A GC-SIM-MS Method for the Determination of Butylidenephthalide in Rat Plasma and Tissue: Application to the Pharmacokinetic and Tissue Distribution Study

Abstract

Butylidenephthalide is one of the major active components isolated from Rhizoma Chuanxiong. This paper describes a simple, rapid, specific and sensitive method for the quantification of butylidenephthalide in rat plasma and tissue distribution using a liquid-liquid extraction procedure followed by capillary gas chromatography-selected ion monitoring mode-mass spectrometry (GC-SIM-MS) analysis. The calibration curves were linear over the concentration ranging from 0.02–10.0 µg/mL (r > 0.99) for plasma samples and 0.18–7.25 µg/g (r > 0.99) for the tissue samples. The limit of quantification (LOQ) was 1.0 ng/mL or 1.0 ng/g (ten times signal/noise ratio). Within- and between-day precisions expressed as the relative standard deviation (RSD) for the method were 2.39–2.98% and 2.97–4.26%, respectively. The methods of recovery for all samples were greater than 80% at the low, medium, and high concentrations. The method has been successfully applied to a pharmacokinetics study in rats after an oral administration of Butylidenephthalide with a dose of 20.0 mg/kg. The main pharmacokinetic parameters obtained were T _max  = (0.22 ± 0.06) h, C _max = (3 ± 1) µg/mL, AUC = (32 ± 6) h?µg/mL, and K _a  = (8.5 ± 0.8)/h. The results showed that the butylidenephthalide was easily absorbed. The concentrations of butylidenephthalide in rat kidney, lung, heart, and cerebellum were higher than those in other organs. To determine free fraction in serum, samples were filtered using ultrafiltration membranes with a molecular weight cut-off of 10,000 Da and extracted using liquid-liquid extraction. The extracts were evaporated and analyzed by GC-MS. The protein binding in rat plasma, human plasma, and human serum albumin were 83 ± 4%, 94 ± 3%, and 89 ± 3%, respectively.     

Two new sesquiterpene lactones from the fruits of Illicium jiadifengpi

Two new seco-prezizaane-type sesquiterpenes, 3,4-dehydroneomajucin (1) and 1,2,3,4-tetradehydroneomajucin (2), were isolated from the fruits of Illicium jiadifengpi. The structure of these compounds was determined using 1D and 2D NMR and ESI-MS. The isolates were evaluated for their anti-hepatitis B virus activities on the Hep G2.2.15 cell line. The inhibitory rates of compounds 1 and 2 on the HBeAg and HBsAg expression were 30.08 ± 3.09% and 11.43 ± 1.92% at a concentrations of 68.00 μM and 7.88 ± 1.21% and 16.96 ± 4.24% at a concentration of 68.50 μM, respectively.     

Analysis of Plasma Angiotensins by Reversed Phase HPLC and Radioimmunoassay

Abstract

The analysis of biologically active angiotensin peptides in blood plasma by high performance liquid chromatography in a weakly non-polar reversed phase (C₂) chromatographic system combined with quantification of chromatographically isolated peptides by radioimmunoassay has been developed. This system is able to resolve each of seven closely-related peptides of the angiotensin group. The chromatographic system was applied to plasma samples which have been prepared for chromatographic analysis by C₁₈ cartridge extraction. Samples were reconstituted in HPLC solvent prior to injection into the HPLC system. Separated angiotensin were collected by fraction collector and the volatile components of the solvent system were blown off under an air stream. The content of several of the various angiotensin peptides in the fractions was then determined by radioimmunoassay using an appropriate antiserum. Antiserum to angiotensin II (octapeptide) was used to quantify the biologically active components angiotensin II, angiotensin III (hepta-peptide) and C-terminal hexapeptide. Recovery of angiotensin II in the C₁₈ cartridge extraction has been assessed at 85.0 ± 0.9% (mean ± SEM) using I¹²⁵-labelled angiotensin II, and 82.2 ± 4.45% using synthetic unlabelled angiotensin II. Recovery of standard preparations of angiotensin II in the HPLC system have been estimated at 67.5 ± 6.08%. The application of this technique to evaluating some components of the angiotensin in normal plasma is presented.     

Determination of the Antineoplastic Agent Tiazofurin in Plasma by High Performance Liquid Chromatography

Abstract

A simple and sensitive sample clean up procedure and a reverse phase high performance liquid chromatographic assay for tiazofurin in dog plasma has been developed.

