Assessment of phytoplankton class abundance using fluorescence excitation-emission matrix by parallel factor analysis and nonnegative least squares

The feasibility of using fluorescence excitation-emission matrix(EEM) along with parallel factor analysis(PARAFAC) and nonnegative least squares(NNLS) method for the differentiation of phytoplankton taxonomic groups was investigated. Forty-one phytoplankton species belonging to 28 genera of five divisions were studied. First, the PARAFAC model was applied to EEMs, and 15 fluorescence components were generated. Second, 15 fluorescence components were found to have a strong discriminating capability based on Bayesian discriminant analysis(BDA). Third, all spectra of the fluorescence component compositions for the 41 phytoplankton species were spectrographically sorted into 61 reference spectra using hierarchical cluster analysis(HCA), and then, the reference spectra were used to establish a database. Finally, the phytoplankton taxonomic groups was differentiated by the reference spectra database using the NNLS method. The five phytoplankton groups were differentiated with the correct discrimination ratios(CDRs) of 100% for single-species samples at the division level. The CDRs for the mixtures were above 91% for the dominant phytoplankton species and above 73% for the subdominant phytoplankton species. Sixteen of the 85 field samples collected from the Changjiang River estuary were analyzed by both HPLC-CHEMTAX and the fluorometric technique developed. The results of both methods reveal that Bacillariophyta was the dominant algal group in these 16 samples and that the subdominant algal groups comprised Dinophyta, Chlorophyta and Cryptophyta. The differentiation results by the fluorometric technique were in good agreement with those from HPLC-CHEMTAX. The results indicate that the fluorometric technique could differentiate algal taxonomic groups accurately at the division level.

Fluorescence Excitation-Emission Matrix Characterization of a Commercial Humic Acid

Excitation-emission matrix fluorescence spectroscopy (EEM) has been widely used to elucidate the origin and structure of humic substances in natural environments. Due to its high sensitivity, good selectivity and non-destructive advantage, the EEM was applied to characterizing a commercial Fluka humic acid (FHA). The results showed that the EEMs of FHA has several Ex/Em peaks. Ionic strength (0- 0.05 mol/L KClO 4) exerted little effect on the fluorescence properties of FHA, while the concentrations (5-100 mg/L) of FHA and pH (2-12) had significant effects. A red shift in the longer wavelength peak region was observed when the concentrations or pH values increased. The fluorescence intensity increased with increasing pH, but slightly decreased in the case of pH= 5.0. The protonation constants (lgK’ HL) of peak B were calculated to be 3.57 and 3.13, indicating that peak B was due to carboxyl groups. The r (A/B) values range from 0.61 to 2.59. A strong linear relationship between r (A/B) and pH was also observed. This indicates that the fluorescence peaks A and B posses similar inherent fluorescence characteristics.

Autofluorescence excitation-emission matrices for diagnosis of colonic cancer

AIM: To investigate the autofluorescence spectroscopic differences in normal and adenomatous coionic tissues and to determine the optimal excitation wavelengths for subsequent study and clinical application. METHODS: Normal and adenomatous coionic tissues were obtained from patients during surgery. A FL/FS920 combined TCSPC spectrofluorimeter and a lifetime spectrometer system were used for fluorescence measurement. Fluorescence excitation wavelengths varying from 260 to 540 nm were used to induce the autofluorescence spectra, and the corresponding emission spectra were recorded from a range starting 20 nm above the excitation wavelength and extending to 800 nm. Emission spectra were assembled into a three-dimensional fluorescence spectroscopy and an excitation-emission matrix (EEM) to exploit endogenous fluorophores and diagnostic information. Then emission spectra of normal and adenomatous coionic tissues at certain excitation wavelengths were compared to determine the optimal excitation wavelengths for diagnosis of coionic cancer. RESULTS: When compared to normal tissues, low NAD (P)H and FAD, but high amino acids and endogenous phorphyrins of protoporphyrin IX characterized the high-grade malignant coionic tissues. The optimal excitation wavelengths for diagnosis of coionic cancer were about 340, 380, 460, and 540 nm. CONCLUSION: Significant differences in autofluorescence peaks and its intensities can be observed in normal and adenomatous coionic tissues. Autofluorescence EEMs are able to identify coionic tissues.

