[Rapid Quantitative Detection of Bacterial in Water Based on Multi-Wavelength Scattering Spectra].

The structures of bacterial cells are analyzed in this paper. The scattering components of individual cell were divided into two parts including external structure and internal structure. The interpretation model of bacteria about scattering light is established. The model is used to analyze the scattering light of Escherichia coli in the region of 400~900 nm. The average size of external structure and the internal structure can be obtained, and the ratio of the two parts is also obtained. According to the relationship of the optical density of single cell and the overall measurement, the concentration of bacterial can be obtained quickly. The maximum difference in all the concentrations of the bacteria repeated measurements is 1.83%; compared with the plate culture method, the measurement results were in the same order of magnitude, with relative error of 3.43%. The scattering light of Escherichia coli and Klebsiella pneumoniae are analyzed in different growth stages, the curves of the concentration and the size of the two species bacteria over time are obtained. The results can provide a quick way for the study of bacterial growth and technical support for rapid detection of bacteria in the water.

[Study of the Impact of Sample Thickness on Thin Film Method X-Ray Fluorescence Spectrum Measurement].

The mixed samples of nylon film enrichment of Cr, Pb and Cd three elements and glass fiber membrane filter were as the research object. With the method of superposition of membrane filter, the XRF spectra were measured under different thin film samples thicknesses. According the changes of characteristic XRF of Cr, Pb and Cd elements in the mixed sample and Ca, As and Sr elements in glass fiber membranes, the effects of sample thickness on thin film method XRF spectrum measurement were studied. The study results showed that the effects of thin film sample thickness on the fluorescent properties of elements with characteristic spectral lines in different energy ranges were different. The energy of characteristic spectral lines was greater, the loss of element characteristic X-ray fluorescence when it passed through membrane and reached detector was less. But matrix effect caused by thin film sample thickness increase was stronger with the energy of characteristic spectral lines greater. The background fluorescent intensity in corresponding characteristic spectral line location was greater. So the impact of matrix effect caused by sample thickness increase on thin film method XRF spectrum measurement sensitivity was greater. For elements with low energy characteristic spectral lines (energy≤7 keV), the way of increasing thin film sample thickness in order to increase the mass-thickness concentration of component measured, can not effectively improve the sensitivity of thin film method XRF spectrum measurement. And thin film samples thickness within 0.96 mm is conductive to the measurement and analysis of XRF spectrum. For element with higher energy characteristic spectra lines(energy＞7 keV), the sensitivity of XRF spectrum measurement can be appropriately increased by the way of increase thin film sample thickness in order to increase the mass-thickness concentration of component measured. And thin film samples thickness within 0.96～2.24 mm is more conductive to the measurement and analysis of XRF spectrum. The study provides an important theoretical basis for thin sample preparation and enrichment technology of thin film method X-ray fluorescence spectrum analysis the atmosphere and water heavy metal.

[Effect of Methanol-Water Mixture on Three-Dimensional Fluorescence Spectra of Carbaryl].

Extensive use of pesticides has a significant impact on the environment. Carbaryl, whose residues stay in the surface water, is one of the most widely used broad spectrum insecticides in agriculture. It is important to understand carbaryl spectral characteristics and detection methods. The characteristic of excitation-emission three-dimensional spectra of carbaryl is studied. By changing the concentration of methanol in methanol-water binary solvent, the impact of methanol-water mixture on three-dimensional fluorescence spectra of carbaryl is discussed. The results show that the characteristic excitation-emission spectra of carbaryl is single peak, the range of the excitation wavelength and emission wavelength are: 244~304 and 300~350 nm respectively, the maximum excitation/emission peak located at 280 and 335 nm. With increasing the content of methanol in methanol-water binary solvent mixture, there is no obviously spectra shift of three dimensional fluorescence spectra of carbaryl. However, the intensity of fluorescence is nonlinear dependent on the content of methanol, mainly due to the specific properties of binary mixed solvent.

[The study of selecting sample detecting position and lead plate inner material in thin film method X-ray fluorescence measurement].

(1) In this paper type 316 stainless steel metal plate as the research object, the selection of sample detecting position was studied when thin film method X-ray fluorescence measurement was conducted. The study showed that the optimal location for the sample detection was sample distance X-ray tube and detector baseline 1cm with the baseline into a 16°angle. (2) Heavy metal pollutants of Pb, Cd and Cr in industrial ambient air as the main analysis object, when thin film method X-ray fluorescence conducted with lead plate protection, X-rays will penetrate the membrane and continuely stimulate the protective lead plate. Therefore there is lead spectral line interference in the filter membrane background spectrum, which will affect the detection of lead element in real samples. Studies show that when a layer of isolating material was applied between the thin sample and the protective lead plate, the interference of lead line can effectively be avoided. (3) Several rigid insulating material of type 316 stainless steel, brass, aluminum, red copper and PTEE as lead inner material were selected and studied. The study results showed that compared with X-ray fluorescence spectra of other lead inner materials, the X-ray fluorescence spectrum of red copper contained the least element spectral lines. There were not Cr, Cd and Pb spectrum peaks in the X-ray fluorescence spectrum of red copper. And the target timber scattering spectrum intensity in the high energy part was weaker compared to other X-ray fluorescence spectrum. The above analysis shows that red copper has the minimal disturbance to the actual measurement of heavy metals Cr, Cd and Pb. At the same time, red copper as lead inner materials can effectively avoid the interference of lead spectrum line in lead plate. So red copper is the best lead plate inner materials in thin film method X-ray fluorescence spectroscopy measurement. This study provides an important theoretical basis for the assembling and setting'up air and water weight metal X-ray fluorescence spectrometer.

