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Studies of Preparation of L-borneol and Essential Oil from the Leaves of Blumea Balsamifera and Their Antioxidant and Antimicrobial Activities

Author: WangYuanHui
Tutor: WangHongXin
School: Jiangnan University
Course: Of Food Science
Keywords: Blumea balsamifera (L.) DC. L-borneol essential oil antioxidant activity antimicrobial activity
CLC: TS264.3
Type: PhD thesis
Year: 2013
Downloads: 111
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Abstract


Blumea balsamifera (L.) DC.(Asteraceae) is rich in terpenes and flavonoids and is afamous national spice and traditional Chinese medicine from Luodian County in China.Chinese Pharmacopoeia records that B. balsamifera is an important plant source of “Aipian”,natural (–)-borneol. The preparation of “Aipian” has last for over hundreds years in Guizhou,China.“Aipian” was prepared using the general hydrodistillation at a workshop in early period.With the continual progress of technology, the improvements (automation reconstruction)have been made in the production. However, there still are defects remaining in the process,such as low extraction rate, low yield, poor product quality, etc. Based on the previous workand physicochemical properties of L-borneol, a simple and high efficient preparation processwas designed to prepare high-purity L-borneol and obtain essential oil. The yields, chemicalcompositions, and functional activities of L-borneol and essential oil were studied. Results ofthis study are as follows:1. Relationship between raw material harvest period and active ingredientTo monitor the variation of material quality during harvest season and to arrangeproduction, the rapid determination of L-borneol in B. balsamifera leaves is indispensable.Six pretreatment methods were optimized to obtain the optimum extraction condition of eachmethod, including hydrodistillation-solvent extraction (HDSE), simultaneous distillationextraction (SDE), solvent extraction (SE), ultrasonic assisted extraction (UAE), microwaveassisted extraction (MAE), and ultrasonic-microwave assisted extraction (UMAE). Methodvalidation confirmed that the recovery of HDSE was the only one, which reached theaccuracy of simultaneous determination of L-borneol, camphor, and isoborneol in leaves, andthe values were between100%-105%. Furthermore, in the determination, the recovery ofL-borneol by UMAE was87%and met the criteria of determination. By comparison in time,energy, and solvent consumption, the consumption of UMAE was the lowest, microwavepower of100W, extraction time of30s, and UMAE was considered as the fastestpretreatment. Finally, fourteen batches of B. balsamifera leaves were determined byHDSE-GC and UMAE-GC, and the leaves harvested from October15to next January15hadhigher L-borneol content and were suitable as material.2. Isolation, purification, and identification of L-borneolBy comparison in the yield and efficiency of different methods, HD (Hydrodistillationwith vertical condensation tube) was considered to be the best extraction method, theoptimum conditions of HD were: the material and water ratio of1:50(g:mL), distillation timeof30min. The yield of L-borneol was0.85%(dry matter), and the recovery reached93%, andL-borneol content in crude extract was81%. L-borneol (Aipian) was separated from the crudeextract by sublimation at120°C for30min. The purity of L-borneol was91%, and the yieldwas0.83%(dry matter). The process was in free-solvent condition, and the recovery ofL-borneol reached91%. The recrystallization was used to purify L-borneol from “Aipian” andthe purity reached98%. Through repeatable recrystallization,82%of L-borneol in “Aipian”was recoveried, and the yield was0.75%(dry matter). The process is a simple and easy operation and cost less, so it is suitable for the preparation of high-purity L-borneol (98%).The optical activity and NMR showed that the purified L-borneol was really L-borneol.3. Extraction of essential oil of B. balsamifera leavesThe aroma components of B. balsamifera leaves were collected using headspace solidphase microextraction (HS-SPME). The yields of volatile oil through HDSE and SDE were1.00%and0.98%, respectively. The yield of hydrodistillation-extrusion (HDE) essential oilwas the lowest (only0.19%). The yield of two step hydrodistillation (TSHD) essential oil washigher than that of HDE essential oil, and reached0.32%. The aroma contained mainlysesquiterpenes, and among them caryophyllene content was the highest. Two volatiles oils andHDE essential oil had similar chemical composition and contained a large amout ofmonoterpenes. The differences between TSHD essential oil and other oils were verysignificant, and TSHD essential oil contained mainly sesquiterpenes.4. Antioxidant and antimicrobial activities of L-borneol and essential oilL-borneol did not exhibit the valuable antioxidative activity. By comparison, SDEvolatile oil had the best DPPH free radical scavenging capacity (IC50:10.07mg·mL-1). HDSEvolatile oil showed the higher inhibition capacity in BCB (IC50:2.45mg·mL-1). TSHDessential oil had the best performance in TBARS, and it was considered to be suitable asantioxidant in approximate organism environment. The purified L-borneol and standardcompound showed consistent antimicrobial activity, and the value of MIC was1mg·mL-1inantibacterial test, while it showed a better activity to Trichophyton rubrum (MIC:0.13mg·mL-1). HDSE volatile oil, SDE volatile oil, and HDE essential oil showed antimicrobialactivity against the tested bacteria and fungi, and the performance of antifungal activity wasbetter than antibacterial activity. TSHD essential oil had poor performance on the antibacterialactivity, but it had better performance in antifungal test. TSHD essential oil was suitable toinhibit the growth of fungi.5. Effect of L-borneol on cell wall and membraneEscherichia coli and Staphylococcus aureus were treated with L-borneol, and the growthcurves and time-kill curves of two bacterial were observed. The membrane potential of cellwas measured using DiBAC4(3). The integrity of cell membrane was observed usingpropidium iodide (PI). The result showed that L-borneol led the death of cell, but the integrityof membrane was not destroyed. L-borneol is in charge of the membrane potential of cell.When there is the existence of L-borneol, the cell depolarized, and membrane ion channel wasaffected, so that the normal growth of bacteria was destroyed to cause death.

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CLC: > Industrial Technology > Light industry,handicrafts > Food Industry > Brewing industry > Production of condiments > Spices, flavors
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