological screening of leaves of T. sinensis was undertaken. Column chromatography of methanolic extract with chloroform-methanol mixture in the increasing order of polarity yielded six catechins characterized as eremphilanyl gallate, epigallocatechin galactosidic tridecadiene, dimethoxycatechin gallate diglucosidic decane, protocatechinic acid triglycosidic undecane, protocatechinic acid tetraglycosidic decane and protocatechinic acid tetraglycosidic octane on the basis of NMR, ESI-MS, FTIR and UV data. The methanolic extract of tea leaves, at concentrations (3% and 6% w/v), were evaluated as permeation enhancers for amoxicillin using everted rat sac model. amoxicillin is a class III drug with low permeability. The amount of Amoxicillin that traversed the gut was followed spectrophotometrically at 254nm. The extract showed significant increase in its availability at both the concentrations (P<0.05). It has recently been reported that some catechins of green tea enhanced the effect of β-lactams [4]. Here it is postulated that catechins may be responsible for enhanced activity of β-lactams by increasing its availability.

Synthesis and spectral properties of helicate of cobalt(II) with Bis(1,2,3,7,9-pentamethyldipyrrolylmethen-3-yl)methane

Reaction of cobalt(II) acetate tetrahydrate with compound I in a chloroform–methanol mixture in the presence of triethylamine afforded a binuclear homoleptic double-helical compound [Co2L2] II (Scheme 2). The electron absorption spectra of [Co2L2] solutions in organic solvents (С6Н6, DMF, CHCl3) contain two bands. Depending on the nature of the solvent, the most intensive band has a maximum in the range of 524–528 nm with a shoulder on the left slope at 490– 493 nm; a broadened less intense band of the charge transfer is in the range of 373–374 nm. Type of the electron absorption spectrum of helicate [Co2L2] in the organic solvents is consistent with the published data for the corresponding by composition Co(II) helicate with the structurally related bis(dipyrrolylmethenes) [4, 5]. In a non-polar inert benzene instead of the polar solvents, the electron-donating DMF and protonDOI: 10.1134/S1070363211010324

Synthesis and complexive abilities in cone conformation of upper rim functionalized of calix[4]arene with alkali and alkali earth cations

The synthesis and complexive abilities of 5-mono­[(tert-butoxycarbonyl)-amino]-11,17,23-triamino-25,26,27,28-tetrabutoxycalix[4]arene (L1) and 5,11,17,23-tetrakis[(tert-butoxycarbonyl)-amino]-25,26,27,28-tetrabutoxycalix[4]arene (L2) towards the alkali cations Li+, Na+, K+, Rb+, Cs+ and the alkali earth cations Mg+2, Ca+2, Sr+2 and Ba+2in methanol-chloroform mixture have been evaluated at 25°C using UV-Vis spectrophotometric techniques. The results show that the ligands are capable of complexing with all the cations by 1:1 metal to ligand ratios. Molecular mechanics calculations have been used to model the geometries of the alkali and alkali earth cations with the ligands using the Hyperchem 7 computational package. The selectivity is presented considering that the calculated formation constants are in the order Li+>>Na+>K+>Rb+>Cs+and Mg+2>Ca+2>Sr+2>Ba+2with the ligands. Also, L1 shows more attraction towards the smaller alkali and alkali earth metal ions than L2.

Lipid Composition of Walnut (Juglans regia L.) Cv. Pedro Grown in Bulgaria

SUMMARY Lipid composition of walnut ( Juglans regia L.) cv. Pedro grown in Bulgaria (harvest year 2009) has been investigated concerning the neu tral lipid classes, their fatty acid composition and the triacylglycerol species. Fat co ntent of the oil (71.3%) was determined gravimetrically after extraction of dry kernels in Soxhlet apparatus (6 h with hexane). The walnut oil for analyses of lipids was obtained from kernels by a procedure included firstly treatment of walnuts with iso-propanol and then ext raction by chloroform-methanol (2:1) mixture (Christie, 1993). Lipid classes were separa ted by thin layer chromatography (silica gel G plates, developed with hexane-acetone, 10:1 v /v), identified towards a standard mixture, isolated and weighed. The walnut cultivar Pedro was found to contain the following lipid classes: polar lipids (0.5%), diacylglycerols (1.7% ), sterols (0.5%), unidentified lipid zone X (0.7%), triacylglycerols (95.8%) and sterol esters (0.8%). Each of these classes was transmethylated according to Christie (2003) and th e methyl esters were analysed by gas chromatography. The results are given in Tab 1.

ChemInform Abstract: Recognition by New Symmetrically Substituted Chiral Diphenyl- and Di- tert-butylpyridino-18-crown-6 and Asymmetrically Substituted Chiral Dimethylpyridino-18-crown-6 Ligands of the Enantiomers of Various Organic Ammonium Perchlorates.

