Simultaneous analysis of mitotane and its main metabolites in human blood and urine samples by SPE-HPLC technique

Adrenocortical carcinoma (ACC) is a rare malignancy with an incompletely understood pathogenesis and a poor prognosis. The adrenalytic activity of mitotane has made it the most important single drug in the treatment of ACC. Unfortunately, the exact mechanism of mitotane action is still unknown. It is believed that mitotane belongs to the class of drugs that require metabolic transformation by cytochrome P450 for therapeutic action; therefore determination of plasma levels of not only mitotane but also its metabolites would help in carrying out the treatment. The objective of this work was to develop and validate an SPE‐HPLC method for simultaneous determination of mitotane and its metabolites in different biological fluids. The sample preparation consisted of a solid‐phase extraction on a Discovery DSC₁₈ cartridge, while analysis of extracts was performed on a Symmetry C₁₈ column. The usefulness of the proposed method was confirmed by analysis of plasma, red cell and urine samples from patient chronically treated with 1.5 g of mitotane. The patient involved in this study had a high plasma concentration of mitotane and none of the investigated metabolites were found. In order to investigate whether the polymorphism of CYP2C9 and CYP2C19 enzymes could be related to the metabolism of mitotane, RT‐PCR analysis was performed. Copyright © 2012 John Wiley & Sons, Ltd.     

Selectivity of purine alkylation by a quinone methide. Kinetic or thermodynamic control?

The alkylation reaction of 9-methyladenine and 9-methylguanine (as prototype substrates of deoxy-adenosine and -guanosine), by the parent o-quinone methide (o-QM), has been investigated in the gas phase and in aqueous solution, using density functional theory at the B3LYP/6-311+G(d,p) level. The effect of the medium on the reactivity, and on the stability of the resulting adducts, has been investigated by using the C-PCM solvation model to assess which adduct arises from the kinetically favorable path, or from an equilibrating process. The calculations indicate that the most nucleophilic site of the methyl-substituted nucleobases in the gas phase is the guanine oxygen atom (O(6)) (DeltaG()(gas) = 5.6 kcal mol(-)(1)), followed by the adenine N1 (DeltaG)(gas) = 10.3 kcal mol(-)(1)), while other centers exhibit a substantially lower nucleophilicity. The bulk effect of water as a solvent is the dramatic reduction of the nucleophilicity of both 9-methyladenine N1 (DeltaG)(solv) = 14.5 kcal mol(-)(1)) and 9-methylguanine O(6) (DeltaG)(solv) = 17.0 kcal mol(-)(1)). As a result there is a reversal of the nucleophilicity order of the purine bases. While O(6) and N7 nucleophilic centers of 9-methylguanine compete almost on the same footing, the reactivity gap between N1 and N7 of 9-methyladenine in solution is highly reduced. Regarding product stability, calculations predict that only two of the adducts of o-QM with 9-methyladenine, those at NH(2) and N1 positions, are lower in energy than reactants, both in the gas phase and in water. However, the adduct at N1 can easily dissociate in water. The adducts arising from the covalent modification of 9-methylguanine are largely more stable than reactants in the gas phase, but their stability is markedly reduced in water. In particular, the oxygen alkylation adduct becomes slightly unstable in water (DeltaG(solv) = +1.4 kcal mol(-)(1)), and the N7 alkylation product remains only moderately more stable than free reactants (DeltaG(solv) = -2.8 kcal mol(-)(1)). Our data show that site alkylations at the adenine N1 and the guanine O(6) and N7 in water are the result of kinetically controlled processes and that the selective modification of the exo-amino groups of guanine N2 and adenine N6 are generated by thermodynamic equilibrations. The ability of o-QM to form several metastable adducts with purine nucleobases (at guanine N7 and O(2), and adenine N1) in water suggests that the above adducts may act as o-QM carriers.     

LC–MS methods combination for identification and quantification of trans-sinapoylquinic acid regioisomers in green coffee

The present study aims to both identify and quantify trans-sinapoylquinic acid (SiQA) regioisomers in green coffee by combined UHPLC-ESI-QqTOF-MS/MS and UHPLC-ESI-QqQ-MS/MS methods. Among the various mono-acyl chlorogenic acids found in green coffee, SiQA regioisomers are the least studied despite having been indicated as unique phytochemical markers of Coffea canephora (known as Robusta). The lack of commercially available authentic standards has been bypassed by resorting to the advantages offered by high-resolution LC–MS as far as the identification is concerned. SiQA regioisomers have been identified in several samples of Robusta and Coffea arabica (known as Arabica) commercial lots from different geographical origin and, for the first time, in different samples of coffee wild species (Coffea liberica and Coffea pseudozanguebariae). Quantification (total SiQA ranging from 3 to 5 mg/100 g) let to reconsider these chlorogenic acids as unique phytochemical markers of Robusta being present in the same quantity and distribution in C. liberica as well. Gardeniae Fructus samples (fruits of Gardenia jasminoides) have additionally been characterized as this matrix is recognized as one of the few naturally occurring SiQA sources. The SiQA regioisomer content (total SiQA about 80 mg/100 mg) fully supports the proposal to use this matrix as a surrogate standard for further studies.     