[Photosynthetic Parameters Inversion Algorithm Study Based on Chlorophyll Fluorescence Induction Kinetics Curve].

The fast chlorophyll fluorescence induction curve contains rich information of photosynthesis. It can reflect various information of vegetation, such as, the survival status, the pathological condition and the physiology trends under the stress state. Through the acquisition of algae fluorescence and induced optical signal, the fast phase of chlorophyll fluorescence kinetics curve was fitted. Based on least square fitting method, we introduced adaptive minimum error approaching method for fast multivariate nonlinear regression fitting toward chlorophyll fluorescence kinetics curve. We realized Fo (fixedfluorescent), Fm (maximum fluorescence yield), σPSII (PSII functional absorption cross section) details parameters inversion and the photosynthetic parameters inversion of Chlorella pyrenoidosa. And we also studied physiological variation of Chlorella pyrenoidosa under the stress of Cu(2+).

[Determination of chemical rank of three-dimensional fluorescence spectra using mathematical morphology method].

The present paper firstly denoises the signal with morphological method, selecting sine-shaped structure element, using the morphological difference in waveform between the three-dimensional fluorescence and noise signal, then singular value decomposition is applied to the denoised data, and finally the chemical rank is determinated jointing eigenvalues and eigenvectors form singular value decomposition. This paper principally discusses the theory basis of morphological filtering method, firstly simulated data is analysed by morphological filtering method to confirm the necessity and effectiveness of proposed method, then the feasibility and practicability of the proposed method is verified by the determination of components number of phenols mixture three-dimensional fluorescence spectra compared with traditional Monte Carlo method. The experiments demonstrate that the proposed method is able to estimate the chemical rank correctly.

[Analysis of three-dimensional fluorescence overlapping spectra using nonnegative matrix factorization].

The present paper primarily tests and verifies the effect of NMF in blind source separation of three-dimensional simulative fluorescence spectra, and then four different computational algorithms (multiplicative iterative; alternating least square; second order method; projected gradient algorithm) were used in three practical phenolic compounds (cresol, phenol, thymol) overlapping fluorescence spectra to find out which nonnegatively constrained algorithms is the most efficient for fluorescence spectra unmixing. The experiments demonstrate that four ways have the normalized residuals below 0.06%, and alternating least square (ALS) is the best at both convergence behavior and robustness.

[Analysis of three-dimensional fluorescence overlapping spectra using differential spectra and independent component analysis].

The analysis of multi-component three-dimensional fluorescence overlapping spectra is always very difficult. In view of the advantage of differential spectra and based on the calculation principle of two-dimensional differential spectra, the three-dimensional fluorescence spectra with both excitation and emission spectra is fully utilized. Firstly, the excitation differential spectra and emission differential spectra are respectively computed after unfolding the three-dimensional fluorescence spectra. Then the excitation differential spectra and emission differential spectra of the single component are obtained by analyzing the multicomponent differential spectra using independent component analysis. In this process, the use of cubic spline increases the data points of excitation spectra, and the roughness penalty smoothing reduces the noise of emission spectra which is beneficial for the computation of differential spectra. The similarity indices between the standard spectra and recovered spectra show that independent component analysis based on differential spectra is more suitable for the component recognition of three-dimensional fluorescence overlapping spectra.

[Studies on toxicity of four kinds of heavy metals in water by synchronous-scan fluorescence].

Spectrofluorometry of chlorella pyrenoidosa was studied by three dimensional excitation-emission (3DEEM) fluorescence spectroscopy and synchronous scan fluorescence spectroscopy with Delta gamma = 20 nm in the stress of Hg+, Cd2+, Cu2+ and Zn2+. The conclusion from two kinds of Spectrofluorometry was the same: after 96h stress by heavy metals, the maximum fluorescence values reduced obviously, chlorophyll-a and chlorophyll-b in the photosynthetic system were seriously damaged by heavy metal. Further analysis of the correlations between heavy metal concentration and fluorescence quenching efficiency I0/I can conclude that the toxicity of heavy metal and the fluorescence quenching efficiency I0/I were positively correlated, and they all increased with the heavy metal concentration and stress time. For one kind of heavy metals, synchronous scan fluorometry is a sensitive method for its toxicity assessment. Compared to 3DEEM fluorescence spectroscopy, synchronous scan fluorescence spectroscopy is less time consuming and of higher selectivity. It is suitable to assess the toxicity of pollutions in water.

