基于络合作用的逆流色谱分离挥发油化学成分及相关理论研究
发布时间：2018-08-07 07:51
【摘要】：本文主要考察了两类选择性试剂在高速逆流色谱(HSCCC)选择性分离挥发油中化合物的应用,对甲基-β-环糊精与莪术油的包结作用以及色谱保留机制以及相关理论进行了探讨。两类选择性试剂分别为环糊精类衍生物β-环糊精(β-CD)、甲基-β-环糊精(Me-β-CD)和羟丙基-β-环糊精(HP-β-CD),以及金属离子硝酸银。选择了7种中药挥发油作为选择性分离的研究对象,包括莪术油、川芎挥发油、肉豆蔻挥发油、木香挥发油、当归挥发油、青蒿挥发油、小茴香挥发油。首先,通过液液分配萃取实验考察并确定了使用高速逆流色谱选择性分离挥发油的两相溶剂体系为正己烷-水相(1:1,v/v),其中水相中含有0.1mol/Lβ-CD/Me-β-CD/HP-β-CD。采用从尾到头的洗脱方式用制备型高速逆流色谱分别分离了7种挥发油。将逆流色谱分离得到的收集液用制备型薄层色谱进行后处理,对最终样品进行称量并用气质联用色谱仪(GC-MS)和核磁共振氢谱(1H NMR)检测。结果表明,逆流色谱选择性分离得到了7种单体化合物:用Me-β-CD和HP-β-CD作为选择性试剂从莪术油中分别分离得到了吉马酮和郁金烯酮;用Me-β-CD和HP-β-CD作为选择性试剂从川芎挥发油中都分离到了洋川芎内酯A;分别用β-CD和HP-β-CD作为选择性试剂从肉豆蔻挥发油中分离得到了α-萜品醇和甲基丁香酚;用HP-β-CD作为选择性试剂从木香挥发油中分离得到了反式紫罗兰酮;用Me-β-CD作为选择性试剂从当归挥发油中分离得到了蒿苯内酯。其次,通过硅胶柱色谱用石油醚作为洗脱剂可以将β-榄香烯的百分含量从16.5%提高至46.1%。通过液液分配萃取实验确定了使用高速逆流色谱选择性分离莪术油中β-榄香烯的两相溶剂体系为正己烷-甲醇-水(2:1.5:0.5,v/v/v),其中在下相中加入0.15mol L-1硝酸银。采用从尾到头的洗脱方式用制备型高速逆流色谱从莪术油中分离得到了纯度大于99%的β-榄香烯。与普通的高速逆流色谱法相比大大缩短了分离时间,提高了样品纯度。最后,选择莪术油与Me-β-CD作为研究对象,用高效液相色谱法分析了莪术油中倍半萜类化合物在碳十八色谱柱上的洗脱规律、分析物与Me-β-CD的包结常数以及其他相关热力学常数。主要的分析物为吉马酮、呋喃二烯和β-榄香烯。液相色谱条件为甲醇-水(90:10,v/v),由于流动相中甲醇比例较大,考虑了甲醇与Me-β-CD的包结作用。结果表明,三种倍半萜类化合物与Me-β-CD的包结比在浓度为0-9mmol L-1时均为1:1。用Van't Hoff方程计算了分析物与Me-β-CD以及分析物与固定相相互作用的焓变与熵变。通过在HSCCC中使用选择性试剂,建立了选择性分离挥发油的分离方法,该方法可大大提高HSCCC中目标成分色谱峰之间的分离度,大大缩短了分离时间,有效提高了分离纯度。上述研究对逆流色谱选择性分离挥发油中化合物具有指导性意义。
[Abstract]:The application of two kinds of selective reagents in the selective separation of volatile oil by high speed countercurrent chromatography (HSCCC) was studied. The inclusion of methyl 尾 -cyclodextrin with zedoary turmeric oil, the retention mechanism and the related theories were discussed. The two kinds of selective reagents are cyclodextrin derivatives 尾 -cyclodextrin (尾 -CD), methyl- 尾 -cyclodextrin (Me- 尾 -CD) and hydroxypropyl- 尾 -cyclodextrin (HP- 尾 -CD), as well as metal ion silver nitrate. Seven kinds of essential oils of traditional Chinese medicine were selected for selective separation, including zedoary turmeric oil, Chuanxiong volatile oil, nutmeg essential oil, wood volatile oil, Angelica sinensis volatile oil, Artemisia annua volatile oil and fennel volatile oil. Firstly, the two-phase solvent system for selective separation of volatile oil by high speed countercurrent chromatography was investigated by liquid-liquid partition extraction experiment. The two-phase solvent system was n-hexane / water phase (1: 1 v / v), in which the aqueous phase contained 0.1mol/L 尾 -CD-CDR / Me- 尾 -CD-HP- 尾 -CD. Seven kinds of volatile oils were separated by high speed countercurrent chromatography from tail to head. The collected liquid separated by countercurrent chromatography was treated by thin-layer chromatography. The final sample was weighed and detected