浊度对活体叶绿素荧光检测的影响及修正方法
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摘要
本文研究了泥沙悬浮液浊度对叶绿素荧光计测定水样中活体叶绿素荧光的影响。结果表明,浊度产生的荧光效应和遮蔽效应对活体叶绿素荧光测定具有显著影响。在100NTU浊度范围内,浊度会导致荧光效应使活体叶绿素荧光值测量结果随着浊度的增大而增大,并构成良好的线性关系;实际测定水体中硅藻的活体叶绿素荧光值时,泥沙悬浮液的浊度会导致遮蔽效应使活体叶绿素荧光值测量结果随着浊度的增大而减小,水体中硅藻质量浓度越高遮蔽效应越显著,在1 000 mV(约10μg/L)硅藻质量浓度,100NTU浊度条件下,浊度遮蔽效应可导致活体叶绿素荧光值偏低20%。根据近海泥沙实验数据,本文提出了一种活体叶绿素荧光检测的浊度影响修正方法,在100NTU浊度内,修正后叶绿素荧光值偏差率都下降到±10%以内,校准后的荧光值与真实叶绿素荧光值较为接近。
The influence of sediment turbidity on chlorophyll sensor determination of in vivo chlorophyll fluorescence was studied.The results show that turbidity can cause fluorescence effect and light shielding effect on the determination of chlorophyll fluorescence.In the range of 100 NTU turbidity,the fluorescence value of chlorophyll shows a good linear relationship with the turbidity of sediment suspension,which increases with the increase of turbidity.When actually measuring the in vivo chlorophyll fluorescence of diatoms in water,turbidity of sediment suspension causes light shielding effect that decreases in vivo chlorophyll fluorescence as turbidity increases.The concentration of microalgae in the water is higher,the light shielding effect is stronger.In 1 000 mV(including 10μg/L),the highest concentration of microalgae,sediment suspension can reduce the chlorophyll fluorescence value by 20% at 100 NTU turbidity.Based on the experimental data,a correction method of turbidity effect of in vivo chlorophyll fluorescence detection is proposed.Within 100 NTU turbidity,the corrected deviation of chlorophyll fluorescence values is reduced to less than ±10% and the corrected fluorescence value is close to the true chlorophyll fluorescence.
The influence of sediment turbidity on chlorophyll sensor determination of in vivo chlorophyll fluorescence was studied.The results show that turbidity can cause fluorescence effect and light shielding effect on the determination of chlorophyll fluorescence.In the range of 100 NTU turbidity,the fluorescence value of chlorophyll shows a good linear relationship with the turbidity of sediment suspension,which increases with the increase of turbidity.When actually measuring the in vivo chlorophyll fluorescence of diatoms in water,turbidity of sediment suspension causes light shielding effect that decreases in vivo chlorophyll fluorescence as turbidity increases.The concentration of microalgae in the water is higher,the light shielding effect is stronger.In 1 000 mV(including 10μg/L),the highest concentration of microalgae,sediment suspension can reduce the chlorophyll fluorescence value by 20% at 100 NTU turbidity.Based on the experimental data,a correction method of turbidity effect of in vivo chlorophyll fluorescence detection is proposed.Within 100 NTU turbidity,the corrected deviation of chlorophyll fluorescence values is reduced to less than ±10% and the corrected fluorescence value is close to the true chlorophyll fluorescence.
引文
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[21]NELSON D M,TRéGUER P,BRZEZINSKI M A,et al.Production and dissolution of biogenic silica in the ocean:Revised global estimates,comparison with regional data and relationship to biogenic sedimentation[J].Global Biogeochemical Cycles,1995,9(3):359-372.
[22]YU Jin-lan.The response of photosynthesis and carbonic anhydrase of Skeletonema costatum to environmental factors[D].Guangzhou:Guangzhou University,2011.余锦兰.中肋骨条藻光合作用和碳酸酐酶对环境因子的响应[D].广州:广州大学,2011.
[23]HU YI,YE Shu-ming,LOU Kai-kai,et al.Design of high precision in-situ detection of chlorophyll-a[J].Chinese Journal of Scientific Instrument,2012,33(6):1 372-1 376.胡轶,叶树明,楼凯凯,等.原位叶绿素a精密测量方案设计[J].仪器仪表学报,2012,33(6):1 372-1 376.
[2]CUI Jian-sheng,LPeng-yi,DUAN Li-li.Development of seawater chlorophyll on-line detector[J].Analytical Instrumentation,2014(1):12-16.崔建升,吕鹏翼,段莉丽.海水叶绿素现场检测仪的研制及性能研究[J].分析仪器,2014(1):12-16.
[3]LIU Jian-hua.Study on the influence factors of fluorescence determination of chlorophyll in the water[D].Shijiazhuang:Hebei University of Science and Technology,2014.刘建华.荧光法测定水体中叶绿素含量影响因素的研究[D].石家庄:河北科技大学,2014.
[4]GB 17378.7—2007The specification for marine monitoring.Part4:Seawater analysis[S].Beijing:Standards Press of China,2008.GB 17378.7—2007海洋监测规范.第4部分:海水分析[S].北京:中国标准出版社出版,2008.
[5]DAI Rong-ji,TONG Bin,HUANG Chun,et al.Determination of chlorophylls in drinking water by HPLC[J].Transactions of Beijing Institute of Technology,2006,26(1):87-89.戴荣继,佟斌,黄春,等.HPLC测定饮用水中藻类叶绿素含量[J].北京理工大学学报,2006,26(1):87-89.