Three new chiral pyridino-18-crown-6 ligands have been prepared. These ligands contain either two phenyl, two tert-butyl, or two methyl substituents on chiral macroring carbon atoms. The chiral di-tert-butyl-substituted diester crown analogue was also prepared. The starting chiral di-tert-butyl-substituted tetraethylene glycol needed to prepare the two di-tert-butyl-substituted crowns was obtained from chiral tert-butyl-1,2-ethanediol, which was resolved from its bis(hydrogen phthalate) brucine salt. A high degree of chiral recognition of CD 2 Cl 2 of the enantiomers of [α-(1-naphthyl)ethyl] ammonium perchlorate (NapEt) was shown by the diphenyl- and di-tert-butyl-substituted crowns by differences in the free energy of activation (ΔG c : ) values determined by temperature-dependent i H NMR spectroscopy. The diphenyl- and di-tert-butyl-substituted crowns also exhibited high chiral recognition for the enantiomers of NapEt and other chiral organic ammonium salts in methanol and methanol-chloroform mixtures as shown by a large difference in the log K values determined by a direct i H NMR technique

Chemical composition of the leaves of Periploca sepium

The genus Periploca (Asclepiadaceae) contains 12 species, of which there are four species in China. Periploca sepium Bage, widely distributed in the northeast, southeast, northwest, and southwest of China, is one of the four species. The root of Periploca sepium Bage has been used as a traditional Chinese herbal medicine for more than two thousand years [1]. Previous phytochemical studies of this species have isolated, from the root of P. sepium, cardenoides, pregnane glycosides, etc. [2, 3]. For many years, research on the plant has mainly concentrated on the underground part. To continue searching for novel agents from P. sepium, the leaves of P. sepium were chosen for phytochemical investigation. In this paper, we report the isolation and identification of four compounds. The leaves of P. sepium Bage were collected from Qinling Mountain, Shaanxi, China, in July, 2006 and identified by Prof. E. X. Han of Northwest A & F University. The voucher specimen was deposited in the Forestry College, Northwest A & F University. Air-dried and powdered leaf of P. sepium Bage (3.9 kg) was extracted exhaustively with 95% ethanol at room temperature. The ethanol extract was concentrated in vacuum at 60 C to give a residue. The residue was suspended in water and successively extracted with petroleum ether, ethyl acetate, and n-butanol. The ethyl acetate extract (47.0 g) was subjected to repeated silica gel column chromatography with Sephadex LH-20 using different percentages of chloroform–methanol mixtures, and compounds 1 and 2 were obtained. Repeated chromatography of the n-butanol extract (47.3 g) on silica gel with chloroform–methanol and Sephadex LH-20 afforded compounds 3 and 4. The structures of these compounds were elucidated by a combination of spectral analysis and were determined as oleanolic acid (1) [4], quercetin (2) [5], 6 -methyl ester of quercetin-3-O-D-glucuronide (3) [6], and isoquercitrin (4) [7] by comparison with the data in the literature. All these four compounds were isolated for the first time from P. sepium, and compound 2 was isolated for the first time from the genus of Periploca. Oleanolic acid (1) white powder, mp > 300 C. IR (KBr, max, cm –1): 3446 (OH), 2941, 2861, 1693 (C=O), 1636 (C=C), 1462, 1385, 1274, 1084, 1036, 749. 1H NMR (300 MHz, CDCl3, , ppm): 0.74 (3H, s, CH3, H-26), 0.77 (3H, s, CH3, H-23), 0.90 (3H, s, CH3, H-24), 0.91 (3H, s, CH3, H-29), 0.92 (3H, s, CH3, H-30), 0.98 (3H, s, CH3, H-25), 1.13 (3H, s, CH3, H-27), 5.28 (1H, br.s, H-12), 3.24 (1H, br.s, H-3), 2.81 (1H, br.s, H-18). Quercetin (2) yellow needle crystals, mp > 300 C. UV (CH3OH, max, nm): 381.0, 257.0, (CH3OH + NaOH, max, nm): 419.0, 330.0, 280.0, (CH3OH + AlCl3, max nm): 448.0, 271.0. 1H NMR (300 MHz, DMSO-d6, , ppm, J/Hz): 6.19 (1H, d, J = 1.8, H-6), 6.41 (1H, d, J = 1.8, H-8), 6.88 (1H, d, J = 8.4, H-5 ), 7.54 (1H, dd, J = 2.0, 8.4, H-6 ), 7.68 (1H, d, J = 2.0, H-2 ), 9.34 (1H, s, 3 -OH), 9.40 (1H, s, 3-OH), 9.62 (1H, s, 4 -OH), 10.80 (1H, s, 7-OH), 12.51 (1H, s, 5-OH). 13C NMR (75 MHz, DMSO-d6, ): 147.24 (C-2), 136.19 (C-3), 176.29 (C-4), 161.17 (C-5), 98.63 (C-6), 164.34 (C-7), 93.79 (C-8), 156.58 (C-9), 103.46 (C-10), 122.40 (C-1 ), 115.51 (C-2 ), 145.51 (C-3 ), 148.15 (C-4 ), 116.05 (C-5 ), 120.42 (C-6 ). 6 -Methyl ester of quercetin-3-O-D-glucuronide (3) yellow needle crystals, mp 181–183 C. 1H NMR (300 MHz, DMSO-d6, , ppm, J/Hz): 6.21 (1H, d, J = 1.1, H-6), 6.41 (1H, d, J = 1.1, H-8), 6.83 (1H, d, J = 8.4, H-5 ), 7.51 (1H, d, J = 2.5, H-2 ), 7.56 (1H, dd, J = 2.5, 8.4, H-6 ), 5.46 (IH, d, J = 6.9, H-1 ), 3.56–3.73 (m, sugar protons), 3.57 (3H, s, COOCH3).

