Objective To develop a method for the determination of ox-carbazepine (OXC) and 10,11-dihydro-10-hydroxy carbamazepine (MHD) in human plasma by solid phase extraction ( SPE) and high-performance liquid chromatography ( HPLC) .Methods OXC and MHD in human plasma was extracted by SPE tubes.The OXC and MHD were determined by HPLC.The chromatographic conditions were as following:the column was Shim-Pack C18 (250 mm ×4.6 mm,5μm ) , the mobile phase consisted of methanol and water (44:56 ) with a flow rate of 1.0 mL·min -1 , the injection volume was 40 μL, temperature was 25 ℃, detective wavelength was set 240 nm.Results OXC and MHD were in good linearity between 0.05-20 μg·mL-1 and 0.05-20 μg·mL-1 , respectively. Limit of detection for OXC and MHD were 25 , 50 ng·mL-1 ( S/N≥3 ) , respectively.Average recovery of extraction for OXC and MHD were 87.9%and 88.1%, respectively.Average method of extraction for OXC and MHD were 98.2%and 96.7%, respectively. Average between-day RSD for OXC and MHD were 6.5%and 5.9%as well as 4.8%and 4.0%of within-day RSD, respectively.Conclusion The established method for determination of OXC and MHD in human plasma by SPE is suitable for clinical application with high sensitivity, simple operation, high accuracy and reliability.

With prominent medicinal value, Gelsemium elegans has been overexploited, resulting in the reduction of the wild resource. As a result, artificial cultivation turns out to be a solution. However, this medicinal species is intolerant to low temperature, and thus genes responding to the low temperature are important for the cultivation of this species. Based on the transcriptome database of G. elegans at 4 ℃, 29 differentially expressed GeERF genes were identified. Bioinformatics analysis of 21 GeERF gene sequences with intact open reading frames showed that 12 and 9 of the GeERF proteins respectively clustered in DREB subgroup and ERF subgroup. GeDREB1 A-1-GeERF6 B-1, with molecular weight of 23.78-50.96 kDa and length of 212-459 aa, were all predicted to be hydrophilic and in nucleus. Furthermore, the full-length cDNA sequence of GeERF2B-1 was cloned from the leaves of G. elegans. Subcellular localization suggested that GeERF2B-1 was located in the nucleus. According to the quantitative reverse-transcription PCR(qRT-PCR), GeERF2B-1 showed constitutive expression in roots, stems, and leaves of G. elegans, and the expression was the highest in roots. In terms of the response to 4 ℃ treatment, the expression of GeERF2B-1 was significantly higher than that in the control and peaked at 12 h, suggesting a positive response to low temperature. This study lays a scientific basis for the functional study of GeERF transcription factors and provides gene resources for the improvement of stress resistance of G. elegans.
DNA, Complementary ; Gene Expression Regulation, Plant ; Phylogeny ; Plant Proteins/metabolism* ; Temperature ; Transcription Factors/metabolism*