海中有色可溶有机物荧光光谱现场探测技术研究
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摘要
海水中有色可溶有机物(Chromophoric Dissolved Organic Matter:CDOM,国内通常称之为黄色物质)的研究在全球碳循环、海洋水色、海洋生态、海洋动力环境和海洋污染评价等研究领域具有重要的意义。荧光光谱具有灵敏度高、选择性好等优点,且不需要对目标物进行分离与处理,近年来已被广泛应用于水体中CDOM的检测和分析,但仍以采集水样的实验室分析方法为主。本论文针对CDOM现场探测技术展开研究,具体工作如下：
1、研制了一套小型化的、基于ROV的CDOM水下原位荧光光谱探测系统。该系统采用355nn紫外波长激光激发,微型光纤光谱仪分光探测,可在水下原位实时获得黄色物质的整条光谱曲线。系统采用ROV平台和荧光光谱探测装置一体化设计,先后进行了实验室密封实验、水池实验、海上实验,工作稳定,与紫外-可见分光光度计对比试验,也具有较好相关性(R=0.67)
2、提出了一种实用的激光/LED混合三波长同时激发荧光的测量方法,并成功进行了实验室原理验证。采用266nm/532nm双波长激光器和375nmLED作为激发光源,可获得较为全面的水下荧光信号,包括类蛋白CDOM、类腐殖质CDOM、叶绿素等。
3、本文利用ICCD开展了CDOM时间分辨荧光光谱技术研究。分别利用355nm和266nm激光器对CDOM进行激发,类色氨酸和两种类腐殖质的荧光寿命均小于10ns,但有所差别,利用荧光寿命不同可对成分进行区分。另外,完成了一套基于ICCD的微脉冲海洋荧光激光雷达系统原理验证,并以此为基础计算了机载海洋荧光激光雷达采用ICCD作为探测器的可行性。
4、开展了荧光-吸收光谱联合探测技术研究,利用液芯光纤作为样品池对吸收进行高灵敏度测量,同时对CDOM荧光光谱进行现场定标,经过实验室对比实验,荧光强度和吸收系数具有较好的相关性(R=0.98)
5、针对CDOM物质成分分析,从三方面开展了荧光光谱-拉曼光谱联合探测技术研究：深紫外荧光光谱与常规可见光激发拉曼光谱的联合、深紫外荧光光谱与紫外共振拉曼光谱的联合,深紫外荧光光谱与表面增强拉曼光谱的联合,其中第三种联合最为理想。成功地利用一套装置同时获得了CDOM的荧光光谱和表面增强拉曼光谱。
The research on chromophoric dissolved organic matter (CDOM) is very important for ocean color, global carbon cycle, oceanic ecology, oceanic dynamical environments, and pollution monitoring in ocean. CDOM measurements with fluorescence techniques have high sensitivity and selectivity, and can operate simply without water sample filter and pretreatment. Fluorescence spectroscopy has been used in CDOM research and measurements widely in recent years. While most fluorescence research at the present time needs to collect water sample and measure spectra with fluorospectrophotometer in lab. So the development of in-situ fluorescence devices and techniques has great utility value and necessity. This thesis focuses on the CDOM in-situ fluorescence techniques and devices. The main contents are as follows:
1. A compact underwater CDOM in-situ fluorescence prototype has been successfully developed based on ROV platform. This prototype can get CDOM spectra curves in real time with a355nm UV laser and a small optical fiber spectrometer. The main feature of this prototype is the integration configuration design between ROV platform and in-situ fluorescence device chamber. Series performance tests such as laboratory waterproof tests, swimming pool tests, and oceanic field deployments have demonstrated the utility of the prototype as an integrated tool for research and observations. The fluorescence intensity ratio has good correlation with absorption coefficient measured with UV-VIS spectrophotometry.
2. A novel multi-wavelength fluorescence measurement method was proposed and verification experiment proved its fesibility. This method provides fluorescence spectra for assessment of CDOM, chlorophyll a, and pigments with combined dual-wavelength (266nm and532nm) laser and375nm LED as excitation source.
3. Time resolved fluorescence spectroscopy has been used for CDOM research with ICCD and pulsed laser excited at355nm or266nm. When excited at266nm, three main components with different fluorescence lifetime have been differentiated successfully. Furthermore, a compact ocean lidar verification device was performed with ICCD and a micro-pulsed laser.
4. Fluorescence and absorption spectroscopy joint analysis has been performed with a liquid core optical fiber used as absorption sample cell. The absorption measurement with a liquid core optical fiber can improve sensitivity by increasing optical path, and can also calibrate fluorescence data in field deployments. The results show good correlation(R=0.98)between fluorescence intensity ratio and absorption in laboratory experiments.
