环境样品中氨基糖分析检测方法的研究进展

doi:10.11867/j.issn.1001-8166.2019.01.0084

地球科学进展 ›› 2019, Vol. 34 ›› Issue (1): 84 -92. doi: 10.11867/j.issn.1001-8166.2019.01.0084

环境样品中氨基糖分析检测方法的研究进展

魏金娥 ¹(

),张洪海 ^1, ²,陈岩 ^1, ^*(

),杨桂朋 ^1, ²

1. 中国海洋大学化学化工学院,山东青岛 266100
2. 青岛海洋科学与技术国家实验室海洋生态与环境科学功能实验室,山东青岛 266237

收稿日期:2018-10-10 修回日期:2018-12-02 出版日期:2019-01-10
通讯作者: 陈岩 E-mail:weijine@stu.ouc.edu.cn;yanchen@ouc.edu.cn
基金资助:
中央高校基本科研业务费专项项目“东海溶解有机物的光化学降解及对活性气体产生机制的影响研究”(编号：201762030)

Progress on Analytical Methods of Amino Sugars in Environmental Samples

Jin’e Wei ¹( ),Honghai Zhang ^1, ²,Yan Chen ^1, ^*( ),Guipeng Yang ^1, ²

1. College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266100, China
2. Function Laboratory of Marine Ecology and Environmental Science, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China

Received:2018-10-10 Revised:2018-12-02 Online:2019-01-10 Published:2019-03-05
Contact: Yan Chen E-mail:weijine@stu.ouc.edu.cn;yanchen@ouc.edu.cn
About author:Wei Jin’e (1995- ), female, Weifang City, Shandong Province, Master student. Research areas include marine biogeochemistry cycles of amino sugars. E-mail: weijine@stu.ouc.edu.cn |Chen Yan (1982- ), male, Zibo City, Shandong Province, Engineer. Research areas include marine biogeochemistry cycles. E-mail: yanchen@ouc.edu.cn
Supported by:
Project supported by the Fundamental Research Funds for the Central Universities “Photochemical degradation of dissolved organic matter in the East China Sea and its influence on the production of active gases”(No.201762030)

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氨基糖作为环境中重要的生源有机质，其含量和组成信息能够反映有机质来源和微生物对有机质的贡献。国内外针对环境中氨基糖的分析测定开展了大量的研究，但系统地评述其分析方法及最新进展的工作仍然较少。从前处理方法和检测技术等方面，系统归纳了气相色谱法、高效液相色谱法和红外光谱法3类常用的氨基糖分析方法的优缺点和适用条件。在前处理方面，气相色谱法前处理过程较为繁琐且需衍生化处理，而高效液相色谱法前处理相对简单且更易实现自动化；在仪器检测方面，气相色谱法能同时检测环境中4种主要的氨基糖（氨基葡萄糖、氨基半乳糖、氨基甘露糖和胞壁酸），并且比高效液相色谱法稳定，而红外光谱法虽具有结构定性方面的优势，但其灵敏度较低。总之，尚未有一种分析方法既能保证氨基糖的灵敏分析又能兼顾实验效率，因此在分析环境样品中氨基糖时应根据样品形态和检测要求合理选择分析方法。在检测分析方面，今后可以在分析方法的经济性、兼容互补性以及在线自动化等方面着力开展工作，为更深入地开展环境中氨基糖生物地球化学过程的研究提供技术支持。

关键词: 氨基糖, 分析方法, 气相色谱, 液相色谱, 红外光谱

As a kind of important biogenic organic matter, amino sugars can effectively provide insights for the source of organic matters and the contribution of bacterial organic matters based on their concentrations and compositions in the environment. A large number of studies on the analysis of amino sugars have been conducted for environmental samples throughout the world. However, comprehensive and systematic reviews of new progress on the analytical method are still rare. From the aspects of pretreatment methods and detection techniques, the advantages and disadvantages and applicable conditions of three common methods (eg. gas chromatography, high performance liquid chromatography and infrared spectroscopy) were systematically summarized. In terms of pretreatment, the process of the gas chromatography is cumbersome and requires derivatization, while the pretreatment of high performance liquid chromatography is relatively simple and easier to automate. In respect of instrument detection, the gas chromatography can detect four amino sugars (glucosamine, galactosamine, mannosamine and muramic acid) simultaneously and is more stable than the high performance liquid chromatography. In addition, the infrared spectroscopy method has the advantages of structural qualitative, however, its sensitivity is lower. There is no analytical method that can guarantee both sensitive analysis of amino sugars and experimental efficiency. Therefore, the analytical method should be reasonably selected according to the form of the sample and the requirements in the analysis of amino sugars. Further work should focus on economy, compatibility and online automation of analytical methods, so as to provide technical support for the research on biogeochemical processes of amino sugars in the environment.