Spectrophotometric studies of alkali and alkali earth metal ions complexes of mono amino derivative of calix[4]arene

The synthesis and complexive abilities of 5,11,17-tris(tert-butyl)-23 amino-25,26,27,28-tetra-propoxycalix[4]arene towards alkali cations Li+, Na+, K+, Rb+, Cs+ and alkali earth cations Mg2+, Ca2+, Sr2+ and Ba2+ in methanol-chloroform mixture have been evaluated at 25°C, using UV-Vis spectrophotometric techniques. The results showed that the ligand is capable to complex with all the cations by 1: 1 metal to ligand ratios. The selectivity presented considering the calculated formation constants are in the order Li+ > Na+ > K+ > Rb+ > Cs+ and Mg2+ > Ca2+ > Sr2+ > Ba2+ with the ligand.

Rapid analysis of free fatty acid composition in Brassica rapa L. and Brassica napus L. extracts by surface-assisted laser desorption/ionization time-of-flight mass spectrometry

A rapid and simple technique was developed to identify free fatty acids in mixtures by the surface-assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-TOF MS) using powdered carbon aerogels as a “matrix”. A 3% solution of a neutral surfactant Triton X-100 was used as a fixing agent of carbon particles as well as a suppressor of background peaks. The extraction of free fatty acids from Brassica napus (rape) and Brassica rapa (turnip) seeds was performed by using both methanol-chloroform mixture and supercritical fluid extraction techniques. The quantitative analysis of the seed composition was carried out to demonstrate the effectiveness of the developed method. The detection limits were determined using a mixture of ten standard fatty acids and were found to be 3 ng of each acid per spot.

Antimicrobial and cytotoxic activity of the stem and root bark extracts of Quassia undulata and isolated constituents

The stem and root barks of Quassia undulata were exhaustively extracted with methanol to give the methanolic extracts: MeQuS and MeQuR, respectively. The chloroform extracts were obtained and fractionated on an open column of silica gel by gradient elution with chloroform and chloroform–methanol mixtures. Purification of fractions afforded the isolation of four compounds, namely: 3′-(8-hydroxy,6-methoxy,2-oxo-2H-1-benzopyran-7-yloxy)-3′,4′-dihydro, 7′-methoxy,2H-1-benzopyran (eniotorin) (1) a new bisbenzopyran coumarin, 7-hydroxy-6-methoxy-2H-1-benzopyran-2-one (scopoletin) (2) (a coumarin); glaucarubinone (3) and 15-desacetylundulatone (4) (quassinoids). The antimicrobial activities of the extracts and isolated compounds were evaluated against five microbes, namely: Staphylococcus aureus; Eschericha coli; Pseudomonas aeruginosa; Klebsiella pneumonia and Candida albicans. The extracts exhibited a broad-spectrum activity against the tested microbes. Cytotoxic activities of the extracts and compounds isolated from them were also examined against brine shrimps, Artemia salina. The extracts and the compounds showed significant dose-dependent activities.

Rapid UPLC-MS/MS method for routine analysis of plasma pristanic, phytanic, and very long chain fatty acid markers of peroxisomal disorders

Quantification of pristanic acid, phytanic acid, and very long chain fatty acids (i.e., hexacosanoic, tetracosanoic, and docosanoic acids) in plasma is the primary method for investigateing a multitude of peroxisomal disorders (PDs). Typically based on GC-MS, existing methods are time-consuming and laborious. In this paper, we present a rapid and specific liquid chromatography tandem mass spectrometric method based on derivatization with 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole (DAABD-AE). Derivatization was undertaken to improve the poor mass spectrometric properties of these fatty acids. Analytes in plasma (20 mul) were hydrolyzed, extracted, and derivatized with DAABD-AE in approximately 2 h. Derivatives were separated on a reverse-phase column and detected by positive-ion electrospray ionization tandem mass spectrometry with a 5 min injection-to-injection time. Calibration plots were linear over ranges that cover physiological and pathological concentrations. Intraday (n = 12) and interday (n = 10) variations at low and high concentrations were less than 9.2%. Reference intervals in normal plasma (n = 250) were established for each compound and were in agreement with the literature. Using specimens from patients with established diagnosis (n = 20), various PDs were reliably detected. In conclusion, this method allows for the detection of at least nine PDs in a 5 min analytical run. Furthermore, this derivatization approach is potentially applicable to other disease markers carrying the carboxylic group.