Thymidine was used as the internal standard and the elution solvent was 0.16 M acetic acid. The retention times of tiazofurin and thymidine at an elution rate of 2.0 ml/minute were 8.7 minutes and 16 minutes respectively. The assay sensitivity was 1.0 μG/ml. The within run coefficient of variation of 2.0 μG/ml and 20 μG/ml plasma samples were ±7.49% and ±3.37% respectively and the between run coefficient of variation of 2.0 μG/ml and 20.0 μG/ml plasma samples were ±10.46% and ±5.92% respectively.     

Elemental Analysis and Metal Intake of Romanian Vegetables

ABSTRACT

Inductively coupled plasma-mass spectrometry multielemental profiling was performed to determine eight elements in 54 vegetable samples. The range of concentrations of Fe and Ni was 1.91?±?0.09 to 443.31?±?22.32?mg/kg and 0.05?±?0.004 to 16.36?±?0.36?mg/kg, respectively. The concentrations of Pb and Cd were up to 1.56?±?0.04 and 0.78?±?0.04?mg/kg, respectively. Some samples exceeded the maximum admissible limits imposed by national and international regulations. The mean concentrations of Mn and Cu were between 20.93?±?0.43 and 13.98?±?0.30?mg/kg in tomatoes and 20.60?±?0.53 and 11.25?±?0.22?mg/kg in peppers. For the evaluation of health risk related to the consumption of vegetables, the daily intake rate, target hazard quotient (THQ), and hazard index were evaluated. The daily intakes were low and correspond to 2 to 14% of the tolerable daily values established by legislation. The results showed that the THQ and hazard index for the metals were lower than 1, indicating that the vegetable ingestion does not represent a significant risk for consumer health. Analysis of variance revealed that Zn may be used as a species-specific marker (p?=?0.026). Principal component analysis extracted three main components, explaining a total variance of 72.49% and grouped all determined elements.     

Determination of imidacloprid in paddy water and soil by liquid chromatography electrospray ionization-tandem mass spectrometry

A method for the determination of imidacloprid in paddy water and soil was developed using liquid chromatography electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS). Separation of imidacloprid was carried out on a Shimadzu C18 column (150 mm × 4.6 mm, 4.6 μm) with an acetonitrile-water (50: 50, v/v) mobile phase containing 0.1% of acetic acid. The flow rate was 0.3 mL/min in isocratic mode. The product ion at 209 m/z was selected for quantification in multiple-reaction monitoring scan mode. Imidacloprid residues in soil were extracted by a solid-liquid extraction method with acetonitrile. Water samples were filtered and directly injected for analysis without extraction. Detection limits of 0.5 μg/kg and 0.3 μg/L were achieved for soil and water samples, respectively. The method had recoveries of 90 ± 2% (n = 4) for soil samples and 100 ± 2% (n = 4) for water samples. A linear relationship was observed throughout the investigated range of concentrations (1–200 μg/L), with the correlation coefficients ranging from 0.999 to 1.000.     

A Simple and Fast Method for Manganese Determination in Antihypertensive Drugs by Slurry Sampling Graphite Furnace Atomic Absorption Spectrometry

Abstract

A simple and rapid procedure for the determination of manganese in anti‐hypertensive drugs is proposed. The samples were treated with dilute nitric acid (1.2% v/v) with stirring using a vortex stirrer to yield a slurry. To determine the best conditions for analysis by graphite furnace atomic absorption spectrometry (GF AAS), a planning factorial was initially employed that demonstrated that the non‐use of chemical modifiers and the use of lower pyrolysis temperatures were more appropriate. Next, the pyrolysis and atomization temperature curves were obtained. The best pyrolysis and atomization temperatures encountered were 500 and 2200°C, respectively. Calibration was performed by matrix matching. The characteristic mass was 0.70±0.14 pg (the recommended mass was 0.60 pg); the limits of detection and quantification were 0.23 and 0.77 µg l^?1, respectively. The precision obtained in the intra‐ and inter‐assay studies of the drug spiked with 0.5, 1.0 and 1.5 µg l^?1 of Mn did not exceed 11% (n=7). Recovery studies of the drug spiked with three levels (n=7) of Mn yielded results between 101.0±4.5 and 116.3±9.7%. The results of an analysis of a certified urine sample (two levels of Mn) agreed at a 95% level of confidence. Forty‐eight antihypertensive drug samples were analyzed, and the results varied from 2.9 ng to 1.9 µg of Mn per capsule^?1.     