Key words: Amino sugars, Analytical methods, Gas chromatography, Liquid chromatography, Infrared reflectance spectroscopy.

中图分类号:

P593

图1 AS样品的GC方法处理过程

Fig. 1 Preparation scheme of AS samples with GC method

表1 GC-FID、高效液相色谱—荧光检测法 (HPLC-FLD)和高效阴离子交换色谱—安培检测法 (HPAEC-PAD)分析 AS的检测指标比较

Table 1 Comparison of detection indices of AS by GC-FID, High Performance Liquid Chromatography-Fluorescence Detection( HPLC-FLD) and High Performance Anion Exchange Chromatography-Pulsed Amperometric Detection(HPAEC-PAD)

分析方法	检出限	定量限	检测时间/min	RSD/%	分析对象
GC-FID	-	10~20 μg/mL	19.5	<2	土壤^{[ 13 ]}
HPLC-FLD	1.7~9.0 μg/g	4.2~29.9 μg/g	26	2	土壤和植物^{[ 14 ]}
HPAEC-PAD	1~4 nmol/L	-	90	2~11	海水^{[ 15 ]}

表1 GC-FID、高效液相色谱—荧光检测法 (HPLC-FLD)和高效阴离子交换色谱—安培检测法 (HPAEC-PAD)分析 AS的检测指标比较

分析方法	检出限	定量限	检测时间/min	RSD/%	分析对象
GC-FID	-	10~20 μg/mL	19.5	<2	土壤^{[ 13 ]}
HPLC-FLD	1.7~9.0 μg/g	4.2~29.9 μg/g	26	2	土壤和植物^{[ 14 ]}
HPAEC-PAD	1~4 nmol/L	-	90	2~11	海水^{[ 15 ]}

图2 GlcN和MurA糖腈乙酰化衍生方程式（据参考文献[ 16 ]修改）

Fig. 2 Schemes for formation of the aldononitrile acetate derivatized GlcN and MurA (modified after reference [ 16 ])

图3 AS样品的HPAEC-PAD方法处理过程（据参考文献[ 15 ]修改）

Fig. 3 Preparation scheme of AS samples with HPAEC-PAD method (modified after reference [15])

表2 不同分析方法前处理比较

Table 2 Comparison for pretreatment of different analytical methods

分析方法	水解	纯化	衍生状况	应用	参考文献
GC-FID	6 mol/L HCl	调pH、无水甲醇	需要	土壤	[ 13 ]
HPLC-FLD	6 mol/L HCl	无	需要	土壤和根系	[ 14 ]
HPAEC-PAD	3 mol/L HCl	固相萃取柱	不需要	海水	[ 15 ]
HPLC-IRMS	6 mol/L HCl	调pH、无水甲醇、固相萃取柱	不需要	土壤	[ 40 ]
IR	6 mol/L HCl	调pH、无水甲醇	不需要	土壤	[ 45 ]

表2 不同分析方法前处理比较

Table 2 Comparison for pretreatment of different analytical methods

分析方法	水解	纯化	衍生状况	应用	参考文献
GC-FID	6 mol/L HCl	调pH、无水甲醇	需要	土壤	[ 13 ]
HPLC-FLD	6 mol/L HCl	无	需要	土壤和根系	[ 14 ]
HPAEC-PAD	3 mol/L HCl	固相萃取柱	不需要	海水	[ 15 ]
HPLC-IRMS	6 mol/L HCl	调pH、无水甲醇、固相萃取柱	不需要	土壤	[ 40 ]
IR	6 mol/L HCl	调pH、无水甲醇	不需要	土壤	[ 45 ]

