高效液相色谱法测定玉米幼苗叶片中4种酚酸类化合物
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摘要
为研究干旱胁迫前后玉米幼苗叶片中4种酚类物质的变化,采用高效液相色谱法(HPLC),以Thermo Scientific Syncronis C18 (4.6 mm×250 mm,5μm)为分析柱,乙腈-0.5%乙酸溶液梯度洗脱,流速1.0 mL/min,检测波长280 nm,柱温30℃,对不同品种玉米幼苗叶片中的没食子酸、香豆酸、绿原酸、咖啡酸含量进行测定。结果表明,4种酚酸化合物的浓度在1～100 mg/L范围内与峰面积线性关系良好,相关系数R2均大于0.999,加标回收率和精密度RSD结果均满足分析要求;玉米幼苗叶片中香豆酸含量较高、没食子酸含量较少,干旱胁迫后除咖啡酸外其他3种酚酸含量均有下降;不同品种玉米幼苗叶片中4种酚类物质的含量不同,且干旱胁迫后酚酸的含量变化不同,与其对干旱的敏感程度有关。
The aim is to study the changes of four phenolic acids in the leaves of maize seedlings before and after drought stress. The content of gallic acid, coumalic acid, chlorogenic acid and caffeic acid were quantitatively determined by High Performance Liquid Chromatography(HPLC), Thermo Scientific Syncronis C18 column(4.6 mm×250 mm, 5 μm) as analytical column, acetonitrile-0.5% acetic acid solution with gradient elution, 1.0 mL/min flow rate, 280 nm detection wavelength and 30℃ column temperature. The results showed that the linear relationship between peak area and 1-100 mg/L gallic acid, coumalic acid, chlorogenic acid and caffeic acid was good, the correlation coefficients(R2) were greater than 0.999. Adding standard recovery and precision RSD fulfilled the analysis requirements. The coumalic acid content was relatively high and gallic acid content was relatively low. After drought stress, the contents of the other three phenolic acids decreased except caffeic acid. The contents of four phenolic compounds in different varieties of maize seedlings was different and changed differently under drought stress, which was related to its sensitivity to drought.
The aim is to study the changes of four phenolic acids in the leaves of maize seedlings before and after drought stress. The content of gallic acid, coumalic acid, chlorogenic acid and caffeic acid were quantitatively determined by High Performance Liquid Chromatography(HPLC), Thermo Scientific Syncronis C18 column(4.6 mm×250 mm, 5 μm) as analytical column, acetonitrile-0.5% acetic acid solution with gradient elution, 1.0 mL/min flow rate, 280 nm detection wavelength and 30℃ column temperature. The results showed that the linear relationship between peak area and 1-100 mg/L gallic acid, coumalic acid, chlorogenic acid and caffeic acid was good, the correlation coefficients(R2) were greater than 0.999. Adding standard recovery and precision RSD fulfilled the analysis requirements. The coumalic acid content was relatively high and gallic acid content was relatively low. After drought stress, the contents of the other three phenolic acids decreased except caffeic acid. The contents of four phenolic compounds in different varieties of maize seedlings was different and changed differently under drought stress, which was related to its sensitivity to drought.
引文
[1]王德信,杨晓莹.玉米幼苗对干旱胁迫的生理响应[J].贵州农业科学,2018,46(4):26-29.
[2]张凤启,赵霞,丁勇,等.玉米耐旱性研究进展[J].中国农学通报,2015,31(30):38-42.
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[7]白英俊,李国瑞,黄凤兰,等.活性氧与植物抗氧化系统研究进展[J].安徽农业科学,2017,45(36):1-3.
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[10] Anjum S A, Umair A, Mohsin T, et al. Drought Induced Changes in Growth, Osmolyte Accumulation and Antioxidant Metabolism of Three Maize Hybrids[J]. Frontiers in Plant Science,2017,8:69.
[11] Sarker U, Oba S. Drought stress enhances nutritional and bioactive compounds, phenolic acids and antioxidant capacity of Amaranthus leafy vegetable[J]. BMC Plant Biology,2018,18(1):258.
[12] Sun C X, Li M Q, Gao X X, et al. Metabolic response of maize plants to multifactorrial abiotic stresses[J]. Plant Biology,2016,18(suppl.1):120-129.
[13]成锴,苏晓慧,栗建枝,等.PEG-6000胁迫下玉米品种萌发期抗旱性鉴定与评价[J].玉米科学,2017,25(5):85-90.
[14]刘海芳,魏东伟,刘全军,等.太行菊不同器官中绿原酸和4种黄酮类物质含量研究[J].天然产物研究与开发,2013,25(5):646-651;671.
[15]谢昕.玉米苞叶多酚提取工艺优化[J].农产品加工,2018(7):47-51.
[16]苏贻娟,冯远娇,罗赐君,等.高效液相色谱法测定玉米叶片中的酚酸类化合物[J].玉米科学,2009,17(5):166-168.
[17]赵霞,刘诗慧,张国方,等.16个玉米杂交种的抗旱性评价[J].河南农业科学,2017,46(12):24-28.
[18]王娟,李德全,谷令坤.不同抗旱性玉米幼苗根系抗氧化系统对水分胁迫的反应[J].西北植物学报,2002(2):77-82.
[19]曾智,陈根洪,滕树锐,等.干旱胁迫下厚朴幼苗的生理指标及次生代谢产物对保水剂的响应[J].福建农林大学学报:自然科学版,2016,45(5):544-550.