Fluorescence Properties and Chemical Composition of Fine Particles in the Background Atmosphere of North China

To understand the aerosol characteristics in a regional background environment,fine-particle(PM_(2.5),n=228)samples were collected over a one-year period at the Shangdianzi(SDZ)station,which is a Global Atmospheric Watch regional background station in North China.The chemical and optical characteristics of PM_(2.5)were analyzed,including organic carbon,elemental carbon,water-soluble organic carbon,water-soluble inorganic ions,and fluorescent components of watersoluble organic matter.The source factors of major aerosol components are apportioned,and the sources of the fluorescent chromophores are further analyzed.The major chemical components of PM_(2.5)at SDZ were NO_(3)^(-),organic matter,SO_(4)^(2-),and NH_(4)^(+).Annually,water-soluble organic carbon contributed 48%±15%to the total organic carbon.Secondary formation(52%)and fossil fuel combustion(63%)are the largest sources of water-soluble organic matter and water-insoluble organic matter,respectively.In addition,three humic-like and one protein-like matter were identified via parallel factor analysis for excitation–emission matrices.The fluorescence intensities of the components were highest in winter and lowest in summer,indicating the main impact of burning sources.This study contributes to understanding the chemical and optical characteristics of ambient aerosols in the background atmosphere.

Highly sensitive fluorescence quantification of irinotecan in biological fluids with the aid of second-order advantage

A rapid,green and highly sensitive excitation-emission matrix（EEM） fluorescence method was proposed for analysis of irinotecan（CPT-11） in biological fluids including human plasma and urine samples of uncalibrated interferences with the aid of second-order advantage.Due to the serious spectral overlapping from biological matrices,the parallel factor analysis（PARAFAC） and the alternating normalization-weighted error（ANWE） have been recommended to perform directly calibration and overcome the problem which makes the traditional fluorospectrophotometer in trouble.Satisfactory results can be achieved.Furthermore, performance of the proposed method was evaluated based on figures of merit and some statistical parameters.The accuracy of both algorithms was validated by the elliptical joint confidence region（EJCR） test.The precision and repeatability were also investigated by the relative standard deviations（RSDs） of intra-day and inter-day.

Investigation of seasonal variability of CDOM fluorescence in the southern Changjiang River Estuary by EEM-PARAFAC

The southern Changjiang River Estuary has attracted considerable attention from marine scientists because it is a highly biologically active area and is biogeochemically significant. Moreover, land-ocean interactions strongly impact the estuary, and harmful algal blooms （HABs） frequently occur in the area. In October 2010 and May 2011, water samples of chromophoric dissolved organic matter （CDOM） were collected from the southern Changjiang River Estuary. Parallel factor analysis （PARAFAC） was used to assess the samples＇ CDOM composition using excitation-emission matrix （EEM） spectroscopy. Four components were identified： three were humic-like （C1, C2 and C3） and one was protein-like （C4）. Analysis based on spatial and seasonal distributions, as well as relationships with salinity, Chl a and apparent oxygen utilization （AOU）, revealed that terrestrial inputs had the most significant effect on the three humic-like Components C1, C2 and C3 in autumn. In spring, microbial processes and phytoplankton blooms were also important factors that impacted the three components. The protein-like Component C4 had autochthonous and allochthonous origins and likely represented a biologically labile component. CDOM in the southern Changjiang River Estuary was mostly affected by terrestrial inputs. Microbial processes and phytoplankton blooms were also important sources of CDOM, especially in spring. The fluorescence intensities of the four components were significantly higher in spring than in autumn. On average, C1, C2, C3, C4 and the total fluorescence intensity （TFI） in the surface, middle and bottom layers increased by 123%-242%, 105%-195%, 167%-665%, 483%-567% and 184%-245% in spring than in autumn, respectively. This finding corresponded with a Chl a concentration that was 16-20 times higher in spring than in autumn and an AOU that was two to four times lower in spring than in autumn. The humification index （HIX） was lower in spring that in autumn, and the fluorescence index （FI） was higher in spring than in autumn. This result indicated that the CDOM was labile and the biological activity was intense in spring.