表3 AS分析方法比较

分析方法	应用对象	优势	不足	发展趋势
GC-FID	土壤中GlcN，GalN，ManN和MurA	同时定量4种AS	$①$ 样品须经过衍生化处理； $②$ 易受氨基糖苷类抗生素干扰	衍生过程由离线向“在线”发展
GC-IRMS	土壤和海洋沉积物中AS的δ¹³C	受样品中杂质影响较小	$①$ 样品须经过衍生化处理； $②$ 检测丰度高的同位素时，准确度会下降	多同位素同时检测
GC-MS	土壤中AS的APE	不仅能提供同位素比值信息还能提供分子结构信息	$①$ 样品须经过衍生化处理； $②$ 在定量方面有一定的缺陷	向高分辨质谱发展
HPLC-FLD	土壤和根系中GlcN，GalN，ManN和MurA	实现在线衍生化	$①$ 需要衍生化处理； $②$ 受氨基酸干扰强； $③$ 不能同时检测中性糖	发展同时高效检测AS和氨基酸的方法
HPAEC-PAD	海水和沉积物中GlcN，GalN，ManN和MurA	$①$ 无需衍生化前处理； $②$ 干扰较小	$①$ MurA不能与其他3种AS同时分析； $②$ 易受CO₂影响	前处理过程向自动化发展
HPLC-IRMS	土壤中AS的δ¹³C	无需衍生化前处理	$①$ 对CO₂污染敏感； $②$ MurA不能与其他3种AS同时分析	多同位素同时检测
UPLC-HRMS	土壤中游离GlcN和MurA	$①$ 无需衍生化前处理； $②$ 仪器灵敏度高、检测速度快	$①$ 仪器使用及维护成本高； $②$ 仪器需要专业的人员进行维护操作	向多维液相色谱发展
IR	土壤中TAS-N，GlcN，GalN和MurA	$①$ 无危险试剂使用； $②$ 检测快速； $③$ 与色谱方法相比，成本低	相对于色谱方法，方法的灵敏度不高	向高空间分辨率的组分分析方向发展

表3 AS分析方法比较

分析方法	应用对象	优势	不足	发展趋势
GC-FID	土壤中GlcN，GalN，ManN和MurA	同时定量4种AS	$①$ 样品须经过衍生化处理； $②$ 易受氨基糖苷类抗生素干扰	衍生过程由离线向“在线”发展
GC-IRMS	土壤和海洋沉积物中AS的δ¹³C	受样品中杂质影响较小	$①$ 样品须经过衍生化处理； $②$ 检测丰度高的同位素时，准确度会下降	多同位素同时检测
GC-MS	土壤中AS的APE	不仅能提供同位素比值信息还能提供分子结构信息	$①$ 样品须经过衍生化处理； $②$ 在定量方面有一定的缺陷	向高分辨质谱发展
HPLC-FLD	土壤和根系中GlcN，GalN，ManN和MurA	实现在线衍生化	$①$ 需要衍生化处理； $②$ 受氨基酸干扰强； $③$ 不能同时检测中性糖	发展同时高效检测AS和氨基酸的方法
HPAEC-PAD	海水和沉积物中GlcN，GalN，ManN和MurA	$①$ 无需衍生化前处理； $②$ 干扰较小	$①$ MurA不能与其他3种AS同时分析； $②$ 易受CO₂影响	前处理过程向自动化发展
HPLC-IRMS	土壤中AS的δ¹³C	无需衍生化前处理	$①$ 对CO₂污染敏感； $②$ MurA不能与其他3种AS同时分析	多同位素同时检测
UPLC-HRMS	土壤中游离GlcN和MurA	$①$ 无需衍生化前处理； $②$ 仪器灵敏度高、检测速度快	$①$ 仪器使用及维护成本高； $②$ 仪器需要专业的人员进行维护操作	向多维液相色谱发展
IR	土壤中TAS-N，GlcN，GalN和MurA	$①$ 无危险试剂使用； $②$ 检测快速； $③$ 与色谱方法相比，成本低	相对于色谱方法，方法的灵敏度不高	向高空间分辨率的组分分析方向发展

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