Alkaline hydrolysis of ethiofencarb: Kinetic study and mechanism degradation

The present paper deals with the hydrolysis of ethiofencarb [2‐ethylthiomethyl(phenyl)‐N‐methylcarbamate] in alkaline solution. The reaction kinetics has been investigated using spectrophotometric and liquid chromatographic techniques. The rate constants were determined following a proposed first‐order kinetic model. The positive activation entropy Δ S^≠ = +100.07 J mol^?1 K^?1 and the absence of general basic catalysis indicated an E1cB hydrolytic mechanism, involving the formation of methyl isocyanate. This result was confirmed by the fact that ethiofencarb fits well into Brönsted and Hammett lines, obtained for a series of substituted N‐methylcarbamate whose decomposition in aqueous media was established to follow an E1cB mechanism. © 2012 Wiley Periodicals, Inc. Int J Chem Kinet 45: 118–124, 2013     

Liquid Chromatographic Determination of Dimetridazole and Ronidazole in Feeds

Abstract

A simple high performance liquid chromatographic (HPLC) procedure for the simultaneous determination of dimetridazole and ronidazole in turkey feeds is described. The drugs are extracted from feeds by carbon-tetrachloride/dimethylformamide (80:20) at 60°C during 30 mn and the extract is subjected to a partition by water. After centrifugation the eluate is chromatographed on a reverse phase column with ultaviolet detection at 316 nm. Recoveries from samples fortified at levels 2.02 to 7.07 ppm for ronidazole and 2.01 to 7.03 for dimetridazole were 99,6% ± 1,4 and 95,3% ± 1,8 (mean ± standard deviation), respectively.     

New jacaranone glucoside from Jacaranda oxyphylla leaves

Jacaranda oxyphylla Cham. is popularly known as ‘caroba-de-São-Paulo’ and it is used in traditional medicine for microbial infections. A new phytoquinoid (α/β-glucoside-4-phenylacetate-6-(1-hydroxy-4-oxo-2,5-cyclohexadiene-1-acetate) (1) was isolated from J. oxyphylla leaves, together with three known compounds: quercetin-3-O-β-d-galactoside (2), verbascoside (3) and polystyrene (4). Their chemical structures were elucidated using spectroscopic techniques and by comparison with the related known compounds. In addition, it was found a pronounced acetylcholinesterase inhibitory activity for the quinoid 1 (100.0 ± 0.8%) and phenolic compounds 2 and 3 (99.9 ± 0.7 and 99.3 ± 0.5%, respectively), if compared to the standard eserine (92.7 ± 0.4%), that was analysed by a microplate spectrophotometer.     

Simultaneous Determination of Cadmium,Copper, Lead and Zinc in Amino Acid Parenteral Nutrition Solutions by Anodic Stripping Voltammetry and Sample Digestion by UV Irradiation

Abstract

Irradiation with ultraviolet (UV) energy was investigated to assay cadmium, copper, lead, and zinc by anodic stripping voltammetry (ASV) in amino acid parenteral nutrition (PN) solutions. Sample digestion by UV irradiation showed the best performances to liberate the metals from the samples (metal recoveries between 90% and 102%) in comparison with classical oxidative wet digestion methods. The best UV digestion condition was obtained with 1:10 diluted PN samples irradiated during 10 h at 90±3°C with the addition of one aliquot of 50 µL concentrated H₂SO₄ and repeated additions of 50 µL 30% (v/v) H₂O₂ at each 60 min irradiation interval. By using the UV digestion procedure cadmium, copper, lead, and zinc were simultaneously assayed in commercial amino acid PN solutions by ASV. The metal concentrations ranged between 1.3 to 4.4 for cadmium, 2.9 to 40.8 for copper, 4.4 to 16.8 for lead, and 1.4 to 208.5 for zinc. The ASV method correlated well with atomic absorption spectrometry measurements to assay the investigated analytes in amino acid PN samples after the UV digestion.     

Spectrophotometric determination of ethiofencarb in waters by reaction with p-aminophenol

Summary An automatized procedure has been developed for the spectrophotometric determination of ethiofencarb in water by reaction with p-aminophenol after alkaline hydrolysis to obtain the corresponding phenol sulphone. The hydrolyzed samples are continuously introduced into different manifolds at the same time as 300 mg/l p-aminophenol, 0.004 mol/l KIO₄ and 0.04 mol/l NaOH solutions. The absorbance is measured at 638 nm after a reaction time of 6 min in stop flow. This absorbance band corresponds to the indo dye obtained from the reaction between the phenol sulphone of ethiofencarb and the quinoneimine form of the p-aminophenol and it permits a matrix-free spectrophotometric determination of ethiofencarb with a limit of detection of 33 g/l. The recovery of ethiofencarb in different water matrices varies from 71 to 126%.     