Three-dimensional fluorescence characteristics of dissolved organic matter produced by Prorocentrum donghaiense Lu

Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrurn donghaiense at the exponential growth, stationary and decline stages into 〈0.45 μm filtrate, 100 kDa-0.45 μm, 1%100 kDa and 1-10 kDa retentate and 〈1 kDa ultrafiltrate fractions. The fluorescence properties of different molecular weights of dissolved organic matter （DOM） were measured by excitation-emission matrix spectra. Protein-like and humic-like fluorophores were observed in the DOM produced by P. donghaiense. The central positions of protein-like fluorophores showed a red shift with prolonged growth duration, shifting from tyrosine-like properties at the exponential growth stage to tryptophan-like properties at the stationary and decline stages. The excitation wavelengths of protein-like fluorophores exhibited some change in the exponential growth and stationary stages with increased molecular size, but showed little change in the decline stage. However, the emission wavelengths in the decline stage exhibited a blue shift. Very distinct C type and A type peaks in humic-like fluorophores were observed. With a prolonged culture time, the intensities of both of the peaks became strong and the excitation wavelengths of peak A showed a red shift, while the A：C ratios fell. More than 94% of fluorescent DOM was in the lower than 1 kDa molecular weight fraction.

Multivariate analysis of fluorescence and source identification of dissolved organic matter in Jiaozhou Bay, China

Hierarchical clustering analysis and principal component analysis （PCA） methods were used to assess the similarities and dissimilarities of the entire Excitation-emission matrix spectroscopy （EEMs） data sets of samples collected from Jiaozhou Bay, China. The results demonstrate that multivariate analysis facilitates the complex data treatment and spectral sorting processes, and also enhances the probability to reveal otherwise hidden information concerning the chemical characteristics of the dissolved organic matter （DOM）. The distribution of different water samples as revealed by multivariate results has been used to track the movement of DOM material in the study area, and the interpretation is supported by the results obtained from the numerical simulation model of substance tracing technique, which show that the substance discharged by Haibo River can be distributed in Jiaozhou Bay.

Quick Fluorescence Method for the Distinguishing of Vegetable Oils

This paper presents the possibilities offered by fluorescence spectroscopy for the identification of vegetable oils such as soybean, sunflower, flax, walnut, corn, almond, sesame, olive and pumpkin oils. The probes under study have been excited with two types of sources： a laser diode （LD） and light-emitting diodes （LEDs） emitting in the UV and in the visible range. Total luminescence spectra were recorded by measuring the emission spectra in the range 350-720 nm at excitation wavelengths from 375 to 450 nm. The excitation-emission matrices have been obtained and two basic fluorescence regions in the visible have been outlined. On this basis the fluorescence spectra of the oils have been subdivided into three categories depending on the prevalence of the fluorescence maxima. The samples show differences in their fluorescence spectra. The latter fact shows that fluorescence spectroscopy can be used for the quick identification of edible oils. The fatty acid, the tocopherol, the beta-carotene and chlorophyll contents in the analyzed oils have been studied. It is shown that some of the types of oils differ significantly from each other by the first derivatives of their fluorescence spectra. There also exist color differences between the groups of vegetable oils under study.

Excitation-emission matrix (EEM) fluorescence spectroscopy for characterization of organic matter in membrane bioreactors: Principles, methods and applications

The membrane bioreactor(MBR)technology is a rising star for wastewater treatment.The pollutant elimination and membrane fouling performances of MBRs are essentially related to the dissolved organic matter(DOM)in the system.Three-dimensional excitation-emission matrix(3D-EEM)fluorescence spectroscopy,a powerful tool for the rapid and sensitive characterization of DOM,has been extensively applied in MBR studies;however,only a limited portion of the EEM fingerprinting information was utilized.This paper revisits the principles and methods of fluorescence EEM,and reviews the recent progress in applying EEM to characterize DOM in MBR studies.We systematically introduced the information extracted from EEM by considering the fluorescence peak location/intensity,wavelength regional distribution,and spectral deconvolution(giving fluorescent component loadings/scores),and discussed how to use the information to interpret the chemical compositions,physiochemical properties,biological activities,membrane retention/fouling behaviors,and migration/transformation fates of DOM in MBR systems.In addition to conventional EEM indicators,novel fluorescent parameters are summarized for potential use,including quantum yield,Stokes shift,excited energy state,and fluorescence lifetime.The current limitations of EEM-based DOM characterization are also discussed,with possible measures proposed to improve applications in MBR monitoring.