Effect of Cold Atmospheric Pressure Plasma on Maize Seeds: Enhancement of Seedlings Growth and Surface Microorganisms Inactivation

Cold atmospheric pressure ambient air plasma generated by Diffuse Coplanar Surface Barrier Discharge (DCSBD) was investigated for inhibition of native microbiota and potentially dangerous pathogens (Aspergillus flavus, Alternaria alternata and Fusarium culmorum) on the maize surface. Moreover, the improvement of germination and growth parameters of maize seeds was evaluated. Maize (Zea mays L.; cv. Ronaldinio), one of the most important cultivated crops worldwide, was selected as the research material. Electrical measurements confirmed the high volume power density (80 W cm^?3) of DCSBD plasma. Non-equilibrium plasma state evaluated using optical emission spectroscopy showed values of vibrational and rotational temperature (2700?±?300) K and (370?±?75) K, respectively. Changes on the plasma treated seeds surface were studied by water contact angle measurement, scanning electron microscope analysis and Fourier transform infrared spectroscopy. A complete devitalisation of native microbiota on the surface of seeds was observed after a short treatment time of 60 s (bacteria) and 180 s (filamentous fungi). The plasma treatment efficiency of artificially contaminated maize seeds was estimated as a reduction of 3.79 log (CFU/g) in F. culmorum after a 60-s plasma treatment, 4.21 log (CFU/g) in A. flavus and 3.22 log (CFU/g) in A. alternata after a 300-s plasma treatment. Moreover, the obtained results show an increase in wettability, resulting in a better water uptake and in an enhancement of growth parameters. The investigated DCSBD plasma source provides significant technical advantages and application potential for seed surface finishing without the use of hazardous chemicals.     

Some results on starlike trees and sunlike graphs

A tree is called starlike if it has exactly one vertex of degree greater than two. In [4] it was proved that two starlike treesG andH are cospectral if and only if they are isomorphic. We prove here that there exist no two non-isomorphic Laplacian cospectral starlike trees. Further, letG be a simple graph of ordern with vertex setV(G)={1,2, …,n} and letH={H ₁,H ₂, ...H _n } be a family of rooted graphs. According to [2], the rooted productG(H) is the graph obtained by identifying the root ofH _i with thei-th vertex ofG. In particular, ifH is the family of the paths $P_{k_1 } , P_{k_2 } , ..., P_{k_n } $ with the rooted vertices of degree one, in this paper the corresponding graphG(H) is called the sunlike graph and is denoted byG(k ₁,k ₂, …,k _n ). For any (x ₁,x ₂, …,x _n ) ∈I _* ⁿ , whereI _*={0,1}, letG(x ₁,x ₂, …,x _n ) be the subgraph ofG which is obtained by deleting the verticesi ₁, i₂, …,i _j ∈ V(G) (0≤j≤n), provided that $x_{i_1 } = x_{i_2 } = ... = x_{i_j } = 0$ . LetG(x ₁,x ₂,…, x _n] be the characteristic polynomial ofG(x ₁,x ₂,…, x _n ), understanding thatG[0, 0, …, 0] ≡ 1. We prove that $$G[k_1 , k_2 ,..., k_n ] = \Sigma _{x \in ^{I_ * ^n } } \left[ {\Pi _{i = 1}^n P_{k_i + x_i - 2} (\lambda )} \right]( - 1)^{n - (\mathop \Sigma \limits_{i = 1}^n x_i )} G[x_1 , x_2 , ..., x_n ]$$ where x=(x ₁,x ₂,…,x _n );G[k ₁,k ₂,…,k _n ] andP _n (γ) denote the characteristic polynomial ofG(k ₁,k ₂,…,k _n ) andP _n , respectively. Besides, ifG is a graph with λ₁(G)≥1 we show that λ₁(G)≤λ₁(G(k ₁,k ₂, ...,k _n )) < for all positive integersk ₁,k ₂,…,k _n , where λ₁ denotes the largest eigenvalue.     

On Integral Graphs Which Belong to the Class

Let G be a simple graph and let ¯G denote its complement. A graph G is said to be integral if its spectrum contains integral values. If is integral with >1 and a>b, where mG denotes the m-fold union of the graph G, we show that it belongs to the class of integral graphs where (i) (kn ² s ²+km nst+ms,km ² t ²+kmnst+ms)= such that (m,)=1 and (s,)=1; (ii) k,m,n,s,t,z such that (k,m)=1, (k,s)=1, (m,n)=1, (m,s)=1, (n,t)=1 and (s,t)=1; and (iii) ns>m t. Acknowledgments.The author is very grateful to the referees for their valuable comments and suggestions concerning this paper.1991 Mathematics Subject Classification. 05 C 50     

Generalized Farey Trees,Transfer Operators and Phase Transitions

We consider a family of Markov maps on the unit interval, interpolating between the tent map and the Farey map. The latter is not uniformly expanding. Each map being composed of two fractional linear transformations, the family generalizes many particular properties which for the case of the Farey map have been successfully exploited in number theory. We analyze the dynamics through the spectral properties of the generalized transfer operator. Application of the thermodynamic formalism to the family reveals first and second order phase transitions and unusual properties like positivity of the interaction function.     

Optical characterization of polycarbonate: Influence of additives on optical properties

In laser transmission welding of thermoplastics the optical properties of the joining parts determine the quality of the welding result. Especially, the scattering of laser radiation in the transparent welding part has an impact on weld seam properties. This scattering is caused by additives. For polycarbonate (PC) with different additives the transmittance, the reflectance and the collimated transmittance are measured with a UV‐VIS‐NIR spectrometer. From this data, the optical properties, such as scattering coefficient, absorption coefficient, and anisotropy factor are calculated. The calculations are made with the aid of the four‐flux model of radiation transport in the diffusive approximation. The results show that the additives have a significant influence on the scattering coefficient. For most additives under consideration the scattering is forward directed, which means that most of the radiation is transmitted into the absorbing welding part. However, the power density distribution of the transmitted radiation may differ significantly from PC without additives. So, the weld seam may also differ due to different additives. © 2010 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 48: 451–455, 2010