【作者】 张勇；

【导师】 李章万；

【作者基本信息】 四川大学 ， 药物分析， 2005， 硕士

【摘要】 摘要一 哌嗪异黄酮的结构确证及质量标准研究 哌嗪异黄酮是一种全新合成的可望用于治疗骨质疏松的植物雌激素类药物,它是由异黄酮和哌嗪通过乙氧键连接而成。 本文采用多种波谱技术,包括:UV、IR、MS、NMR(¹H-NMR、¹³C-NMR、DEPT-¹³C-NMR、¹H-¹HCOSY、HMQC、HMBC)等,并结合元素分析对哌嗪异黄酮的结构进行了研究。归属了本化合物红外光谱中的主要特征吸收峰和质谱中的主要特征碎片离子。特别是利用各种2D-NMR技术,包括:氢-氢相关谱、碳-氢相关谱等确定了氢谱及碳谱的谱线归属。综合应用以上几种方法确证了哌嗪异黄酮的结构。 本文对哌嗪异黄酮原料的质量标准进行了研究,主要内容如下:（1） 采用Diamosil C₁₈（150mm×4.6mm i.d.,5μm）为分析柱;甲醇—0.025mol/L磷酸二氢钠和0.04mol/L十二烷基磺酸钠的混合溶液（80:20）（磷酸调节pH4.8）为流动相;于248.6nm处检测。建立了哌嗪异黄酮原料有关物质检查和含量测定的离子对液相色谱法。所建立的有关物质检查方法在2.08～8.32ug/ml范围内线性关系良好（r=0.9999）;最低检出量为0.2ng（S/N=3）。所建立的含量测定方法在19.7～78.8ug/ml范围内线性关系良好（r=0.9999）;日内和日间精密度分别为0.30%和0.96%。（2） 采用无水冰醋酸为溶剂,0.1mol/L高氯酸滴定液滴定,电位法指示终点,建立了哌嗪异黄酮原料含量测定的非水溶液滴定法;此法日内及日间精密度分别为0.34%和0.32%。（3） 采用硅胶GF₂₅₄为吸附剂;甲苯-甲醇（4:1）为展开剂;氨饱和;更多还原

【Abstract】 Piperazinylethylisoflavones ether is a new synthesized compound that can be used for the prevention and treatment of osteoporosis and other disease caused by osteoporosis.For further study on this substance, in this article, the structure of Piperazinylethylisoflavones ether was analyzed and the main characteristic adsorption bands in its IR spectral and fragment ions in its mass spectral are also assigned by using a series of spectral techniques including UV, IR, MS, NMR and the element analysis. Especially, several kinds of 2D-NMR techniques (e. g. 1H-NMR、 13C-NMR、 DEPT-13C-NMR、 1H-1HCOSY、 HMQC、 HMBC), which can provide valuable information, are used for confirmation of the assignment of its highly overlapping 1H-NMR and 13C-NMR signals. The combination of these techniques is applicable for the structural establishment of Piperazinylethylisoflavones ether.In addition, in this article, the quality standard of Piperazinylethylisoflavones ether was also studied: (1)ODS column as stationary phase, methanol and a mixture of 0. 025mol/L sodium phosphate monobasic and 0. 04mol/L sodium dedecyl sulfate(80:20) (adjusted with phosphoric acid to a pH 4.8) as mobile phase , 248. 6nm as detective wavelength to establish ion-pair HPLC method for the content determination and limit test of the relative substance. This established method of the relative substance limit test had good specification, linearity and precision within the range of 2. 08~8. 32ug/ml (r=0. 9999), the limit of detection is 0.2ng(S/N=3).The established content method had good linearity within the range of 19.7 ~ 78. 8ug/ml(r=0. 9999). The RSD with-in day and between-day were 0.30% and 0.96%, respectively. (2)Anhydrous acetic acid as solvent, 0.1mol/L perchloric acid as titrant, potentio-metric method indicating end-piont to establish non-aqueous titration method for determination of Piperazinylethylisoflavones ether. The established method is simple, accurate and precise. The RSD with-in day and between-day were 0.34% and 0.32%, respectively. (3)Silica GF254 as adsorptive reagent ; toluene-methanol (4:1) as the developer; chamber saturated with the developer and ammonia to establish the TLC method for the relative substance of Piperazinylethylisoflavones ether. (4) For the limit test of residual solvent in Piperazinylethylisoflavones ether drug substance, A capillary GC method was established using HP-1 (30m×0.53mm×0.33 μm) as the analytical column; 3mX0. 53mm fused silica capillary column as the guard column; nitrogen as the carrier gas with a flow rate of 20ml/min; the flow rate of air and hydrogen was 250ml/min and 35ml/min; the oven temperature held at 100℃; The injector anddetector temperatures were maintained at 220°C and 250°C; The split ratio was 1:1.The method had a good specification and sensitivity.(二) STUDY ON QUALITY STANDARD OF ISOFLURANE INJECTION At present, isoflurane is used widely as a sort of volatile general anesthetics. In the traditional way, isoflurane was presented into respiratory tract by respiration machine . With the development of general anesthetics, isoflurane can be injected directly into vein. In this article,we established a method for determination of isoflurane and its related substances in isoflurane injection. The isoflurane injection was dissolved with n-butanol, and then detected by capillary gas chromatograph. N-hexane was used as the internal standard. The linear range of determination of related substances was 60~ 180ug/ml.The regression equation was Y=0.0086X+0.0018 (r=0. 9996). The limit of detection was 60ng. The linear range of determination of isoflurane injection was 6 ~ 18mg/ml. The regression equation was Y=0.1035X-0.0023 ( r=0.9999). The average recoveries of the method was 99. 49%, RSD=1.1%. The RSD of the inter-day and intra-day variation were 0. 48% and 0. 87%, respectively. The method was simple, selective and reproducible for the assay of isoflurane in isoflurane injection.更多还原