Rapid determination of thiabendazole in orange extract using excitation-emission matrix fluorescence and second-order calibration based on alternating trilinear decomposition/alternating normalization-weighted error algorithms

A novel approach is proposed for direct quantitative analysis of thiabendazole in the orange extract by using excitation-emission matrix fluorescence coupled with second-order calibration methods based on the alternating trilinear decomposition(ATLD) and the alternating normalization-weighted error(ANWE) algorithms,respectively. The average recoveries of thiabendazole in the orange extract by using ATLD and ANWE with an estimated component number of two were 99.7 ± 3.3% and 103.5 ± 4.1%,respectively. Furthermore,the accuracy of the two algorithms was also evaluated through elliptical joint confidence region(EJCR) tests as well as figures of merit,such as sensitivity(SEN),selectivity(SEL) and limit of detection(LOD). The experimental results demonstrate that both algorithms have been satisfactorily applied to the determination of thiabendazole in orange extract,and the perform-ance of ANWE is slightly better than that of ATLD.

水源水Fe(Ⅵ)与Mn(Ⅶ)预氧化对天然有机物的组成特征及消毒副产物生成势的影响

溶解有机物(DOM)普遍存在于地表水源中,是饮用水消毒副产物的重要前驱物.本研究考察了Fe(Ⅵ)与Mn(Ⅶ)预氧化对浙江东部某水源水中DOM的组成特征及消毒副产物生成势的影响.运用三维荧光光谱(3D-EEMs)分析了两种氧化剂在不同的浓度梯度下对水样进行预氧化后,水样DOM的荧光光谱特性、荧光特征参数和荧光组分等的变化情况,及氯化后消毒副产物(DBPs)的生成情况.结果表明,水源水水样DOM的来源主要为陆源与内源混合且具有一定的腐殖质化程度,Mn(Ⅶ)和Fe(Ⅵ)预氧化不仅对荧光DOM具有较好的降解效果,对腐殖质类DOM也具有一定的降解作用.在DBPs的生成势方面,Mn(Ⅶ)预氧化对三氯甲烷(TCM)生成势的变化呈现出先增强后减弱的现象,对二氯乙腈(DCAN)的生成势具有一定的减弱作用.而Fe(Ⅵ)预氧化对三氯丙酮(1,1,1-TCP)、溴氯乙腈(BCAN)、二溴乙腈(DBAN)和DCAN的生成势均具有明显的降低作用,对二氯一溴甲烷(DCBM)、二溴一氯甲烷(DBCM)和三溴甲烷(TBM)生成势的变化呈现出先增强后减弱的现象.研究结果推动了水源水氧化消毒工艺的发展并为相关研究提供理论指导.

基于水质荧光指纹技术的复合污染河道溯源研究

复合污染水体的污染源组成解析是当前水污染溯源领域难题,基于三维荧光光谱的水质荧光指纹技术是解决该难题的关键方法。采用水质荧光指纹技术对苏南地区Y河开展溯源分析。通过平行因子分解与水质荧光指纹比对有效识别Y河中印染废水荧光信号来自上游河道异地输入。利用氨氮浓度划分重污染区域,依托水质荧光指纹对Y河流域内109个排口开展筛查,通过污染路径溯源法精准定位35家主要责任单位。落实整改要求后,Y河水质由劣Ⅴ类迅速提升至Ⅲ类,并持续稳定达标。Y河河道治理工程取消,降本增效效果显著。针对复合污染河道提出了以水质荧光指纹技术为抓手的精准溯源与微创治理新模式,对实现精准治污、科学治污、依法治污具有重要指导意义。

