UPLC法同时快速测定湿地土壤中的五种低分子量有机酸
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摘要
研究了超高效液相色谱(UPLC)法同时测定湿地土壤中柠檬酸、草酸、苹果酸、酒石酸和丁二酸等5种有机酸的方法,并测定了武昌湖、巢湖和鄱阳湖湿地土壤样品中的有机酸组成含量,为揭示有机酸在土壤-植物-根际环境中的转运、分配和分泌提供理论依据。结果表明:利用UPLC法可以快速地分离5种有机酸,比HPLC分离时间缩短1倍左右;各有机酸的加标回收率为99. 17%～103. 55%,相对标准偏差为0. 48%～1. 92%,检测限为0. 05～0. 18 mg/kg,该方法线性关系优良(R~2>0. 997);除了鄱阳湖湿地土壤样品的酒石酸未能检测出,其他湿地土壤样品中的5种有机酸都能检测出;不同湿地土壤样品的优势有机酸种类不同,武昌湖湿地土壤中的柠檬酸和苹果酸对有机酸总量贡献较大,巢湖湿地土壤中的柠檬酸对有机酸总量的贡献最大,鄱阳湖湿地土壤中丁二酸占有机酸总量的比例小于10%;湿地土壤有机酸含量多集中在土壤深度为30～50 cm处,这与该区域植物根系分布及微生物活性较高有关。
A method of fast and simultaneous measuring concentrations of citric acid,oxalic acid,malic acid,tartaric acid and succinic acid in wetland soils using the ultra-performance liquid chromatography was established,and characteristics of organic acids in wetland soils of Wuchanghu lake,Chaohu lake and Poyanghu lake were determined to better reveal transfer,allocation and exudate of organic acids in the soil-plant-rhizosphere. The results showed that the five organic acids can be separated successfully by UPLC and the separation time was about a half of that of HPLC. The recovery rate,relative standard deviation and detection limit for target organic acids were 99. 17%-103. 55%,0. 48%-1. 92% and 0. 05-0. 18 mg/kg,respectively,and the linear fitting coefficients( R~2) were greater than 0. 997. All of the five organic acids were determined in wetland soils,except tartaric acid,which wasn't detected in soils from the Poyanghu lake. The contents of organic acids in different wetland soils varied,citric and malic acids were the dominant acids in soils of the Wuchanghu lake,the content of citric acid was maximum in soils of the Chaohu lake,the percentage of succinic acid to the total amount of organic acids was lower than 10% for soils from the Poyanghu lake. The highest contents of organic acids were found in soils obtained from the depth of 30-50 cm along soil profiles,which may relate to dense distribution of root and high activity of microbial activity at this depth.
A method of fast and simultaneous measuring concentrations of citric acid,oxalic acid,malic acid,tartaric acid and succinic acid in wetland soils using the ultra-performance liquid chromatography was established,and characteristics of organic acids in wetland soils of Wuchanghu lake,Chaohu lake and Poyanghu lake were determined to better reveal transfer,allocation and exudate of organic acids in the soil-plant-rhizosphere. The results showed that the five organic acids can be separated successfully by UPLC and the separation time was about a half of that of HPLC. The recovery rate,relative standard deviation and detection limit for target organic acids were 99. 17%-103. 55%,0. 48%-1. 92% and 0. 05-0. 18 mg/kg,respectively,and the linear fitting coefficients( R~2) were greater than 0. 997. All of the five organic acids were determined in wetland soils,except tartaric acid,which wasn't detected in soils from the Poyanghu lake. The contents of organic acids in different wetland soils varied,citric and malic acids were the dominant acids in soils of the Wuchanghu lake,the content of citric acid was maximum in soils of the Chaohu lake,the percentage of succinic acid to the total amount of organic acids was lower than 10% for soils from the Poyanghu lake. The highest contents of organic acids were found in soils obtained from the depth of 30-50 cm along soil profiles,which may relate to dense distribution of root and high activity of microbial activity at this depth.
引文
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[13]其美,朱静,李敏,等.反相高效液相色谱法测定湿地根际土壤11种有机酸[J].环境科学与技术,2017,40(S1):255-261.
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[15]沈盼盼,谢晓梅,张圣龙,等. UPLC法测定宣木瓜中齐墩果酸、熊果酸[J].南京中医药大学学报,2013,29(3):271-273.
[16]朱粉霞,丁淑敏,陈斌,等UPLC法同时测定脉络宁注射液中8种有机酸[J].中成药,2013,35(7):1449-1452.
[17]孔维松,宋春满,杨叶昆,等.超高效液相色谱法同时测定烟草中非挥发有机酸[J].云南民族大学学报(自然科学版),2018,27(6):455-459.
[18]黄天志,王世杰,刘秀明,等.逐级提取-高效液相色谱法快速测定植物组织中8种有机酸[J].色谱,2014(12):1356-1361.
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[22]杨红,周立祥,杨志敏,等.植物根系分泌物中有机酸的分析方法[J].分析测试学报,2001,20(4):19-22.
[23] Dinkci N,Akalín AS,Gonc S,et al. Isocratic reverse-phase HPLC for determination of organic acids in KargíTulum cheese[J]. Chromatographia,2007,66(S1):45-49.
