Development of a bioanalytical method for the quantification of sinococuline in human plasma and its application in phase I clinical pharmacokinetic study

A selective, highly sensitive, precise, and novel bioanalytical method has been developed and validated to quantify sinococuline, an active constituent present in the phytopharmaceutical drug product containing Cocculus hirsutus plant extract, in vivo. Chromatographic separation was achieved on a Luna Omega Polar-C18 bonded analytical column maintained at 45°C. The isocratic mobile phase consisted of methanol and ammonium formate buffer (60:40, v/v) at acidic pH with a low flow rate of 0.250 mL/min. Detection was performed on an API 4000 mass spectrometer using electrospray ionization in positive polarity and multiple reaction monitoring mode to achieve a lower limit of quantification of 1.50 ng/mL. Excellent accuracy and precision were obtained after extracting the analyte from plasma samples using a chemical analogue as an internal standard in the absence of an isotope-labeled compound. The extraction efficacy was evidenced from recovery study, and the analyte was found to be stable in plasma. Validation study demonstrated linearity with coefficient of correlation, r ≥ 0.99, and minimal matrix effect. This bioanalytical method was successfully applied to evaluate pharmacokinetic parameters of sinococuline from a phase I clinical trial of an aqueous extract of C. hirsutus in healthy human volunteers.     

One-pot Synthesis,Crystal Structures and Antimicrobial Activities of Two New 1,4-Disubstituted 1,2,3-Triazole-4-Carboxylates

Two new 1,4-disubstituted 1,2,3-triazoles-4-carboxylates were synthesized via click reaction. Compound 1a was synthesized by the interaction of 6-nitro-tetrazolo[1.5-a]-pyridine with ethyl propynoate at room temperature in the presence of Cu(OAc)2 as a catalyst and THF as solvent. Compound 1b was also synthesized by the same manner except that tert-butyl propionate, instead of ethyl propynoate, was used. The compounds were characterized by IR, 1H-NMR, 13C-NMR and single-crystal X-ray diffraction analysis. Compound 1a(C10H9N5O4) crystallizes in the triclinic system, space group P1 with a = 5.0894(9), b = 8.9834(13), c = 13.089(2) ?, α= 83.041(7), β= 80.256(7), γ=87.296(8)°, V = 585.24(16)?3, Z = 2, Mr = 263.22, crystal size(mm) = 0.37 × 0.20 ×0.18,(I 2σ(I)) = 8557, 2493, 1229, Rint = 0.057. Compound 1b(C12H13N5O4) crystallizes in the monoclinic system, space group P21/c with a = 6.8854(5), b = 21.783(2), c = 9.3986(8) ?,β = 93.239(4)°, V = 1407.4(2)?3, Z = 4, Mr = 291.27, crystal size(mm) = 0.38 × 0.22 × 0.20,(I 2σ(I)) = 11842, 3172, 1866, Rint = 0.047. Antimicrobial assay results showed that the title compounds display excellent activities to different bacterial and fungal strains.     

On some properties of linear and nonlinear waves in pair-ion plasmas

It is pointed out that the observation of the electrostatic ion acoustic wave frequency can be a suitable check to determine whether the produced plasma is a pure pair-ion plasma or whether it comprises some concentration of electrons. A theoretical model for the pair-ion plasma dynamics is presented along with a new electrostatic mode which can exist only in such systems. It can become unstable in the presence of shear flow and it can give rise to vortex structures in the nonlinear regime. The possibility of shocks and solitons, due to nonlinear drift waves in a pair-ion plasma comprising electrons, is also discussed. The relevance of this investigation to both laboratory and astrophysical plasmas is pointed out.     

1,1‐Organoboration of bis(trimethylstannyl)ethyne with trimethylsilyl‐ and dimethylsilyl‐dialkylboryl‐substituted alkenes: organometallic‐substituted allenes

The reactions of bis(trimethylstannyl)ethyne, Me₃Sn–C?C–SnMe₃ ( 4 ), with trimethylsilyl‐ or dimethylsilyl‐dialkylboryl‐substituted alkenes 1 – 3 afford organometallic‐substituted allenes 5 , 6 and 8 , 9 in high yield. In the case of (E)‐2‐trimethylsilyl‐3‐diethylboryl‐2‐pentene ( 1) , a butadiene derivative 7 could be detected as an intermediate prior to rearrangement into the allene. All reactions were monitored by ²⁹Si and ¹¹⁹Sn NMR, and the products were characterized by an extensive NMR data set (¹H, ¹¹B, ¹³C, ²⁹Si, ¹¹⁹Sn NMR). Copyright © 2003 John Wiley & Sons, Ltd.     

