Development of a rapid resolution HPLC method for the separation and determination of 17 phenolic compounds in crude plant extracts

Rapid Resolution HPLC/DAD method, on a 1.8 μm, 4.6×50 mm column, was developed to enable a rapid separation of a mixture of 17 compounds, which consisted of hydroxybenzoic acids, hydroxycinnamic acids, flavones, flavonols, flavanone, flavonol-glycoside and antraquinone, in a single run, within 22 minutes. The developed method is precise, accurate and sensitive enough for simultaneous quantitative evaluation of major compounds in crude and hydrolyzed extracts of parsley, buckthorn, mint, caraway and birch. In order to overcome the inability to quantify all the phenolic compounds present in the samples caused by lack of external standards, HPLC approaches for the total phenolic content estimation based on sum of all integrated peak areas were made. These results were compared with the total phenolic content determined by Folin-Cioacalteu method. Although the correlation between the series of data was not significant (p<0.05), the difference between the results of total phenolic content obtained spectrophotometrically and by HPLC was not high in the case of parsley, buckthorn and mint extract. Regarding the obtained results, the HPLC approach could serve as an excellent tool for total phenolic content estimation, without the need for complete identification of the individual compounds.      

Effect of substituents on redox, spectroscopic and structural properties of conjugated diaryltetrazines--a combined experimental and theoretical study

Two series of new soluble conjugated compounds containing tetrazine central ring have been synthesized. The three-ring compounds have been synthesized by the reaction of aryl cyanide (where aryl = thienyl, alkylthienyl, phenyl or pyridyl) with hydrazine followed by oxidation of the intermediate product with diethyl azodicarboxylate. The five-ring compounds have been prepared using two pathways: (i) reaction of 5-cyano-2,2'-bithiophene (or its alkyl derivative) with hydrazine; (ii) via Suzuki or Stille coupling of 3,6-bis(5-bromo-2-thienyl)-1,2,4,5-tetrazine with a stannyl or boronate derivative of alkylthiophene. UV-vis spectroscopic properties of the synthesized compounds are strongly dependent on the nature of the aryl group, the position of the solubilizing substituent and the length of the molecule, showing the highest bathochromic shift (λ(max) > 440 nm) for five-ring compounds with alkyl groups attached to C(α) carbon in the terminal thienyl ring. An excellent linear correlation has been found for spectroscopically determined and theoretically calculated (TD-B3LYP/6-31G*) excitation energies. With the exception of dipyridyl derivative, the calculated lowest unoccupied molecular orbital (LUMO) level of the investigated molecules changes within a narrow range (from -2.63 to -2.41 eV), in line with the electrochemical data, which show a reversible reduction process with the redox potential varying from -1.23 V to -1.33 V (vs. Fc/Fc(+)). The electrochemically determined positions of the LUMO levels are consistently lower by 0.9 to 1.2 eV with respect to the calculated ones. All molecules readily crystallize. Single crystal studies of 3,6-bis(2,2'-bithien-5-yl)-1,2,4,5-tetrazine show that it crystallizes in a P2(1)/c space group whose structural arrangement is not very favorable to the charge carriers flow within the crystal. Powder diffraction studies of other derivatives have shown that their structural organization is sensitive to the position of the solubilizing substituent. In particular, the presence of alkyl groups attached to C(α) carbon in the terminal thienyl ring promotes the formation of a lamellar-type supramolecular organization.     

Syntheses and structures of the first terminal phosphanylphosphido complex of hafnium [Cp2Hf(Cl){η(1)-(Me3Si)P-P(NEt2)2}] and the first zirconocene-phosphanylphosphinidene dimer [Cp2Zr{μ(2)-P-P(NEt2)2}2ZrCp2

Reactions of (Et(2)N)(2)P-P(SiMe(3))Li with [Cp(2)MCl(2)] (M = Zr, Hf) in toluene or pentane yield the related terminal phosphanylphosphido complexes [Cp(2)M(Cl){η(1)-(Me(3)Si)P-P(NEt(2))(2)}]. The solid state structure of [Cp(2)Hf(Cl){η(1)-(Me(3)Si)P-P(NEt(2))(2)}] was established by single crystal X-ray diffraction. The reaction of (Et(2)N)(2)P-P(SiMe(3))Li with [Cp(2)ZrCl(2)] in THF or DME solutions leads to the formation of deep red crystals of the first neutral diamagnetic zirconocene-phosphanylphosphinidene dimer [Cp(2)Zr{μ(2)-P-P(NEt(2))(2)}(2)ZrCp(2)]. The molecular structure of this compound was confirmed by X-ray diffraction. The reactions of (R(2)N)(2)P-P(SiMe(3))Li with [CpZrCl(3)] yield the related tetraphosphetanes R(2)NP(μ(2)-PSiMe(3))(2)PNR(2), which apparently are formed as a result of a transfer of NR(2) groups from a P atom to the Zr atom.     