Determination of acetylcholinesterase and α-glucosidase inhibition by electrophoretically-mediated microanalysis and phenolic profile by HPLC-ESI-MS/MS of fruit juices from Brazilian Myrtaceae Plinia cauliflora (Mart.) Kausel and Eugenia uniflora L.

Abstract

Alzheimer’s disease and diabetes mellitus are contemporary diseases of great concern. Phenolic compounds are linked to several health benefits and could lead to novel strategies to combat these ailments. The objective of this study was to evaluate by electrophoretically-mediated microanalysis the potential inhibitory activity of the fruit juices from Plinia cauliflora (“jaboticaba”) and Eugenia uniflora (“pitanga”) toward acetylcholinesterase (AChE) and α-glucosidase, target enzymes in strategies for the treatment of these diseases. The phenolic profiles of the samples were also investigated. Jaboticaba and pitanga juices inhibited 85.90?±?1.73 and 52.67?±?1.24% of AChE activity at 5?mg mL^?1, and 57.91?±?2.60 and 69.47?±?2.89% of α-glucosidase activity at 1?mg mL^?1, respectively. Total phenolic content of the juices were 303.54?±?28.28 and 367.00?±?11.42 mgGA L^?1, respectively. The observed inhibitory activity can be explained, at least in part, by the presence of the phenolic compounds.     

Synthesis,EGFR-TK inhibition and anticancer activity of new quinoxaline derivatives

Abstract

Ethyl 4-substituted-3-oxo-quinoxaline-2-carboxylates 3–5 were obtained via alkylation of ethyl 3-oxo-3,4-dihydroquinoxaline-2-carboxylate (1). Compound 1 was heterocyclized using hydrazines, ethylenediamine, and ethanolamine to give pyrazoloquinoxalines 6, 7, diazepinoquinoxaline 8, and oxazepinoquinoxaline 10. The quinoxaline-2-carboxamides 9, 11, 12 were prepared via condensation of compound 1 with different amines. Compound 1 was thiated using Lawesson’s reagent affording quinoxaline-3-thione 13, in fair yield. In addition, the reaction of 4-methyl-3-oxoquinoxaline 3 with some binucleophiles led to a series of new oxoquinoxaline derivatives 14–18. The molecular structure of compounds 1, 3, and 9 was confirmed by X-ray crystallography.

The anti-proliferative activity showed that among all the tested compounds, compounds 3, (IC₅₀ 2.51?±?3.0, 4.22?±?1.6 and 2.27?±?1.9?µM), 11 (IC₅₀ 1.32?±?2.61, 1.41?±?1.23 and 1.18?±?1.91?µM) and 17 (IC₅₀ 1.72?±?1.32, 1.85?±?0.94 and 1.92?±?4.83?µM) showed noteworthy anti-proliferative effects against the three cancer cell lines, HCT116, HePG2 and MCF7, respectively, compared to the reference drugs doxorubicin (IC₅₀ 1.41?±?0.58, 0.90?±?0.62 and 1.01?±?3.02?µM) and erlotinib (IC₅₀ 1.63?±?0.81, 1.57?±?0.62 and 1.49?±?0.54?µM). Compounds 3 (0.899?nM), 11 (0.508?nM) and 17 (0.807) showed strong EGFR inhibitory activity compared to Erlotinib (0.439?nM) and these results are in agreement with the docking study. These results suggest that compounds could probably be promising anticancer agents with EGFR inhibitory activity.     

Determination of Major and Minor Elements in Granite by Inductively Coupled Plasma and X-ray Fluorescence Spectrometry

Abstract

The development and evaluation of an analytical method for the determination of major and minor elements in granite by ICP-AES and XRF are described. The method applies a standard addition method to generate calibration standards for XRF measurements by ICP-AES from samples to be analyzed. Reliable results were obtained by using simple versions of ICP and XRF, and combined with the pressed-powder pellets for sample preparation. Least squares data for parameters measured by this method vs. reported value for a granite standard reference material gave a slope of 1.002 ± 0.005 and an intercept of -0.029 ± 0.07. The coefficients of variation vary between 0.53% for the largest oxide and 8.3% for the lowest oxide.     

Pervaporation as a Separation Technique for Direct Determination of Volatile Selenium Species by Atomic Fluorescence Spectrometry

Abstract

This paper reports for the first time a suitable way to determine methylated selenium compounds using the new approach of pervaporation coupled to atomic fluorescence spectrcmetry (PV-AFS).