Soft-laser scanning densitometry performed on a 35-mm slide illustrating SDS-PAGE separation of adenovirion polypeptides

Virion polypeptides (35 S), methionine-labeled and purified by CsCl gradient centrifugation, were separated by SDS-gel electrophoresis. Analysis of their band pattern was performed by scanning the images of the SDS-gels shown on a 35-mm slide. The densitometric tracings revealed the presence of 17 protein bands, although only 15 of them were visible to the naked eye. The high sensitivity and resolving capacities of the soft-laser scanning densitometer enabled us to detect trace amounts of protein bands separated in SDS-gels and to obtain a resolution compatible to that of electrophoresis. Fourfold electronic amplification of the densitometric tracings, produced by a computer, generated new information regarding the pattern of the electrophoregram. The facility to amplify peaks of importance is particularly advantageous when faint or overlapping protein bands revealed on a gel are assessed.     

Synthesis and structure of fluorotricyanomethane

Fluorotricyanomethane is prepared from potassium tricyanomethanide and perchlorylfluoride. The crystal structure of this compound (orthorhombic, space group P2₁2₁2₁, with a=6.270, b=6.734, c=11.776 Å) has been determined by X-ray analysis. The results are discussed in relation to those of (NC)₃C? X (X=? CH₃, ? Cl, ? Br or ? CN).     

Aqueous phenylacetic acid as a low-dose fluorescence dosimeter

Total fluorescence of aqueous phenylacetic acid system at neutral pH has been evaluated as low-dose -ray chemical dosimeter, using the fluorescence accessory of a UV-Vis spectrophotometer. The useful working range of the system is from 4 to 70 Gy. The postirradiation stability of the dosimeter response has been studied at different pH's, storage temperatures and light conditions. The post-irradiation stability of the dosimeter decreases with an increase in storage temperature. The stability is not affected in diffuse sunlight but it is very unstable in direct sunlight.     

A Study of the Uptake of Toluidine Blue O by Porphyromonas gingivalis and the Mechanism of Lethal Photosensitization

The purpose of the study was to determine the distribution of the photosensitizer toluidine blue O (TBO) within Porphyromonas gingivalis and the possible mechanism(s) involved in the lethal photosensitization of this organism. The distribution of TBO was determined by incubating P. gingivalis with tritiated TBO (³H-TBO) and fractionating the cells into outer membrane (OM), plasma membrane (PM), cytoplasmic proteins, other cytoplasmic constituents and DNA. The percentage of TBO in each of the fractions was found to be, 86.7, 5.4, 1.9, 5.7 and 0.3%, respectively. The involvement of cytotoxic species in the lethal photosensitization induced by light from a helium-neon (HeNe) laser and TBO was investigated by using deuterium oxide (D₂O), which prolongs the lifetime of singlet oxygen, and the free radical and singlet oxygen scavenger L-tryptophan. There were 9.0 log₁₀ and 2 log₁₀ reductions in the presence of D₂O and H₂O (saline solutions), respectively, at a light dose of 0.44 J (energy density = 0.22 J/cm²), suggesting the involvement of singlet oxygen. Decreased kills were attained in the presence of increasing concentrations of L-tryptophan. The effect of lethal photosensitization on whole cell proteins was determined by measuring tryptophan fluorescence, which decreased by 30% using 4.3 J (energy density = 4.3 J/ cm²) of light. Effects on the OM and PM proteins were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. There was evidence of change in the molecular masses of several PM proteins and OM proteins compared to controls. There was evidence of damage to the DNA obtained from irradiated cells. Scanning electron microscopic studies showed that there was coaggre-gation of P. gingivalis cells when sensitized and then exposed to laser light. These results suggest that lethal photosensitization of P. gingivalis may involve changes in OM and/or PM proteins and DNA damage mediated by singlet oxygen.