摘要
为提高海水养殖尾水中总磷(Total phosphorus,TP)的检测效率,使用酶标仪代替分光光度计进行海水养殖尾水TP的测定,并对TP的消解方法、酶标板类型与加样量等进行优化。结果表明,使用控温加热板消解60 min与高压灭菌器消解30 min的效果无显著差异(P>0.05);使用48孔板加样1 mL,检测的灵敏度优于其他实验组(96孔板200μL,96孔石英板200μL,48孔板200和500μL,24孔板500μL和1 mL),48孔板加样1 mL与96孔板加样200μL,磷酸盐浓度与吸光度的线性相关性最高(R2=0.9999);对养殖尾水TP测定的精确度和准确度验证结果显示,48孔板加样1 mL的相对误差(Relative error,RE)和相对标准偏差(Relative standard deviation,RSD)分别介于-2.97%~1.59%和0.42%~4.06%,对甘油磷酸二钠(β-GLP)和六偏磷酸钠(SHMP)的回收率分别介于98.2%~99.6%和93.4%~97.1%,TP的测定限(Limit of quantification,LOQ)为0.25μmol·L^(-1);96孔板加样200μL的RE和RSD分别介于-14.03%~0.21%和2.63%~14.23%,对β-GLP和SHMP的回收率分别介于94.7%~99.0%和88.9%~97.3%,TP的LOQ为0.55μmol·L^(-1)。在TP浓度介于0.55~6.4μmol·L^(-1)内,两种优化法的准确度和精密度均符合测定要求,可用于海水养殖尾水TP的测定。
In order to improve the efficiency of the determination of total phosphorus(TP)in the mariculture tailwater,we determined the TP content by microplate reader instead of spectrophotometer in mariculture tailwater,and optimized the digestion method of TP,the type of microwell plate and the amount of sample.The results show that there was no significant difference between 60 min of digestion using a temperature-controlled heating plate and 30 min of digestion using an autoclave(P>0.05).The sensitivity of the assay was better than those of the other experimental groups(200μL for 96-well plate,200μL for96-well quartz plate,200 and 500μL for 48-well plate,500μL and 1 mL for 24-well plate)when using a 48-well plate with 1 mL of sample.The linear correlation between phosphate concentration and absorbance was the best(R2=0.9999)for the 48-well plate with 1 mL and the 96-well plate spiked with 200μL.The accuracy and precision validation of TP determination in mariculture tailwater showed that the relative error(RE)and relative standard deviation(RSD)of the 48-well plate spiked with 1 mL were higher than those of the other experimental groups(96-well plate 200μL,96-well quartz plate 200μL,48-well plate 200μL and 500μL,24-well plate 500μL and 1 mL).The recoveries of disodium glycerophosphate(β-GLP)and sodium hexametaphosphate(SHMP)were 98.2%-99.6%and 93.4%-97.1%,respectively,and the limit of quantification(LOQ)of TP were 0.25μmol·L^(-1);the RE and RSD of 96-well ELISA plate spiked with 200μL were-14.03%-0.21%and 2.63%-14.23%,respectively,and the recoveries forβ-GLP and SHM were 94.7%-99.0%and 88.9%-97.3%,respectively,and the LOQ of TP was 0.55μmol·L^(-1).At TP concentration of 0.55-6.4μmol·L^(-1),the accuracy and precision of the two optimized methods meet the determination requirements and can be used for the determination of TP in mariculture tailwater.
作者
刘雁
佟飞
陈丕茂
袁华荣
冯雪
LIU Yan;TONG Fei;CHEN Pimao;YUAN Huarong;FENG Xue(College of Marine Sciences,Shanghai Ocean University,Shanghai 201306,China;South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences/Scientific Observing and Experimental Station of South China Sea Fishery Resources and Environment,Ministry of Agriculture and Rural Affairs/Key Laboratory of Marine Ranching Technology,Chinese Academy of Fishery Sciences/Guangdong Provincial Key Laboratory of Fishery Ecology and Environment/Guangdong Engineering Technology Research Center of Marine Recreational Fishery,Guangzhou 510300,China)
出处
《南方水产科学》
CAS
CSCD
北大核心
2023年第1期58-66,共9页
South China Fisheries Science
基金
广东省重点领域研发计划项目(2020B1111030002-2)
深圳市科技创新计划(JCYJ20160331141759795)。
关键词
海水养殖尾水
总磷
消解
酶标仪
Mariculture tailwater
Total phosphorus
Digestion
Microplate reader