by GC-MS and 1H NMR. The results showed that seven monomers were isolated from zedoary turmeric oil selectively by countercurrent chromatography. Gimarone and tugolenone were separated from zedoary turmeric oil by Me- 尾 -CD and HP- 尾 -CD as selective reagents. Ligustrade A was isolated from essential oil of Ligusticum chuanxiong with Me- 尾 -CD and HP- 尾 -CD as selective reagents, 伪 -terpenol and methyleugenol were isolated from essential oil of nutmeg with 尾 -CD and HP- 尾 -CD as selective reagents, respectively. Using HP- 尾 -CD as selective reagent, trans-ionone was isolated from volatile oil of Radix angelicae Sinensis, and artemiolactone was separated from volatile oil of Angelica sinensis with Me- 尾 -CD as selective reagent. Secondly, the content of 尾 -elemene was increased from 16.5% to 46.1% by using petroleum ether as eluent in silica gel column chromatography. The two-phase solvent system of selective separation of 尾 -elemene in zedoary turmeric oil by high speed countercurrent chromatography was determined by liquid-liquid partition extraction experiment as n-hexane-methanol-water (2: 1.5: 0.5 v / v), in which 0.15mol L-1 silver nitrate was added to the lower phase. 尾 -elemene with purity more than 99% was isolated from Zedoary turmeric oil by high speed countercurrent chromatography with tail to head elution. Compared with conventional high-speed counter-current chromatography, the separation time is greatly shortened and the purity of the sample is improved. Finally, the elution of sesquiterpenoids in zedoary turmeric oil and Me- 尾 -CD on C _ (18) chromatographic column, the inclusion constants and other thermodynamic constants of sesquiterpenes in zedoary oil were analyzed by HPLC. The main analytes are gemanone, furandiene and 尾-elemene. The liquid chromatographic condition is methanol-water (90: 10 v / v). Because the ratio of methanol in mobile phase is large, the inclusion of methanol with Me- 尾 -CD is considered. The results showed that the inclusion ratio of three sesquiterpenes to Me- 尾 -CD was 1: 1 at the concentration of 0-9mmol L-1. The enthalpy change and entropy change of the interaction between analyte and Me- 尾 -CD and between analyte and stationary phase are calculated by Van't Hoff equation. By using selective reagent in HSCCC, a selective separation method for volatile oil was established. The method can greatly improve the separation degree between the chromatographic peaks of the target components in HSCCC, shorten the separation time and improve the separation purity. These studies are of guiding significance for selective separation of compounds in volatile oil by countercurrent chromatography.
