自组装膜进样质谱系统及其在砂质沉积物异化硝酸盐还原研究中的应用

谢成军; 宋国栋; 刘素美; 唐继尧; 张桂玲

doi:10.3969/j.issn.0253-4193.2020.02.003

自组装膜进样质谱系统及其在砂质沉积物异化硝酸盐还原研究中的应用

doi: 10.3969/j.issn.0253-4193.2020.02.003

谢成军^{1, 2, 3,},
宋国栋^{1, 2, ,},
刘素美^{1, 2},
唐继尧^{1, 2, 3},
张桂玲^{1, 2}

1.
中国海洋大学海洋化学理论与工程技术教育部重点实验室/海洋高等研究院，山东青岛 266100
2.
青岛海洋科学与技术试点国家实验室海洋生态与环境科学功能实验室，山东青岛 266237
3.
中国海洋大学化学化工学院，山东青岛 266100

基金项目: 国家自然科学基金（41606093，U1806211）；国家重点研发计划（2016YFA0600902，2016YFA0601302）；青岛海洋科学与技术试点国家海洋生态与环境科学功能实验室青年人才培育项目（LMEES-YTSP-2018-02-04）；泰山学者工程专项经费资助。

详细信息

作者简介:
谢成军(1994—)，男，重庆市忠县人，主要从事海洋生物地球化学研究。E-mail：1763462648@qq.com

通讯作者:
宋国栋，男，副教授，主要从事海洋生物地球化学研究。E-mail：gsong@ouc.edu.cn

中图分类号: X52
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出版历程
- 收稿日期: 2019-04-05
- 修回日期: 2019-05-30
- 网络出版日期: 2020-11-18
- 刊出日期: 2020-02-25

Self-assembled membrane injection mass spectrometry system and its application on the study of dissimilatory nitrate reduction in sandy sediments

Xie Chengjun^{1, 2, 3
,},
Song Guodong^{1, 2
, ,},
Liu Sumei^{1, 2},
Tang Jiyao^{1, 2, 3},
Zhang Guiling^{1, 2}

1.
Key Laboratory of Marine Chemistry Theory and Technology, Ministry of Education/Institute for Advanced Ocean Study, Ocean University of China, Qingdao 266100, China
2.
Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China
3.
College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266100, China

摘要

摘要: 沉积物中的异化硝酸盐还原过程对于海洋氮循环起着至关重要的作用。基于¹⁵N标记的培养技术是目前测定沉积物异化硝酸盐还原的主要手段。准确快速测定¹⁵N标记的产物(²⁹N₂、 ³⁰N₂)是量化异化硝酸盐还原各个过程速率的关键。本研究自行组装膜进样质谱系统用于²⁹N₂和³⁰N₂的测定，对其测量条件进行了优化。结果表明，进样蠕动泵进样流速0.80 mL/min，进样时间3～3.5 min，恒温槽温度20～25°C，同时铜还原炉温度在300～600°C的条件下，²⁹N₂/²⁸N₂和³⁰N₂/²⁸N₂的测试精密度分别可以控制在0.1%和1%以内，比较适合²⁹N₂和³⁰N₂的测定。利用自组装的膜进样质谱系统结合¹⁵N标记的培养技术研究了青岛石老人沙滩沉积物中的异化硝酸盐还原过程。石老人沙滩沉积物不存在将硝酸盐完全还原为氮气好氧的反硝化。厌氧铵氧化、厌氧反硝化和异化硝酸盐还原为铵(Dissimilatory Nitrate Reduction to Ammonium, DNRA)的潜在速率 (以湿沉积物N计)分别为(0.05±0.01) nmol /(cm³·h)，(2.32±0.21) nmol /(cm³·h)和(1.02±0.15) nmol /(cm³·h)。厌氧反硝化是硝酸盐异化还原主要的贡献者，其比例接近70%，其次是DNRA，比例可达30%，而厌氧铵氧化的贡献最低，仅为1%。在N₂产生过程中，主要贡献者是反硝化，厌氧铵氧化的贡献仅为2%。
- 膜进样质谱 /
- 砂质沉积物 /
- 反硝化 /
- 厌氧铵氧化 /
- 异化硝酸盐还原为铵
Abstract: Dissimilatory nitrate reduction processes in sediments play a crucial role in marine nitrogen cycle. The most popular method to determine the rates of different dissimilatory nitrate reduction processes is the ¹⁵N labeled technique. Therefore, accurate and rapid determination the concentration of ¹⁵N-labeled products, such as ²⁹N₂ and ³⁰N₂, is the key to quantify the rate of each dissimilatory nitrate reduction process. In this study, we set up a membrane injection mass spectrometry (MIMS) and optimize the operating condition of the MIMS for the determination of ²⁹N₂ and ³⁰N₂. The optimization experiment results indicated that, when the peristaltic pump for sampling flow rate is 0.80 mL/min, the sampling time is 3～3.5 min, the thermostat water bath temperature is 20～25℃, and the copper reduction furnace temperature is 300～600℃, the precision (expressed in coefficient of variation) of the measured ²⁹N₂/²⁸N₂ and ³⁰N₂/²⁸N₂ can be controlled less than 0.1% and 1%, respectively. We used the self-assembled MIMS and combined the ¹⁵N labeling technique to study the dissimilatory nitrate reduction processes in the sandy sediment of the Shilaoren beach in Qingdao. There is no significant aerobic denitrification in the Shilaoren sand that can completely reduce nitrate to N₂. The potential rates of anammox, anaerobic denitrification and dissimilatory nitrate reduction to ammonium (DNRA) are (0.05±0.01) nmol/(cm³·h), (2.32±0.21) nmol/(cm³·h) and (1.02±0.15) nmol/(cm³·h) (N, wet sed.), respectively. Anaerobic denitrification is the major contributor to nitrate dissimilatory reduction, with a ratio of nearly 70%, followed by DNRA, with a ratio of up to 30%, while anammox has the lowest contribution of only 1%. In the N₂ production, the main contributor is anaerobic denitrification, and the contribution of anammox is only 2%.
- membrane injection mass spectrometry (MIMS) /
- sandy sediment /
- denitrification /
- anammox /
- dissimilatory nitrate reduction to ammonium