[24]赵宽,吴沿友. 4种植物幼苗根茎叶及根系分泌物中低分子量有机酸的特征[J].西北植物学报,2014(5):1002-1007.
[25] López-Bucio J,Nieto-jacobo M F,Ramírez-Rodríguez V,et al. Organic acid metabolism in plants:From adaptive physiology to transgenic varieties for cultivation in extreme soils[J]. Plant Science,2000,160(1):1-13.
[26]纪昌品,王华.鄱阳湖湿地植物群落分布特征及其对土壤环境因子的响应[J].生态环境学报,2018,27(8):1424-1431.
[2] Chen Y,Wang Y,Yeh K. Role of root exudates in metal acquisition and tolerance[J]. Current Opinion in Plant Biology,2017,39:66-72.
[3]赵宽,周葆华,马万征,等.不同环境胁迫对根系分泌有机酸的影响研究进展[J].土壤,2016,48(2):235-240.
[4] Jones D L,Nguyen C,Finlay R D. Carbon flow in the rhizosphere:Carbon trading at the soil-root interface[J]. Plant and Soil,2009,321(1-2):5-33.
[5]丁永祯,李志安,邹碧.土壤低分子量有机酸及其生态功能[J].土壤,2005,37(3):243-250.
[6]赵宽,吴沿友.根系分泌的有机酸及其对喀斯特植物、土壤碳汇的影响[J].中国岩溶,2012,30(4):104-109.
[7] Bais H P,Weir T L,Perry L G,et al. The role of root exudates in rhizosphere interactions with plants and other organisms[J]. Annual Review of Plant Biology,2006,57(1):233-266.
[8] Haichar F E Z,Santaella C,Heulin T,et al. Root exudates mediated interactions belowground[J]. Soil Biology&Biochemistry,2014,77(7):69-80.
[9] Ehling S,Cole S. Analysis of organic acids in fruit juices by liquid chromatography-mass spectrometry:An enhanced tool for authenticity testing[J]. Journal of Agricultural and Food Chemistry,2011,59(6):2229-223
[10] Jaitz L,Mueller B,Koellensperger G,et al. LC-MS analysis of low molecular weight organic acids derived from root exudation[J]. Analytical and Bioanalytical Chemistry,2011,400(8):2587-2596.
[11]杨红,黄焕忠,周立祥,等.植物根系分泌物中有机酸的分析方法[J].分析测试学报,2001,20(4):19-21.
[12]韩海峰,王庆,刘霞,等.聚合离子液体为添加剂的毛细管电泳法用于快速高效分离饮料中7种有机酸[J].色谱,2012,30(5):538-542.
[13]其美,朱静,李敏,等.反相高效液相色谱法测定湿地根际土壤11种有机酸[J].环境科学与技术,2017,40(S1):255-261.
[14]胡蓉,乙引,伍庆. RP-HPLC法测定植物根系中有机酸含量[J].贵州农业科学,2007,35(2):20-21.
[15]沈盼盼,谢晓梅,张圣龙,等. UPLC法测定宣木瓜中齐墩果酸、熊果酸[J].南京中医药大学学报,2013,29(3):271-273.
[16]朱粉霞,丁淑敏,陈斌,等UPLC法同时测定脉络宁注射液中8种有机酸[J].中成药,2013,35(7):1449-1452.
[17]孔维松,宋春满,杨叶昆,等.超高效液相色谱法同时测定烟草中非挥发有机酸[J].云南民族大学学报(自然科学版),2018,27(6):455-459.
[18]黄天志,王世杰,刘秀明,等.逐级提取-高效液相色谱法快速测定植物组织中8种有机酸[J].色谱,2014(12):1356-1361.
[19]叶思诚,谭晓风,袁军,等.油茶根系及分泌物中有机酸的HPLC法测定[J].南京林业大学学报(自然科学版),2013,37(6):59-63.
[20] 20] Khosravi F,Rastakhiz N,Iranmanesh N,et al. Determination of organic acids in fruit juices by UPLC[J]. International Journal of Life Sciences,2015,9(5):41-44.
[21]张浩,朱海林,陈金鸾,等. RP-HPLC法测定林下山参片中有机酸的含量[J].人参研究,2016,28(1):6-8.
[22]杨红,周立祥,杨志敏,等.植物根系分泌物中有机酸的分析方法[J].分析测试学报,2001,20(4):19-22.
[23] Dinkci N,Akalín AS,Gonc S,et al. Isocratic reverse-phase HPLC for determination of organic acids in KargíTulum cheese[J]. Chromatographia,2007,66(S1):45-49.
[24]赵宽,吴沿友. 4种植物幼苗根茎叶及根系分泌物中低分子量有机酸的特征[J].西北植物学报,2014(5):1002-1007.
[25] López-Bucio J,Nieto-jacobo M F,Ramírez-Rodríguez V,et al. Organic acid metabolism in plants:From adaptive physiology to transgenic varieties for cultivation in extreme soils[J]. Plant Science,2000,160(1):1-13.
[26]纪昌品,王华.鄱阳湖湿地植物群落分布特征及其对土壤环境因子的响应[J].生态环境学报,2018,27(8):1424-1431.