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论文题目: Quantification of plant cell wall monosaccharides by reversed-phase liquid chromatography with 2-aminobenzamide pre-column derivatization and a non-toxic reducing reagent 2-picoline borane
英文论文题目: Quantification of plant cell wall monosaccharides by reversed-phase liquid chromatography with 2-aminobenzamide pre-column derivatization and a non-toxic reducing reagent 2-picoline borane
第一作者: Fang, JJ; Qin, GC; Ma, J; She, YM
英文第一作者: Fang, JJ; Qin, GC; Ma, J; She, YM
联系作者: She, YM (reprint author), Chinese Acad Sci, Shanghai Ctr Plant Stress Biol, Shanghai 201602, Peoples R China.
英文联系作者: She, YM (reprint author), Chinese Acad Sci, Shanghai Ctr Plant Stress Biol, Shanghai 201602, Peoples R China.
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发表年度: 2015
卷: 1414
期:
页码: 122-128
摘要: In this report, we described a sensitive method for quantifying plant cell wall monosaccharides using chemical derivatization, reversed-phase high performance liquid chromatography separation with ultraviolet detection (HPLC-UV). Monosaccharides were derivatized with 2-aminobenzamide (2-AB) by reductive amination to increase the hydrophobicity and detected by ultraviolet absorption for HPLC-UV analysis. A non-toxic reductant, 2-picoline borane was utilized to replace the traditionally used sodium cyanoborohydride (NaCNBH3) to avoid the formation of toxic by-products. Experimental conditions were optimized using glucose as a model system to achieve a high reaction yield of 99%. Under the optimized conditions, we demonstrated that the derivatization yields of several saccharides with 2-AB using 2-picoline borane were all slightly higher than those observed using NaCNBH3. In plants, cell wall monosaccharides consist of arabinose, fucose, galactose, galacturonic acid, glucose, glucuronic acid, mannose, rhamnose, and xylose. Using our method, we successfully quantified these monosaccharides from Arabidopsis cell wall by HPLC-UV, and we obtained a good linearity at a wide dynamic range over five orders (1 pmol through 10 nmol of injection amount), a detection limit of similar to 0.1 pmole, and a high precision and accuracy. (C) 2015 Elsevier B.V. All rights reserved.
英文摘要: In this report, we described a sensitive method for quantifying plant cell wall monosaccharides using chemical derivatization, reversed-phase high performance liquid chromatography separation with ultraviolet detection (HPLC-UV). Monosaccharides were derivatized with 2-aminobenzamide (2-AB) by reductive amination to increase the hydrophobicity and detected by ultraviolet absorption for HPLC-UV analysis. A non-toxic reductant, 2-picoline borane was utilized to replace the traditionally used sodium cyanoborohydride (NaCNBH3) to avoid the formation of toxic by-products. Experimental conditions were optimized using glucose as a model system to achieve a high reaction yield of 99%. Under the optimized conditions, we demonstrated that the derivatization yields of several saccharides with 2-AB using 2-picoline borane were all slightly higher than those observed using NaCNBH3. In plants, cell wall monosaccharides consist of arabinose, fucose, galactose, galacturonic acid, glucose, glucuronic acid, mannose, rhamnose, and xylose. Using our method, we successfully quantified these monosaccharides from Arabidopsis cell wall by HPLC-UV, and we obtained a good linearity at a wide dynamic range over five orders (1 pmol through 10 nmol of injection amount), a detection limit of similar to 0.1 pmole, and a high precision and accuracy. (C) 2015 Elsevier B.V. All rights reserved.
刊物名称: JOURNAL OF CHROMATOGRAPHY A
英文刊物名称: JOURNAL OF CHROMATOGRAPHY A
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学科: Biochemistry & Molecular Biology; Chemistry
英文学科: Biochemistry & Molecular Biology; Chemistry
影响因子: 4.169
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论文类别: Article
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