FCC催化剂传质强化活性位利用效率的可视化分析

流化催化裂化(FCC)催化剂传质性能抑制活性位利用效率是影响其催化性能的关键因素,选用6种不同基质材料制备的FCC催化剂样品,在系统分析催化孔结构和酸性质的基础上,利用激光共聚焦荧光显微成像技术和吸附穿透曲线法模拟考察了重油大分子的传质扩散行为;并运用单分子荧光成像技术,以B酸中心催化噻吩低聚反应生成低聚物的数量作为判据考察了系列催化剂的活性位利用效率。研究发现,相比于传统的高岭土和拟薄水铝石作为基质材料,本课题组设计开发的大孔富B酸的基质材料(APM-9)明显优化了大分子在FCC催化剂上的传质性能,从而显著提升了活性位的利用效率。发展了一种新颖的FCC催化剂结构参数与其传质性能、酸中心利用效率构效关系建立的方法,可为催化剂基质材料的设计和优化提供数据支撑和理论指导。

C16醇改性SiO 2基体材料对荧光强度的影响因素研究

黄磷炉渣大量堆积不仅会造成资源浪费,还会污染环境,针对黄磷炉渣能够得到合理利用,研究了以黄磷炉渣为主要原料,经过硝酸酸浸制备SiO_(2)这一工艺流程。因SiO_(2)生物相容性好、化学稳定性高、表面易于功能化等优点,可以作为制备荧光材料的基体材料。稀土金属Ce^(3+)直接掺杂到SiO_(2)上很难实现,所以SiO_(2)的表面改性是稀土金属掺杂SiO_(2)基制备荧光材料的一项关键因素,C16醇原料便宜,操作简单,并且改性效果明显,能够显著提高SiO_(2)基的接枝率,因此采用C16醇对SiO_(2)进行表面改性。为了探究优化的实验条件,考察C16醇投入量、改性时间、溶剂体积、改性温度和洗水用量对材料荧光强度的影响。结果表明:0.2g C16醇与30mL乙醇溶剂在175℃温度下反应2h,洗水用量为70mL为优化改性条件。

高盐食品基质中砷含量的测定

建立了一种基体匹配-动态反应池-电感耦合等离子体质谱法测定高盐食品基质中砷含量的方法,以解决高盐基体对砷测定的质谱干扰和基体干扰。研究电感耦合等离子体质谱法与氢化物发生原子荧光光谱法测定食品中总砷的技术要点,通过分析前处理消解方式、有机基质、氯含量和内标校正元素等对砷含量测定的影响,建立了高盐食品基质中砷含量的测定方法。实验结果表明,微波消解法并不太适用于氢化物发生原子荧光光谱法测定食品中总砷,有机基质和氯分别会对砷的质谱测定信号产生增敏效应和质谱干扰,74Ge比72Ge更适合作为电感耦合等离子体质谱法测定食品中总砷的内标元素。建立的检测方法定量限0.010 mg/kg,高盐基体中砷在0.010、0.025、0.20、0.50 mg/kg添加水平的回收率为91.50%~110.80%,相对标准偏差范围为0.76%~7.22%。本法和参照法对样品分析结果统计值均大于t0.90,10(1.81,双边),测定结果存在显著性差异,对实际样品测定结果表明动能歧视测定模式并不太适用于高盐食品基质样品中总砷的测定。该方法准确可靠,能用于高盐和高基体食品中砷含量的测定分析。

塑化剂对聚合物基质荧光猝灭型氧敏感膜性能影响的研究

本研究以四(五氟苯基)卟吩铂(PtTFPP)为荧光指示剂,聚氯乙烯(PVC)为基质,柠檬酸三丁酯(TBC)为塑化剂,制备荧光猝灭型氧敏感膜。通过刮刀涂膜工艺制备样品,并利用自建的基于相位调制的荧光寿命测量装置,测试氧敏感膜的荧光猝灭性能。研究发现,添加质量比40%的TBC显著提高了PVC的氧气渗透速率,增强了荧光猝灭效应,导致在20kPa氧分压下荧光寿命从69.8μs大幅度降至16.3μs,反映灵敏度大小的KSV常数从0.013kPa^(-1)增大到0.375kPa^(-1)。对于添加TBC的氧敏感膜,其荧光寿命随氧分压的变化符合双位点Stern-Volmer模型,究其原因是膜表层和内部的氧浓度存在差异,导致荧光猝灭效应呈现较强和较弱两部分。当气氛由氮气切换到空气时,其荧光相位信号对气相氧分压变化的响应时间为4.5s,远小于不含TBC的样品30.5s。本研究的结果有助于改进以聚合物为基质的荧光猝灭型氧传感器的性能。