The method developed allows direct extraction, separation, preconcentration and determination of dimethylselenium (DMSe) and dimethyldiselenium (DMDSe) from slurry samples. Under the optimum conditions, the detection limits (LODs) were found to be 0.66 ng and 0.39 ng for DMSe and DMDSe, respectively, the precision being about 6–9 % for 10 ng mL as selenium concentration. The linearity ranges were from the LOD to 0.7 μg mL^?1 for DMSe and from the LOD to 0.4 μg mL^?1 for DMDSe (as Se). The pervaporation efficiencies were 55 ± 1 % and 85 ± 5 % for DMSe and DMDSe, respectively. The proposed method was successfully applied to determine methylated selenium species in sewage sludge, garlic and oyster samples. The concentrations found were from 0.07 to 1.42 μg g^?1.

As no certified reference materials are available for these analytes, validation was carried out by recovery studies in these matrices, and the results showed that the proposed method performed satisfactorily.     

Anti-glycating and anti-oxidant compounds from traditionally used anti-diabetic plant Geigeria alata(DC) Oliv. & Hiern.

Abstract

A new sesquiterpene lactone geigerianoloide (1) and four known flavonoids axillarin (2), quercetin (3), 3-methoxy-5,7,3',4'-tetrahydroxy-flavone (4) and hispidulin (5) were isolated from Geigeria alata (DC) Oliv. & Hiern. (Asteraceae). Structures were deduced using ¹H- and ¹³C- NMR spectroscopy, mass spectrometry, while the structure of compound 1 was also deduced using X-ray crystallography technique.

Geigeria alata is traditionally used for diabetes, therefore compounds were tested for anti-glycation activity, in which compounds 2 and 3 showed potent activities (IC₅₀ values of 246.97?±?0.83 and 262.37?±?0.22 µM, respectively) compared to IC₅₀ value 294.50?±?1.5 µM of rutin. Moreover, compound 4 exhibited a comparable activity to rutin (IC₅₀?=?293.28?±?1.34 µM). Compound 5 showed a weak activity.

Compounds 2, 3, and 4 exhibited potent DPPH radical scavenging activity (IC₅₀?=?0.1?±?0.00, 0.13?±?0.00 and 0.15?±?0.01 µM, respectively). Compounds 2, 3, and 4 demonstrated significant superoxide anion scavenging activity with IC₅₀ values of 0.14?±?0.001, 0.17?±?0.00, and 0.11?±?0.006 µM, respectively.     

Determination of Low Molecular Weight Carboxylic Acids in Water by HPLC with Conductivity Detection

Abstract

Low molecular weight carboxylic acids (C₂-C₈) were measured in aqueous samples using reverse phase HPLC coupled with conductivity detection. The aqueous samples are adjusted to pH 2 with H₂SO₄ and extracted with diethyl ether. The ether is allowed to evaporate over distilled water. The acid concentrate is analysed using a Zorbax-ODS column (25 mm × 4.6 mm). The method is simple, fast, and because of the selectivity of the conductivity detector, is relatively free from interferences. Average precision is estimated to be approximately ± 13% for the overall method.     

Selective Determination of Harmol by Room-Temperature Phosphorimetry: A Comparative Performance with Micellar Electrokinetic Capillary Chromatography

Abstract

Selective determination of harmol in the presence of other β-carboline alkaloids without the need for previous separation of components was achieved by Solid Surface Room-Temperature Phosphorimetry (SSRTP) using HgCl₂. Detection of harmol at concentrations as low as 5.2 × 10^?7 mol L^?1 can be made in urine samples. Recovery of 100±12% was achieved. The analytical performance of SSRTP was compared to Micellar Electrokinetic Capillary Chromatography (MECC).     

Dynamic Vapour Generation and Head Space Determination of m-XYLENE for Pharmacokinetic Studies

Abstract

A custom designed vapour generation-exposure chamber assembly and head space analysis method were evaluated for use in pharmacokinetic investigations. The predicted m-xylene concentration was within 1% of the average value measured by gas chromatography-flame ionization detection (GC-FID) in the glass exposure chamber during controlled vapourization of m-xylene. The concentration variability was ± 13% and 4% respectively, with and without rats in the chamber. Blood and tissue samples from male, Sprague Dawley rats exposed to 1100 ppm of xylene, were analyzed by means of a static head space (HS)-GC analysis method. The average m-xylene level in the blood of exposed animals was ca. 5000 ng/mL. m-Xylene was detected in brain, kidney, skin, fat and liver. The analytical precision for duplicate samples was < ± 5% for all tissues, except liver and skin. The feasibility of detection of m-xylene in unhomogenized tissue by HS-GC analysis was demonstrated.