[Preparation and chromatographic performance of cardanol-bonded silica stationary phase].

As green, less toxic, widely available, and site-rich functional ligands, natural products are widely used for the development of chromatographic stationary phases. In this work, a novel stationary phase, cardanol-bonded on silica (CBS) was prepared using γ-glycidoxypropyltrimethoxysilane (KH-560) as the coupling reagent and cardanol as the functional ligand. The synthesized stationary phase was characterized by Fourier transform-infrared spectra (FT-IR), thermogravimetric analysis (TGA), elemental analysis (EA), and N2 adsorption-desorption analysis. The results revealed that cardanol was successfully immobilized on the surface of spherical silica by the ring-opening reaction of the epoxy groups in phenolic hydroxyl. The retention mechanism and chromatographic performance of the CBS column were further evaluated and compared with those of a commercial C18 column using different classes of analytes, e. g., Tanaka standard test mixtures, alkylbenzenes, polycyclic aromatic hydrocarbons (PAHs), phenols, and aromatic positional isomers. The retention of alkylbenzenes under different chromatographic conditions revealed that the CBS column was a typical reversed-phase liquid chromatographic column, similar to the commercial C18 column. From the results of the Tanaka test, it was concluded that CBS could provide various interactions for different solutes e. g., hydrogen bonding and π-π interactions, along with hydrophobic interactions. The synergistic effects resulting from the aromatic rings, the hydroxyl and alkyl linkers in the new stationary phase improved the separation selectivity via multiple retention mechanisms. Based on these interactions, different solute probes such as hydrophobic alkylbenzenes, PAHs, and phenols were successfully separated in the reversed-phase liquid chromatography (RPLC) mode. For example, the aromatic positional isomers o-terthenyl, m-terphenyl, and triphenylene were used to investigate the chromatographic performance of the CBS column. These PHAs were baseline separated with good peak shapes. The resolution of m-terphenyl and triphenylene was as high as 6.81, while the two isomers could not be separated on the C18 column under the same chromatographic conditions. The repeatability and column stability of the CBS column was evaluated, and excellent repeatability and column stability were observed. The relative standard deviations (RSDs) of the retention time, peak area, and peak height for alkylbenzenes with 10 replicate injections were 0.052%-0.079%, 0.104%-0.847%, and 0.081%-0.272%, respectively. Traditional Chinese medicines have contributed notably to the Chinese civilization and human health. However, the complicated chemical compositions, unclear medicinal action mechanisms, and low purification efficiency for the traditional Chinese medicines have limited further development. Therefore it is necessary to establish an efficient, simple and feasible method for the separation and purification of herbal medicines. HPLC has been widely used in traditional Chinese medicines for the separation and detection of various components. In order to explore the CBS column for analysis of the traditional Chinese medicines, the ethanol extracts of fruits of Evodiae fructus and Camptotheca acuminata were used to test the separation performance of this column. The resolution of camptothecin from the preceding and following impurity peaks was 4.23 and 2.71. The resolution between evodiamine and rutaecarpin was 5.43, while the resolution from the adjacencies of impurity peaks was 2.20 and 1.69, respectively. The above mentioned results indicated that the CBS column shows good separation performance for the main active ingredients in the ethanolic extracts of these drugs, this validating its great potential for the analysis of real samples. Overall, the present study not only provides a new approach for the preparation of chromatographic stationary phases but also opens a new possibility for the separation and purification of camptothecin and evodiamine in real samples. This is an extension of the application of cardanol to chromatographic separation materials.

[The relationships among HBV genotypes, HBV DNA levels and histopathological features in the livers of familial grouped hepatitis B patients in the Tianjin area].

OBJECTIVE: To analyse the relationships among HBV genotypes, HBV DNA levels and histopathological features of the livers of familial grouped hepatitis B patients in the Tianjin area. METHODS: One hundred familial grouped hepatitis B (CHB) patients were enrolled in this study. Thirty-five of the 100 patients were chronic asymptomatic HBsAg carriers (ASC) and 65/100 were mild CHB patients. Their HBV genotypes and HBV-DNA levels were detected and liver biopsies were performed for analyzing the pathological features. RESULTS: Seven patients were of HBV genotype B (7%), and most of them had a HBV DNA level in the middle 10(3-5) copies/ml (57.14%). The histopathological features of the livers were of different degrees of injury. Eleven patients HBV was of genotype BC (11%); their HBV DNA levels were from 10(3-5) copies/ml (45.45%) to 10(6-7)copies/ml (36.36%). Their liver pathology showed slight or severe injuries (< or = G2 90.91%, < or = S(2) 81.82%). Eighty-two patients HBV was of genotype C (82%), and among the 82, 29 were ASC and 53 were CHB. Among the ASC, most of them had a high HBV DNA level (72.41%), and all of them had different degrees of liver injury. Among the CHB, their HBV DNA levels were 10(6-7) copies/ml (39.62%) and more than or equal 10(8) copies/ml (49.06%). The liver histopathological features were > or = G(2) in 38 patients (71.70%), and > or = S(2) in 25 patients (47.17%). CONCLUSIONS: (1) In the majority, HBV of the family gathered hepatitis B patients living in Tianjin is of genotype C and they have a high HBV-DNA level and severe liver pathological injuries. These features of the family gathered hepatitis B patients are the main factors causing the unfavorable prognosis of the patients. (2) There is inflammation of different degrees in the livers and high HBV DNA levels in all the family gathered ASC patients. Antiviral therapy should be planned according to the pathological features in patients livers. (3) Liver biopsies should be performed routinely before their antiviral therapy.

Chemiluminescence detection of isoniazid using Ru(phen)3(2+)-isoniazid-Ce(IV) system.

In our experiment, it was observed that isoniazid could enhance the chemiluminescence (CL) emission of tris-(1,10-phenanthroline)ruthenium(II) (Ru(phen)3(2+))-cerium(IV) (Ce(IV)) system and this enhancement effect was dependent on the concentration of isoniazid, based on which, a novel CL system was established for the detection of isoniazid. Under the optimum experimental conditions, the dynamic range and detection limit are 7.0 x 10(-2) to 6.5 microg ml(-1) and 2.5 x 10(-2) microg ml(-1), respectively. The R.S.D. is 3.4% (n = 11). The proposed method has been applied to detect the content of isoniazid in the injection solution with satisfactory results. The possible mechanism of the CL reaction was studied.