[6]LORENZEN C J.A method for the continuous measurement of in vivo chlorophyll concentration[J].Deep Sea Research&Oceanographic Abstracts,1966,13(2):223-227.
[7]FAN Yan.The research of new method for extraction and determination of chlorophyll ain macro Algae[R].Beijing international conference on military environmental protection,2004.范燕.提取测定大型海藻叶绿素a的新方法研究[R].北京国际军事环保研讨会,2004.
[8]ZHANG Ke-ke,YAN Xing-kui,CHEN Shi-zhe,et al.Design of insitu chlorophyll-asensor by fluorescence method[J].Shangdong Science,2013,26(3):37-40.张可可,闫星魁,陈世哲,等.荧光法海水叶绿素a传感器设计[J].山东科学,2013,26(3):37-40.
[9]ZHANG Jiang-long.The application of chorophyll testing instrumentto on-line water quality automatic monitoring system[J].Modern Scientific Instruments,2007(1):41-43.张江龙.叶绿素监测仪器在水质自动监测应用中的优劣浅析[J].现代科学仪器,2007(1):41-43.
[10]XIA Da-ying,WANG Zhen-xian,XIA Rong-huan,et al.The underwater fluorometer and its application in marine in-situ detection[J].Journal of Oceanography of Huanghai&Bohai Seas,1997,15(2):64-70.夏达英,王振先,夏荣环,等.水中荧光计及其在海洋现场探测中的应用[J].黄渤海海洋,1997,15(2):64-70.
[11]LI Zu-hua,LI Tian-shen,ZHOU Wen-dong.Comparison of automatic monitoring instrument and laboratorial national standard method to detect chl-ain seawater[J].Instrumentation Analysis Monitoring,2013(3):41-43.李祖华,李天深,周文东.海水自动监测站与实验室国标方法测定海水中叶绿素的比对分析[J].仪器仪表与分析监测,2013(3):41-43.
[12]GREGOR J,MARSLEK B.Freshwater phytoplankton quantification by chlorophyll a:A comparative study of in vitro,in vivo and in situ methods[J].Water Research,2004,38(3):517.
[13]MILLS D K,TETT P B.Use of a recording fluorometer for continuous measurement of phytoplankton concentration[J].Proc SPIE:Environment and Pollution Measurement Sensors and Systems,1990,1269(8):106-115.
[14]LPeng-yi,CUI Jian-sheng,DUAN Li-li.Effect of turbidity on fluorescence intensity of chlorophyll-ain water[J].Physical Testing and Chemical Analysis(Part B:Chemical Analysis),2015,51(6):741-744.吕鹏翼,崔建升,段莉丽.浊度对荧光光谱法测定水体中叶绿素a的影响[J].理化检验-化学分册,2015,51(6):741-744.
[15]LIANG Zhou-rui,WANG Fei-jiu,SUN Xiu-tao,et al.Effects of light intensity,temperature and salinity on newborn branches of Sargassum thunbergii evaluated with chlorophyll fluorescence assay[J].Marine Sciences,2011,35(12):21-27.梁洲瑞,王飞久,孙修涛,等.利用叶绿素荧光技术揭示光照、温度和盐度对鼠尾藻嫩芽的影响[J].海洋科学,2011,35(12):21-27.
[16]ZHENG Min-hui.Variation of in vivo fluorescence during diatom growth and its effect on chlorophyll measurement[D].Wuhan:China University of Geosciences,2016.郑旻辉.硅藻生长过程中活体荧光变化规律及其对叶绿素测量的影响[D].武汉:中国地质大学,2016.
[17]GB 13200—91 Water quality-Determination of turbidity[S].Beijing:National Environmental Protection Agency,1992.GB 13200—91水质——浊度的测定[S].北京:国家环境保护局,1992.
[18]HUA Ru-cheng.Culture and utilization of single cell algae[M].Beijing:Agricultural Press,1986.华汝成.单细胞藻类的培养与利用[M].北京:农业出版社,1986.
[19]GUILLARD R R L.Culture of phytoplankton for feeding marine invertebrates[M].Springer US,1975:29-60.
[20]ZHAO Yang-yong,SHEN Bi-jun,HU Jian-lin,et al.Influence of turbidity on chlorophyll aby fluorescence method[C].Academic Annual Meeting of Chinese Society of Environmental Science,2013.赵洋甬,沈碧君,胡建林,等.浊度对荧光法测定叶绿素a的影响[C].中国环境科学学会2013年学术年会,2013.
[21]NELSON D M,TRéGUER P,BRZEZINSKI M A,et al.Production and dissolution of biogenic silica in the ocean:Revised global estimates,comparison with regional data and relationship to biogenic sedimentation[J].Global Biogeochemical Cycles,1995,9(3):359-372.
[22]YU Jin-lan.The response of photosynthesis and carbonic anhydrase of Skeletonema costatum to environmental factors[D].Guangzhou:Guangzhou University,2011.余锦兰.中肋骨条藻光合作用和碳酸酐酶对环境因子的响应[D].广州:广州大学,2011.
[23]HU YI,YE Shu-ming,LOU Kai-kai,et al.Design of high precision in-situ detection of chlorophyll-a[J].Chinese Journal of Scientific Instrument,2012,33(6):1 372-1 376.胡轶,叶树明,楼凯凯,等.原位叶绿素a精密测量方案设计[J].仪器仪表学报,2012,33(6):1 372-1 376.