【学位授予单位】：浙江工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O652.6;R284
本文编号：2169359
[Abstract]:The application of two kinds of selective reagents in the selective separation of volatile oil by high speed countercurrent chromatography (HSCCC) was studied. The inclusion of methyl 尾 -cyclodextrin with zedoary turmeric oil, the retention mechanism and the related theories were discussed. The two kinds of selective reagents are cyclodextrin derivatives 尾 -cyclodextrin (尾 -CD), methyl- 尾 -cyclodextrin (Me- 尾 -CD) and hydroxypropyl- 尾 -cyclodextrin (HP- 尾 -CD), as well as metal ion silver nitrate. Seven kinds of essential oils of traditional Chinese medicine were selected for selective separation, including zedoary turmeric oil, Chuanxiong volatile oil, nutmeg essential oil, wood volatile oil, Angelica sinensis volatile oil, Artemisia annua volatile oil and fennel volatile oil. Firstly, the two-phase solvent system for selective separation of volatile oil by high speed countercurrent chromatography was investigated by liquid-liquid partition extraction experiment. The two-phase solvent system was n-hexane / water phase (1: 1 v / v), in which the aqueous phase contained 0.1mol/L 尾 -CD-CDR / Me- 尾 -CD-HP- 尾 -CD. Seven kinds of volatile oils were separated by high speed countercurrent chromatography from tail to head. The collected liquid separated by countercurrent chromatography was treated by thin-layer chromatography. The final sample was weighed and detected by GC-MS and 1H NMR. The results showed that seven monomers were isolated from zedoary turmeric oil selectively by countercurrent chromatography. Gimarone and tugolenone were separated from zedoary turmeric oil by Me- 尾 -CD and HP- 尾 -CD as selective reagents. Ligustrade A was isolated from essential oil of Ligusticum chuanxiong with Me- 尾 -CD and HP- 尾 -CD as selective reagents, 伪 -terpenol and methyleugenol were isolated from essential oil of nutmeg with 尾 -CD and HP- 尾 -CD as selective reagents, respectively. Using HP- 尾 -CD as selective reagent, trans-ionone was isolated from volatile oil of Radix angelicae Sinensis, and artemiolactone was separated from volatile oil of Angelica sinensis with Me- 尾 -CD as selective reagent. Secondly, the content of 尾 -elemene was increased from 16.5% to 46.1% by using petroleum ether as eluent in silica gel column chromatography. The two-phase solvent system of selective separation of 尾 -elemene in zedoary turmeric oil by high speed countercurrent chromatography was determined by liquid-liquid partition extraction experiment as n-hexane-methanol-water (2: 1.5: 0.5 v / v), in which 0.15mol L-1 silver nitrate was added to the lower phase. 尾 -elemene with purity more than 99% was isolated from Zedoary turmeric oil by high speed countercurrent chromatography with tail to head elution. Compared with conventional high-speed counter-current chromatography, the separation time is greatly shortened and the purity of the sample is improved. Finally, the elution of sesquiterpenoids in zedoary turmeric oil and Me- 尾 -CD on C _ (18) chromatographic column, the inclusion constants and other thermodynamic constants of sesquiterpenes in zedoary oil were analyzed by HPLC. The main analytes are gemanone, furandiene and 尾-elemene. The liquid chromatographic condition is methanol-water (90: 10 v / v). Because the ratio of methanol in mobile phase is large, the inclusion of methanol with Me- 尾 -CD is considered. The results showed that the inclusion ratio of three sesquiterpenes to Me- 尾 -CD was 1: 1 at the concentration of 0-9mmol L-1. The enthalpy change and entropy change of the interaction between analyte and Me- 尾 -CD and between analyte and stationary phase are calculated by Van't Hoff equation. By using selective reagent in HSCCC, a selective separation method for volatile oil was established. The method can greatly improve the separation degree between the chromatographic peaks of the target components in HSCCC, shorten the separation time and improve the separation purity. These studies are of guiding significance for selective separation of compounds in volatile oil by countercurrent chromatography.
【学位授予单位】：浙江工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O652.6;R284
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1 鲁梦霞;基于络合作用的逆流色谱分离挥发油化学成分及相关理论研究[D];浙江工业大学;2017年
2 韩纪良;三七姜挥发油对人鼻咽癌CNE-2细胞的侵袭和迁移抑制作用及其机制初步探讨[D];桂林医学院;2017年
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