A Raman microspectroscopic study of organic inclusions in “watermelon” tourmaline from the Paprok mine (Nuristan, Afghanistan)

An exceptional mineral specimen, particularly interesting for collectors and researchers, is “watermelon” tourmaline, mined in the Paprok region in Nuristan, Afghanistan. A pale pink core (zone III) is surrounded by a thin, colorless zone (zone II), and a green outer zone (zone I). Differences between zones are clearly seen in the SEM picture, i.e., diversity in the size of constituent forms, and also the presence of inclusions of alien mineral phases which range from a few dozen to a few μm. In the line mapping experiment the Raman spectra were collected from the inner core towards the outer rim. There are observed typical for tourmaline (elbaite) groups of bands: at ca. 3600, 1100, 700, 400 and 250 cm⁻¹. Raman measurements determine the differences in the local chemical composition of particular zones. Interestingly, there is a new band emerging in the outer zone at ca. 840 cm⁻¹, and it could be due to the SiO₄ stretching vibrational modes predisposed by the Fe ion presence in the mineral. The organic inclusions are present in the whole volume of the investigated gem. Organic inclusion content appears to be versatile. Raman spectra confirm the presence of hydrogen sulphide (2610 cm⁻¹), methane (2912 cm⁻¹), ethane (2950 cm⁻¹), and propane (2895 cm⁻¹). In the corresponding bending vibrations region apart from CH₂, CH₃ vibrations and C–O stretching (CO₂) at (1280, 1390 cm⁻¹), characteristic bands are also observed at ca. 1340, 1600 and 1660 cm⁻¹. The last ones possibly mark the evolution of graphitization process occurring from fluid saturated inclusions within condensed ring aromatics (1660 cm⁻¹ band), towards slightly more organized, although still amorphous forms of carbonaceous matter (1340 and 1600 cm⁻¹).     

Evaluation of the potential of surface enhancement Raman spectroscopy for detection of tricyclic psychotropic drugs. Case studies on imipramine and its metabolite

The potential use of surface Raman enhanced spectroscopy (SERS) for confirmatory identification and the semi-quantitative analysis of selected tricyclic antidepressants (TCAs) is examined utilizing a conventional silver colloid. Raman and SERS spectra of aqueous solutions of imipramine (Imi) and its metabolite, desipramine (Des), were recorded as the function of concentration using NIR excitation. A good linear correlation is observed for the dependence of the SERS signal at 684 cm(-1) (R(2) = 0.9997) on Imi concentration over the range of 0.75-7.5 μM. The limit of detection of imipramine in the silver colloidal solution is 0.98 μM. SERS spectra of Imi and Des were also recorded for blood plasma samples without prior purification as well as after the use of standard solid phase extraction. All spectra show the characteristic spectral profile of the molecules and moreover, stronger signal enhancement is observed for Imi in the "raw" samples as opposed to Imi extracted from a biological matrix.     

Pyrolysis-gas chromatography-mass spectrometry for studying N-vinyl-2-pyrrolidone-co-vinyl acetate copolymers and their dissolution behaviour

Knowledge on the solubility behaviour and dissolution rate of speciality and commodity polymers is very important for the use of such materials in high-tech applications. We have developed methods for the quantification and characterization of dissolved copolymers of N-vinyl-2-pyrrolidone (VP) and vinyl acetate (VA) during dissolution in water. The methods are based on pyrolysis (Py) performed in a programmed-temperature vaporization injector with subsequent identification and quantification of the components in the pyrolysate using capillary gas chromatography–mass spectrometry (GC–MS). By injecting large volumes and applying cryo-focussing at the top of the column, low detection limits could be achieved.The monomer ratio was found to have the greatest effect on the dissolution rate of the PVP-co-VA copolymers. The material with the highest amount of VA (50%) dissolves significantly slower than the other grades. Size-exclusion chromatography (SEC) and Py–GC–MS were used to measure molecular weights and average chemical compositions, respectively. Combined off-line SEC//Py–GC–MS was used to determine the copolymer composition (VP/VA ratio), as a function of the molecular weight for the pure polymers. In the dissolution experiments, a constant VP/VA ratio across the dissolution curve was observed for all copolymers analysed. This suggests a random distribution of the two monomers over the molecules.     

