逆三相膜萃取生物碱分离纯化工艺研究

发布时间：2018-03-17 03:06

  本文选题：逆三相膜萃取　切入点：尼古丁　出处：《新疆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:建立逆三相膜萃取生物碱(尼古丁及氧化槐果碱),利用高效液相色谱法(HPLC)和紫外-可见分光光度法(UV)对提取物尼古丁和氧化槐果碱进行定性定量分析。方法:1.聚四氟乙烯膜(PTFE)作为萃取膜,有机溶剂为氯仿,水相和有机相体积比2.5∶1,0.01mol/L硫酸作为酸相萃取剂,1.25mol/L氢氧化钠作为碱相萃取剂,流速6mL/min,萃取时间25min。2.在单因素试验的基础上,采用L16(45)正交试验进一步考察条件,水相和有机相体积比为1∶1,0.3mol/L盐酸作为酸相萃取剂,0.75mol/L氢氧化钠作为碱相萃取剂,流速6mL/min、萃取时间60min。由直观分析可知,选取盐酸浓度(A)、氢氧化钠浓度(B)、提取时间(C)、体积比(D)4个因素,设计每个因素选取4个水平时,最佳提取工艺为A2B3C4D1,以盐酸浓度影响最大。结果:1.最佳实验条件下,尼古丁在25min内萃取率达到99.07%;高效液相色谱法定性定量分析尼古丁时色谱柱为C18柱(259mm×4.6mm),流动相为甲醇-0.2%磷酸水溶液(10∶90),进样量为10μL,柱温25℃,检测波长261nm,回归方程:Y=34014X+225.23,(r=0.9995,n=6),0.01~0.35μg/m L范围内线性关系良好,精密度RSD为0.98%,稳定性RSD为1.16%,重复性RSD为0.67%,平均加样回收率为99.26%。伊犁、巴楚、山东、库车和和田等五种产地1g烟草叶中尼古丁含量分别为24.43mg、23.12mg、21.39mg、13.72mg和5.93mg,伊犁烟草叶中尼古丁含量最高;2.最佳条件下氧化槐果碱在60min内萃取率达到98.21%。高效液相色谱法测定条件:Shim-packVP-ODS色谱柱(250mm×4.6mm,5μm),流动相为甲醇-0.2%磷酸水溶液(7∶93),梯度洗脱,流速1mL/min,进样量5μL,柱温30℃,检测波长221nm,在0.01~0.7mg/mL范围内线性关系良好,回归方程为Y=1610.90X-190.06,(r=0.9978,n=6),精密度RSD为0.93%,稳定性RSD为0.88%,重复性RSD为1.88%,平均回收率为97.35%。结论:1.通过逆三相膜循环萃取分离系统,萃取的尼古丁和氧化槐果碱纯度高、操作简单、有机溶剂消耗量低等特点,可用于其他生物碱、皂苷类和稀土金属离子的提取分离。
[Abstract]:Objective: to establish an inverse three-phase membrane extraction of alkaloids (nicotine and oxysophorine), and to determine the qualitative and quantitative analysis of nicotine and oxysophorine by high performance liquid chromatography (HPLC) and ultraviolet-visible spectrophotometry (UV). Polytetrafluoroethylene (PTFE) as an extraction membrane, The organic solvent is chloroform, the volume ratio of aqueous phase to organic phase is 2.5: 10.01mol / L sulfuric acid as acid phase extractant 1.25mol / L sodium hydroxide as alkali phase extractant, the flow rate is 6ml / min, the extraction time is 25min. 2. On the basis of single factor test, the conditions for further investigation are further investigated by L1645 orthogonal test. The volume ratio of hydrochloric acid to organic phase is 1: 1 / 1, 0.3 mol / L hydrochloric acid is used as acid phase extractant 0.75 mol / L sodium hydroxide as alkali phase extractant, the flow rate is 6 mL / min and the extraction time is 60 min. When each factor was selected at 4 levels, the optimum extraction process was A 2B 3C 4D 1, the concentration of hydrochloric acid was the most important. The extraction rate of nicotine reached 99.07% in 25 min, the chromatographic column for quantitative analysis of nicotine was C18 column (259mm 脳 4.6 mm), the mobile phase was methanol-0.2% phosphoric acid solution (10: 90m), the sample quantity was 10 渭 L, and the column temperature was 25 鈩，

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