Cell Wall-bound p-Coumaric and Ferulic Acid Analysis
细胞壁结合的p-香豆和阿魏酸的分析
发布: 2016年10月05日第6卷第19期 DOI: 10.21769/BioProtoc.1955 浏览次数: 7754
评审: Marisa RosaAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Nov 2015
Abstract
Hydroxycinnamic acids, such as p-coumaric acid and ferulic acid, are a major class of compounds derived from the phenylpropanoid pathway. These compounds are widely conserved in plants and primarily accumulate in the secondary cell wall. They serve as important structural components that contribute to the overall strength and rigidity of plant cell walls and are also potent antioxidants valued for nutritional consumption. This protocol describes a method for analyzing hydroxycinnamic acids that are released after incubation under alkaline conditions.
Keywords: Cell wall (细胞壁)
P-coumaric acid (香豆酸)
Ferulic acid (阿魏酸)
Materials and Reagents
- 50 ml conical tubes (MIDSCI, catalog number: C50B )
- 2 ml microfuge tubes (MIDSCI, catalog number: AVX-T-20-C )
- Arabidopsis inflorescence tissue
- Liquid nitrogen
- Ethanol, pure (Sigma-Aldrich, catalog number: E7023 )
- Acetone (Sigma-Aldrich, catalog number: 320110 )
- 1 N NaOH (Sigma-Aldrich, catalog number: S8045 )
- 3,4,5-trimethoxycinnamic acid (Sigma-Aldrich, catalog number: T70408 )
- HCl (concentrated hydrochloric acid) (36-38%) (Sigma-Aldrich, catalog number: H1758 )
- Ethyl acetate (Sigma-Aldrich, catalog number: 270989 )
- Acetonitrile (Sigma-Aldrich, catalog number: 271004 )
- 0.1% formic acid (Sigma-Aldrich, catalog number: 94318 )
- p-coumaric acid (Sigma-Aldrich, catalog number: C9008 )
- Ferulic acid (Sigma-Aldrich, catalog number: 128708 )
- 50% methanol (Sigma-Aldrich, catalog number: 34860 )
Equipment
- Mortar and pestle
- Plastic spoon (appropriate for handling small volumes)
- Shaking incubator, 37 °C
- Centrifuge with a rotor that can accommodate 50 ml conical tubes and can reach 10,000 x g
- Microcentrifuge
- Vortex
- Drying oven, 70 °C
- Water bath, 65 °C
- Chemical scoop (appropriate for handling small volumes)
- Micropipette and tips (Mettler-Toledo, catalog number: L-STARTXLS+ )
- Repeating pipettor with adaptor syringe for dispensing 200 µl (Mettler-Toledo, model: AR-E1 )
- Speed Vac
- Analytical balance
- HPLC system with UV detector and autosampler (Shimadzu prominence modular HPLC: system controller CBM-20A, solvent delivery unit LC-20A, auto-sampler SIL-20A, column oven CTO-20A, UV-VIS detector SPD-20A)
- HPLC glass vials (VWR, catalog number: 89220-128 )
- Shim-pack XR-ODS column (column dimensions 3.0 x 75 mm, bead size 2.2 µm) (Shimadzu, model: Shim-pack XR-ODS )
Procedure
文章信息
版权信息
© 2016 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Anderson, N. (2016). Cell Wall-bound p-Coumaric and Ferulic Acid Analysis. Bio-protocol 6(19): e1955. DOI: 10.21769/BioProtoc.1955.
- Anderson, N. A., Bonawitz, N. D., Nyffeler, K. and Chapple, C. (2015). Loss of FERULATE 5-HYDROXYLASE leads to mediator-dependent inhibition of soluble phenylpropanoid biosynthesis in Arabidopsis. Plant Physiol 169(3): 1557-1567.
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分类
生物化学 > 其它化合物 > 酸
植物科学 > 植物生物化学 > 其它化合物
植物科学 > 植物细胞生物学 > 组织分析
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