Black painted pottery, Kildehuse II, Odense County, Denmark

This work aimed at characterization of a black layer covering a Bronze Age (period VI) pot surface. To solve research problems plane polarized light microscopy (PLM), scanning microscopy with X-ray microanalyser (SEM/EDS), Raman and FTIR microspectroscopies were applied. Observation of the black layer under the PLM suggests that we deal with an opaque, isotropic layer, purposely put on leather-hard, burnished surface of a pot. No traces of organic substances that might have modified a pot surface before painting were detected. The black layer coats the pot with a continuous 0.2-0.4 μm thick layer. The black layer must have been applied while warm or hot. It is recognized that the black layer is a true painting layer. Spectroscopic analyses (Raman and FTIR) point that birch tar or a birch-pine tar had been applied. Carbon black should be definitely excluded. The analyzed organic substance is structurally ordered in various degrees, due to varying temperature influence. Some Raman spectra reveal, in second-order region of the spectrum (>2000 cm(-1)), the graphite presence. Scarce points reveal the presence of inorganic admixtures: clay minerals, feldspars and quartz. The next question is: Is it an organic paint or a carbon-based, "mixed" paint? Inorganic admixtures are only casual. Relatively high concentration of alumina is expected to have appeared due to aluminum mobility in acidic environment and this was provided by organic tar/pitch substances. Phosphorus (usually with calcium) mostly concentrates together with clay minerals. Thus, hypothesis on bone powder or ash addition to tar/pitch should be excluded. Some elements may originate not only from inorganic substances but also from wood ashes.     

Determination of lipophilicity of γ‐butyrolactone derivatives with anticonvulsant and analgesic activity using micellar electrokinetic chromatography

The lipophilicity of a library of 30 derivatives of dihydrofuran‐2(3H)‐one (γ‐butyrolactone) was determined by MEKC. Calibration curve prepared for ten reference drugs enabled to calculate partition coefficient (log P) for novel compounds. The results of MEKC analysis were compared with lipophilicity coefficients determined by RP‐TLC (R_M0) and computational (Mlog P, Clog P) methods. Good correlation was observed between the results obtained by both experimental methods: the MEKC parameters log k and relative lipophilicity R_MO. The relationship between determined log P values and results of the computational prediction was weaker. Analysis of the relationship between lipophilicity and anticonvulsant activity showed statistically significant differences between mean values of log P coefficients for group of active (2.18) and inactive (1.51) compounds in the maximal electroshock test.     

Preparation of aluminium ammonium calcium phosphates using microwave radiation

Microwave radiation was used in the acquisition of aluminium ammonium calcium phosphates. The substrates such as CaCO₃, H₃PO₄, aqueous ammonia were reagent grade, whereas Al(OH)₃ was prepared afresh. The influence of process parameters (pH 6 ± 2, molar ratios of Al³⁺: Ca²⁺: PO ₄ ^3? in the substrates, respectively 0.31: 0.62: 1; 0.5: 0.5: 1; 0.72: 0.36: 1) on the phase composition and the product properties was determined. Statistical software STATISTICA 10 was used for planning and evaluation of the experiments. The process parameters making it possible to acquire the material with the anticipated physicochemical properties were determined based on statistical evaluation of the planned research by the plan fractional factorial design at three levels 3^(k?p). The phase composition of the samples was studied using XRD analysis. The specific surface area was calculated using the BET method and the particle size was determined by LSM. Materials with a molar ratio of Al³⁺: Ca²⁺ and Al³⁺: NH ₄ ⁺ in the range of 0.07–0.76 and 0.75–3.4, respectively, with an absorption oil number of 36–56 g per 100 g, S _BET within 8.2–73 m² g^?1, and particle size in the range of 156–252 nm were obtained.     

Age-Related Carbonylation of Fibrocartilage Structural Proteins Drives Tissue Degenerative Modification

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