[20]赵贝贝,叶蕴灵,王莉,等.银杏类黄酮响应非生物胁迫研究进展[J].扬州大学学报:农业与生命科学版,2018,39(3):106-112.
[21]马惠,王琦,赵鸣,等.非生物胁迫对棉花次生代谢及棉蚜种群消长的影响[J].棉花学报,2016,28(4):324-330.
[22] Hodaei M, Rahimmalek M, Arzani A, et al. The effect of water stress on phytochemical accumulation, bioactivecompounds and expression of key genes involved in flavonoid biosynthesisin Chrysanthemum morifolium L.[J]. Industrial Crops&Products,2018,120:295-304.
[23]杨婷.高效液相色谱法在中药鉴定中的应用分析及前景[J].中国化工贸易,2018,10(32):141.
[24]李颖畅,张明辉,马春颖,等.响应面法优化超声波辅助提取蓝莓叶多酚[J].食品工业科技,2015,36(15):234-238.
[2]张凤启,赵霞,丁勇,等.玉米耐旱性研究进展[J].中国农学通报,2015,31(30):38-42.
[3] Freire De Sousa D P, Braga B B, Gondim F A, et al. Increased drought tolerance in maize plants induced by H2O2is closely related to an enhanced enzymatic antioxidant system and higher soluble protein and organic solutes contents[J]. Theoretical and Experimental Plant Physiology,2016,28(3):297-306.
[4]王瑛,杨海荣,张莉,等.H2O2和ABA对干旱高温复合胁迫诱导的玉米叶片抗氧化防护基因表达的影响[J].河南农业大学学报,2011,45(06):634-639.
[5]蒋妮,覃柳燕,李力,等.环境胁迫对药用植物次生代谢产物的影响[J].湖北农业科学,2012,51(8):1528-1532.
[6] Anwar S, Iqbal M, Akram H M, et al. Influence of Drought Applied at Different Growth Stages on Kernel Yield and Quality in Maize(Zea mays L.)[J]. Communications in Soil Science and Plant Analysis,2016,47(19/22):2225-2232.
[7]白英俊,李国瑞,黄凤兰,等.活性氧与植物抗氧化系统研究进展[J].安徽农业科学,2017,45(36):1-3.
[8] Yang J, Kilmartin P A, Mingtao F, et al. Assessment of phenolic contributors to antioxidant activity of new kiwifruit cultivars using cyclic voltammetry combined with HPLC[J]. Food Chemistry, 2018,268:77-85.
[9] Ghahfarokhi M G, Mansurifar S, Taghizadehmehrjardi R,et al.Effects of drought stress and rewatering on antioxidant systems and relative water content in different growth stages of maize(Zea mays L.)hybrids[J]. Archives of Agronomy and Soil Science,2015,61(4/6):493-506.
[10] Anjum S A, Umair A, Mohsin T, et al. Drought Induced Changes in Growth, Osmolyte Accumulation and Antioxidant Metabolism of Three Maize Hybrids[J]. Frontiers in Plant Science,2017,8:69.
[11] Sarker U, Oba S. Drought stress enhances nutritional and bioactive compounds, phenolic acids and antioxidant capacity of Amaranthus leafy vegetable[J]. BMC Plant Biology,2018,18(1):258.
[12] Sun C X, Li M Q, Gao X X, et al. Metabolic response of maize plants to multifactorrial abiotic stresses[J]. Plant Biology,2016,18(suppl.1):120-129.
[13]成锴,苏晓慧,栗建枝,等.PEG-6000胁迫下玉米品种萌发期抗旱性鉴定与评价[J].玉米科学,2017,25(5):85-90.
[14]刘海芳,魏东伟,刘全军,等.太行菊不同器官中绿原酸和4种黄酮类物质含量研究[J].天然产物研究与开发,2013,25(5):646-651;671.
[15]谢昕.玉米苞叶多酚提取工艺优化[J].农产品加工,2018(7):47-51.
[16]苏贻娟,冯远娇,罗赐君,等.高效液相色谱法测定玉米叶片中的酚酸类化合物[J].玉米科学,2009,17(5):166-168.
[17]赵霞,刘诗慧,张国方,等.16个玉米杂交种的抗旱性评价[J].河南农业科学,2017,46(12):24-28.
[18]王娟,李德全,谷令坤.不同抗旱性玉米幼苗根系抗氧化系统对水分胁迫的反应[J].西北植物学报,2002(2):77-82.
[19]曾智,陈根洪,滕树锐,等.干旱胁迫下厚朴幼苗的生理指标及次生代谢产物对保水剂的响应[J].福建农林大学学报:自然科学版,2016,45(5):544-550.
[20]赵贝贝,叶蕴灵,王莉,等.银杏类黄酮响应非生物胁迫研究进展[J].扬州大学学报:农业与生命科学版,2018,39(3):106-112.
[21]马惠,王琦,赵鸣,等.非生物胁迫对棉花次生代谢及棉蚜种群消长的影响[J].棉花学报,2016,28(4):324-330.
[22] Hodaei M, Rahimmalek M, Arzani A, et al. The effect of water stress on phytochemical accumulation, bioactivecompounds and expression of key genes involved in flavonoid biosynthesisin Chrysanthemum morifolium L.[J]. Industrial Crops&Products,2018,120:295-304.
[23]杨婷.高效液相色谱法在中药鉴定中的应用分析及前景[J].中国化工贸易,2018,10(32):141.
[24]李颖畅,张明辉,马春颖,等.响应面法优化超声波辅助提取蓝莓叶多酚[J].食品工业科技,2015,36(15):234-238.