【摘要】：本文主要考察了兩類選擇性試劑在高速逆流色譜(HSCCC)選擇性分離揮發(fā)油中化合物的應用,對甲基-β-環(huán)糊精與莪術油的包結作用以及色譜保留機制以及相關理論進行了探討。兩類選擇性試劑分別為環(huán)糊精類衍生物β-環(huán)糊精(β-CD)、甲基-β-環(huán)糊精(Me-β-CD)和羥丙基-β-環(huán)糊精(HP-β-CD),以及金屬離子硝酸銀。選擇了7種中藥揮發(fā)油作為選擇性分離的研究對象,包括莪術油、川芎揮發(fā)油、肉豆蔻揮發(fā)油、木香揮發(fā)油、當歸揮發(fā)油、青蒿揮發(fā)油、小茴香揮發(fā)油。首先,通過液液分配萃取實驗考察并確定了使用高速逆流色譜選擇性分離揮發(fā)油的兩相溶劑體系為正己烷-水相(1:1,v/v),其中水相中含有0.1mol/Lβ-CD/Me-β-CD/HP-β-CD。采用從尾到頭的洗脫方式用制備型高速逆流色譜分別分離了7種揮發(fā)油。將逆流色譜分離得到的收集液用制備型薄層色譜進行后處理,對最終樣品進行稱量并用氣質聯(lián)用色譜儀(GC-MS)和核磁共振氫譜(1H NMR)檢測。結果表明,逆流色譜選擇性分離得到了7種單體化合物:用Me-β-CD和HP-β-CD作為選擇性試劑從莪術油中分別分離得到了吉馬酮和郁金烯酮;用Me-β-CD和HP-β-CD作為選擇性試劑從川芎揮發(fā)油中都分離到了洋川芎內酯A;分別用β-CD和HP-β-CD作為選擇性試劑從肉豆蔻揮發(fā)油中分離得到了α-萜品醇和甲基丁香酚;用HP-β-CD作為選擇性試劑從木香揮發(fā)油中分離得到了反式紫羅蘭酮;用Me-β-CD作為選擇性試劑從當歸揮發(fā)油中分離得到了蒿苯內酯。其次,通過硅膠柱色譜用石油醚作為洗脫劑可以將β-欖香烯的百分含量從16.5%提高至46.1%。通過液液分配萃取實驗確定了使用高速逆流色譜選擇性分離莪術油中β-欖香烯的兩相溶劑體系為正己烷-甲醇-水(2:1.5:0.5,v/v/v),其中在下相中加入0.15mol L-1硝酸銀。采用從尾到頭的洗脫方式用制備型高速逆流色譜從莪術油中分離得到了純度大于99%的β-欖香烯。與普通的高速逆流色譜法相比大大縮短了分離時間,提高了樣品純度。最后,選擇莪術油與Me-β-CD作為研究對象,用高效液相色譜法分析了莪術油中倍半萜類化合物在碳十八色譜柱上的洗脫規(guī)律、分析物與Me-β-CD的包結常數(shù)以及其他相關熱力學常數(shù)。主要的分析物為吉馬酮、呋喃二烯和β-欖香烯。液相色譜條件為甲醇-水(90:10,v/v),由于流動相中甲醇比例較大,考慮了甲醇與Me-β-CD的包結作用。結果表明,三種倍半萜類化合物與Me-β-CD的包結比在濃度為0-9mmol L-1時均為1:1。用Van't Hoff方程計算了分析物與Me-β-CD以及分析物與固定相相互作用的焓變與熵變。通過在HSCCC中使用選擇性試劑,建立了選擇性分離揮發(fā)油的分離方法,該方法可大大提高HSCCC中目標成分色譜峰之間的分離度,大大縮短了分離時間,有效提高了分離純度。上述研究對逆流色譜選擇性分離揮發(fā)油中化合物具有指導性意義。
[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, 偽 -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.
【學位授予單位】：浙江工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：O652.6;R284

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