Isolation of Triterpenes from Propolis (Bee Glue)
从蜂胶中分离三萜
发布: 2015年10月20日第5卷第20期 DOI: 10.21769/BioProtoc.1630 浏览次数: 14110
评审: Arsalan DaudiCindy AstAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Oct 2014
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Abstract
Propolis (bee glue) is a natural substance produced by bees upon collection of mainly plant resins. Bees use it as antiseptic sealing agent between honeycombs and to preserve the hive from external contamination. Numerous scientific studies have been published on the biological properties of propolis including its anti-inflammatory, anti-oxidant, immunostimulant, antitumour and antimicrobial activity. Different propolis chemotypes have been characterised based on the nature of the plant-derived substances present and the geographical origin of collection. Here, we describe the isolation of nine triterpenes from a sample of propolis originating from North-Western Cameroon. All compounds were identified following analysis of their spectroscopic data and comparison with previously published reports.
Materials and Reagents
- TLC silica gel 60 F254 plates (VWR International, catalog number: 1.05554.0001 )
- Wilmad® NMR tubes 5 mm diam., precision (Sigma-Aldrich, catalog number: Z274275 )
- Snap-cap vials (VWR International, catalog number: 548-0555 )
- Raw propolis (Nature’s Laboratory Ltd, catalog number: P5 )
- Ethanol 96% v/v (extra pure, Specified Laboratory Reagent) (Thermo Fisher Scientific, catalog number: 10162252 )
- Hexanes (for HPLC, 95% n-hexane approx) (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10703611 )
- Ethyl acetate, extra pure, Specified Laboratory Reagent (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10080130 )
- Sodium sulfate anhydrous, extra pure (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10606082 )
- Silica gel 60 (0.063–0.200 mm) (VWR International, catalog number: 1.07734.1000 )
- Silica gel 60H (VWR International, catalog number: 1.07736.1000 )
- 98% p-Anisaldehyde (Sigma-Aldrich, catalog number: A88107 )
- Acide sulfurique fumant (pure) (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10222282 )
- Acetic acid glacial (pure) (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10375020 )
- Methanol (extra pure, Specified Laboratory Reagent) (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10214490 )
- Sephadex® LH-20 (Sigma-Aldrich, catalog number: LH20100 )
- α-Amyrin (Sigma-Aldrich, catalog number: 53017 )
- Cycloartenol (Sigma-Aldrich, catalog number: 08172 )
- Lupeol (Sigma-Aldrich, catalog number: S957712 )
- Dichloromethane, extra pure, stabilised with amylene, Specified Laboratory Reagent (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10458210 )
- Chloroform-d (CDCl3) (Sigma-Aldrich, catalog number: 10080130 )
- Anisaldehyde-sulphuric acid reagent (see Recipes)
Equipment
- Heated ultrasonic water bath (340 W) (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10215332 )
- Büchi rotary evaporator, Rotavapor® R-210 (VWR International, catalog number: 531-0850 )
- UV viewing cabinet CC10 (Thermo Fisher Scientific, Fisher Scientific, catalog number: 11778201 )
- UV lamp UVG-11 230 V 50/60 Hz 4 W hand held short wave (Thermo Fisher Scientific, Fisher Scientific, catalog number: 11718241 )
- UV lamp UVL-21 230 V 50/60 Hz 4 W hand held long wave (Thermo Fisher Scientific, Fisher Scientific, catalog number: 11728241 )
- Analytical balance (VWR International, catalog number: 611-2267 )
- Precision balance (VWR International, catalog number: 611-2695 )
- Vacuum water jet pump (VWR International, catalog number: 181-9100 )
- Pear-shaped (separating) funnel (1,000 ml) (Scientific Glass Laboratories, catalog number: SFP1L )
- Round bottom flasks, 250 ml (Total number of 12) (Scientific Glass Laboratories, catalog number: FRS250/B19 or FRS250/B24 )
- Glass foot measuring cylinder with hexagonal base, borosilicate glass, class “B” (25 ml and 250 ml) (Scientific Glass Laboratories, catalog number: MCB/25 and MCB/250 )
- Conical Erlenmeyer flasks (500 ml) (Scientific Glass Laboratories, catalog number: FC500/B24 )
- Filter funnels, borosilicate (Scientific Glass Laboratories, catalog number: BFF75 and BFF100 )
- Rubber vacuum tubing NW8 (VWR international, catalog number: 189.3111 )
- Glass Pasteur pipettes 150 mm (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10006021 )
- Glass Pasteur pipettes 230 mm (Thermo Fisher Scientific, Fisher Scientific, catalog number: 10209381 )
- Glass micropipettes (homemade from glass Pasteur pipettes 230 mm using a Bunsen burner)
- Büchner filter with cone and thread, GL14, for vacuum liquid chromatography (VLC) (Scientific Glass Laboratories, catalog number: TBF/4 / B24 / POR3 )
- Twin trough chambers for TLC plates (VWR International, catalog number: 552-0011 )
- Glass atomiser reagent sprayer (VWR International, catalog number: 552-0031 )
- Glass chromatography columns (CC) with sintered discs (Scientific Glass Laboratories, catalog number: R2/40 )
- Duratool DO1600 hot air heat gun (Amazon)
- Whatman qualitative filter paper, grade 1 (Sigma-Aldrich, catalog number: WHA1001125 )
- Exactive Orbitrap mass spectrometer (MS) operating in a positive and negative electrospray ionisation (ESI) switching mode (Thermo Fisher Scientific)
- JEOL 505HA high resolution electron impact (HREI) mass spectrometer (MS) using direct probe at elevated temperature (110~160 ºC) at 70 eV (JEOL USA)
- JEOL Lambda Delta 400 NMR spectrometer (JEOL USA, model: JEOL Nuclear Magnetic Resonance Spectrometers )
Software
- Xcalibur software (version 2.2) for MS data processing (Thermo Fisher Scientific, model: Xcalibur software)
- Mestre Nova (MNova) software (version 8.0.0) for NMR processing (Mestrelab Research SL, model: version 8.0.0)
Procedure
文章信息
版权信息
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Kardar, M. N. and Seidel, V. (2015). Isolation of Triterpenes from Propolis (Bee Glue). Bio-protocol 5(20): e1630. DOI: 10.21769/BioProtoc.1630.
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分类
植物科学 > 植物生物化学 > 其它化合物
生物化学 > 其它化合物 > 萜类 > 三萜
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