Toward a rapid and convenient nanoplastic quantification method in laboratory-scale study based on fluorescence intensity

The thorough investigation of nanoplastics(NPs)in aqueous environments requires efficient and expeditious quantitative analytical methods that are sensitive to environmentally relevant NP concentrations and convenient to employ.Optical analysis-based quantitative methods have been acknowledged as effective and rapid approaches for quantifying NP concentrations in laboratory-scale studies.Herein,we compared three commonly used optical response indicators,namely fluorescence intensity(FI),ultraviolet absorbance,and turbidity,to assess their performance in quantifying NPs.Furthermore,orthogonal experiments were conducted to evaluate the influence of various water quality parameters on the preferred indicator-based quantification method.The results revealed that FI exhibits the highest correlation coefficient(>0.99)with NP concentration.Notably,the limit of quantification(LOQ)for various types of NPs is exceptionally low,ranging from 0.0089 to 0.0584 mg/L in ultrapure water,well below environmentally relevant concentrations.Despite variations in water quality parameters such as pH,salinity,suspended solids(SS),and humic acid,a robust relationship between detectable FI and NP concentration was identified.However,an increased matrix,especially SS in water samples,results in an enhanced LOQ for NPs.Nevertheless,the quantitative method remains applicable in real water bodies,especially in drinking water,with NP LOQ as low as 0.0157–0.0711 mg/L.This exceeds the previously reported detectable concentration for 100 nm NPs at 40µg/mL using surface-enhanced Raman spectroscopy.This study confirms the potential of FI as a reliable indicator for the rapid quantification of NPs in aqueous environments,offering substantial advantages in terms of both convenience and cost-effectiveness.

尿液基细胞荧光原位杂交检测对膀胱尿路上皮癌的诊断价值

目的探讨尿液基细胞学(LBC)靶向的荧光原位杂交(FISH)对膀胱尿路上皮癌(BUC)的诊断价值。方法回顾性收集2020年10月—2022年10月于首都医科大学附属北京天坛医院泌尿外科行膀胱镜检查的128例患者的临床资料。所有患者在行膀胱镜检查前进行尿核基质蛋白22(NMP22)检测、尿LBC检测与尿LBC靶向的FISH检测,以术后病理结果为标准,分析3种检查方法的灵敏度和特异度。结果NMP22、尿LBC与LBC靶向的FISH的灵敏度分别为61.11%、79.17%、82.46%,特异度分别为57.14%、73.21%、86.67%;NMP22、尿LBC的灵敏度在检测高级别BUC时优于低级别,差异有统计学意义(P=0.01,P=0.03);3种方法对肌层浸润性膀胱癌与非肌层浸润性膀胱癌检测的灵敏度比较,差异无统计学意义(P≥0.05)。结论尿LBC靶向的FISH对BUC诊断的灵敏度和特异度较高,尤其是对于低级别BUC,可以作为BUC早期筛查、诊断的重要方法。

Impact of transparent exopolymer particles on the dynamics of dissolved organic carbon in the Amundsen Sea,Antarctica

The Southern Ocean is an important carbon sink pool and plays a critical role in the global carbon cycling.The Amundsen Sea was reported to be highly productive in inshore area in the Southern Ocean.In order to investigate the influence of transparent exopolymer particles(TEP)on the behavior of dissolved organic carbon(DOC)in this region,a comprehensive study was conducted,encompassing both open water areas and highly productive polynyas.It was found that microbial heterotrophic metabolism is the primary process responsible for the production of humic-like fluorescent components in the open ocean.The relationship between apparent oxygen utilization and the two humic-like components can be accurately described by a power-law function,with a conversion rate consistent with that observed globally.The presence of TEP was found to have little impact on this process.Additionally,the study revealed the accumulation of DOC at the sea surface in the Amundsen Sea Polynya,suggesting that TEP may play a critical role in this phenomenon.These findings contribute to a deeper understanding of the dynamics and surface accumulation of DOC in the Amundsen Sea Polynya,and provide valuable insights into the carbon cycle in this region.