5、Fluorescence and Raman spectroscopy joint research has been performed for CDOM composition analysis. Three combination modes has been studied including deep UV fluorescence and VIS Raman joint, deep UV fluorescence and UV resonance Raman joint, deep UV fluorescence and surface enhanced resonance Raman joint. Between the three combination modes, deep UV fluorescence and surface enhanced resonance Raman joint analysis is the most satisfactory. Fluorescence and surface enhanced resonance Raman spectra were obtained simultaneously with the same device.
1、研制了一套小型化的、基于ROV的CDOM水下原位荧光光谱探测系统。该系统采用355nn紫外波长激光激发,微型光纤光谱仪分光探测,可在水下原位实时获得黄色物质的整条光谱曲线。系统采用ROV平台和荧光光谱探测装置一体化设计,先后进行了实验室密封实验、水池实验、海上实验,工作稳定,与紫外-可见分光光度计对比试验,也具有较好相关性(R=0.67)
2、提出了一种实用的激光/LED混合三波长同时激发荧光的测量方法,并成功进行了实验室原理验证。采用266nm/532nm双波长激光器和375nmLED作为激发光源,可获得较为全面的水下荧光信号,包括类蛋白CDOM、类腐殖质CDOM、叶绿素等。
3、本文利用ICCD开展了CDOM时间分辨荧光光谱技术研究。分别利用355nm和266nm激光器对CDOM进行激发,类色氨酸和两种类腐殖质的荧光寿命均小于10ns,但有所差别,利用荧光寿命不同可对成分进行区分。另外,完成了一套基于ICCD的微脉冲海洋荧光激光雷达系统原理验证,并以此为基础计算了机载海洋荧光激光雷达采用ICCD作为探测器的可行性。
4、开展了荧光-吸收光谱联合探测技术研究,利用液芯光纤作为样品池对吸收进行高灵敏度测量,同时对CDOM荧光光谱进行现场定标,经过实验室对比实验,荧光强度和吸收系数具有较好的相关性(R=0.98)
5、针对CDOM物质成分分析,从三方面开展了荧光光谱-拉曼光谱联合探测技术研究：深紫外荧光光谱与常规可见光激发拉曼光谱的联合、深紫外荧光光谱与紫外共振拉曼光谱的联合,深紫外荧光光谱与表面增强拉曼光谱的联合,其中第三种联合最为理想。成功地利用一套装置同时获得了CDOM的荧光光谱和表面增强拉曼光谱。
The research on chromophoric dissolved organic matter (CDOM) is very important for ocean color, global carbon cycle, oceanic ecology, oceanic dynamical environments, and pollution monitoring in ocean. CDOM measurements with fluorescence techniques have high sensitivity and selectivity, and can operate simply without water sample filter and pretreatment. Fluorescence spectroscopy has been used in CDOM research and measurements widely in recent years. While most fluorescence research at the present time needs to collect water sample and measure spectra with fluorospectrophotometer in lab. So the development of in-situ fluorescence devices and techniques has great utility value and necessity. This thesis focuses on the CDOM in-situ fluorescence techniques and devices. The main contents are as follows:
1. A compact underwater CDOM in-situ fluorescence prototype has been successfully developed based on ROV platform. This prototype can get CDOM spectra curves in real time with a355nm UV laser and a small optical fiber spectrometer. The main feature of this prototype is the integration configuration design between ROV platform and in-situ fluorescence device chamber. Series performance tests such as laboratory waterproof tests, swimming pool tests, and oceanic field deployments have demonstrated the utility of the prototype as an integrated tool for research and observations. The fluorescence intensity ratio has good correlation with absorption coefficient measured with UV-VIS spectrophotometry.
2. A novel multi-wavelength fluorescence measurement method was proposed and verification experiment proved its fesibility. This method provides fluorescence spectra for assessment of CDOM, chlorophyll a, and pigments with combined dual-wavelength (266nm and532nm) laser and375nm LED as excitation source.
3. Time resolved fluorescence spectroscopy has been used for CDOM research with ICCD and pulsed laser excited at355nm or266nm. When excited at266nm, three main components with different fluorescence lifetime have been differentiated successfully. Furthermore, a compact ocean lidar verification device was performed with ICCD and a micro-pulsed laser.
4. Fluorescence and absorption spectroscopy joint analysis has been performed with a liquid core optical fiber used as absorption sample cell. The absorption measurement with a liquid core optical fiber can improve sensitivity by increasing optical path, and can also calibrate fluorescence data in field deployments. The results show good correlation(R=0.98)between fluorescence intensity ratio and absorption in laboratory experiments.
5、Fluorescence and Raman spectroscopy joint research has been performed for CDOM composition analysis. Three combination modes has been studied including deep UV fluorescence and VIS Raman joint, deep UV fluorescence and UV resonance Raman joint, deep UV fluorescence and surface enhanced resonance Raman joint. Between the three combination modes, deep UV fluorescence and surface enhanced resonance Raman joint analysis is the most satisfactory. Fluorescence and surface enhanced resonance Raman spectra were obtained simultaneously with the same device.
引文
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