HTML全文

图 1 自组装膜进样质谱系统示意图

从左到右的主要部件依次为：样品瓶(S)，进样蠕动泵(P)，恒温水浴槽(T)，标准样品制作系统(R-E)，膜进样器(M)，废液回收瓶(W)，冷阱(C)，铜还原炉(F，内含一根装有还原铜丝的石英管)，抽真空系统(V)，离子源(I)，四极杆质量分析器(Q)，二次电子倍增检测器(SEM)，数据处理系统(D)

Fig. 1 Schematic diagram of self-assembled membrane injection mass spectrometry system

The main components from left to right are: sample vial (S), injection peristaltic pump (P), constant temperature water bath (T), standard sample preparation system (R-E), membrane injector (M), waste recovery bottle (W), cold trap (C), copper reduction furnace (F, containing a quartz tube with reduced copper wire), vacuum system (V), Ion source (I), quadrupole mass analyzer (Q), secondary electron multiplier(SEM), data processing system (D)

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图 2 蠕动泵进样流速与进样时间对²⁸N₂信号值的影响

Fig. 2 Effect of peristaltic pump flow speed and injection time on the ²⁸N₂ signal value

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图 3 蠕动泵进样流速和进样时间对²⁹N₂/²⁸N₂和³⁰N₂/²⁸N₂的影响

Fig. 3 Effect of peristaltic pump flow speed and injection time on ²⁹N₂/²⁸N₂ and ³⁰N₂/²⁸N₂

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图 4 不同进样温度下N₂的信号值和N₂/Ar的变异系数

Fig. 4 Signal value of N₂ and coefficient of variation of N₂/Ar at different temperatures

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图 5 铜还原炉温度对于氧气移除效率和N₂同位素测定信号的影响

Fig. 5 Effect of copper reduction furnace temperature on the oxygen removal efficiency and the signal of nitrogen isotope

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图 6 厌氧条件下石老人沙滩沉积物添加¹⁵N培养体系中²⁹N₂、³⁰N₂和$ ^{15}{\rm{NH}}_4^ + $的变化

a.添加$ ^{15}{\rm{NH}}_4^ + $，b.添加$ ^{15}{\rm{NH}}_4^ + $+$^{14}{\rm{NO}}_3^ - $，c. 添加$^{15}{\rm{NO}}_3^ - $，d.添加$^{15}{\rm{NO}}_3^ - $

Fig. 6 Changes of ²⁹N₂, ³⁰N₂ and $ ^{15}{\rm{NH}}_4^ + $ in ¹⁵N culture system of sediment in Shilaoren Beach under anaerobic conditions

a. Added $ ^{15}{\rm{NH}}_4^ + $, b. added $ ^{15}{\rm{NH}}_4^ + $+$^{14}{\rm{NO}}_3^ - $, c. added $^{15}{\rm{NO}}_3^ - $, d. added $^{15}{\rm{NO}}_3^ - $

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图 7 有氧条件下石老人沙滩沉积物添加$^{15}{\rm{NO}}_3^ - $培养体系中DO、²⁹N₂和³⁰N₂的变化

Fig. 7 Changes of DO, ²⁹N₂ and ³⁰N₂ in $^{15}{\rm{NO}}_3^ - $ culture system of sediment in Shilaoren Beach under aerobic conditions

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