[The method of phytoplankton photosynthesis activity in-situ measurement based on light induced fluorescence].

According to the phytoplankton fluorescence induction characteristics under different light conditions, chlorophyll fluorescence as a probe for analysis of phytoplankton photosynthesis was studied. The present paper proposed a in-situ measurement method based on the chlorophyll fluorescence values Ft and Fm to get phytoplankton photosynthesis activity, Chlorella vulgaris, microcystis aeruginosa and Cyclotella meneghiniana Kiits were selected as experimental subjects, a comparison test was done between self-developed in-situ measurement system and Water PAM in lab, and the results showed that coefficients between the two methods were 0.9778, 0.8786 and 0.7953. This work provides a rapid and in-situ measurement method for phytoplankton photosynthesis activity.

[Experimental studies on three-dimensional fluorescence spectral of mineral oil in ethanol].

Three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy was applied to characterize the mineral oil in ethanol. In the paper, a simple way of treating the first-order Rayleigh, Raman and second-order Rayleigh scatter is used involving subtracting an EEM of a solvent blank, and removing and replacing the values with zeros. The corrected results showed that the characteristic three-dimensional fluorescence spectra of mineral oil were as follows: there was one mainly peak in the fluorescence spectrum of the kerosene, and the peak was identified at excitation/emission wavelengths (Ex/Em) of 270/290 nm; there were two intense fluorescence peaks in the fluorescence spectrum of 0 # diesel, located at (Ex/Em) of 240/344 nm and 270/362 nm respectively; there were several fluorescence peaks in the fluorescence spectrum of lubricating oil, with the two intense peaks located at (Ex/Em) of 240/348 nm and 258/358 nm respectively. Furthermore, the relation between the relative fluorescence intensity and the concentration of mineral oil was studied, and the sensitivity and the limitation of detection of the method were also analyzed. The results of this study show that the method of three-dimensional fluorescence can be used to detect the low concentration of oil.

[Excitation-emission fluorescence characterization study of the three phenolic compounds].

The fluorescence intensity information was collected by scanning its fluorescence spectra at different excitation wavelengths. Based on its high sensitivity and selectivity, excitation-emission fluorescence can be widely used for detection of pollutants in the environment. The characterizations of the three phenolic compounds were investigated by this method, and the fluorescence peaks of phenol, m-cresol and thymol were confirmed at 272/300 nm, 274/300 nm and 276/304 nm when the excitation and emission wavelength were set in the range of 240-360 nm and 260-500 nm respectively. The excitation and emission spectra of the three phenolic compounds are very similar because of their analogical structure. The intensity of the spectrum has a good linear relationship with the concentration when the solution concentration is between 0.02 and 1.0 mg x L(-1), and the limits of detection can reach at 1 microg x L(-1). Results show that the three compounds can be analyzed qualitatively and quantitatively by excitation-emission fluorescence.

Simultaneous measurement of phenols by three-way fluorescence spectroscopy: A comparison of N-PLS/RBL, U-PLS/RBL and PARAFAC.

Phenol, o-cresol, p-cresol, catechol and resorcinol are five phenolic compounds with extremely similar structure. Their fluorescence spectra are hard to be analyzed because of the serious spectral overlaps between any two of the five phenolic components in the mixture system. In this experiment, multi-dimensional partial least-squares (N-PLS), unfolded partial least-squares (U-PLS) with residual bilinearization (RBL) and parallel factor analysis (PARAFAC) are employed to analyze the three-way fluorescence spectra aiming to achieve quantitative results. Meanwhile, a contrast of these three methods is given. The experiment results show that N-PLS/RBL and U-PLS/RBL algorithms are superior to PARARFAC in terms of analysis of highly overlapping three-way fluorescence spectra for concentration determination.

[Quantitative analysis of phenanthrene and humic acid in solution based on fluorescence method].

Polycyclic aromatic hydrocarbons as a universal concern in the monitoring of priority pollutants have low content in the water environment and are interfered with the coexistence of humic acid. The spectra of humic acid and polycyclic aromatic hydrocarbons overlap seriously, so it was difficult to use the conventional methods for rapid quantitative detection. As a model compound of polycyclic aromatic hydrocarbons, Phenanthrene (PHE) was chosen in the experiment. The fluorescence spectra of PHE and humic acid (HA) were investigated by three dimensional fluorescence excitation-emission matrix. The method combined excitation-emission matrix fluorescence spectra with parallel factor analysis and was applied to determine PHE and HA directly. The excitation wavelength changed from 240 to 360 nm at an interval of 5 nm. The emission wavelength varied from 260 to 575 nm at an interval of 5 nm. The satisfactory results show that this experiment can be easily performed without paying out time-consuming and complicated procedures.

[Study on three-dimensional fluorescence spectra of phenanthrene].

According to the high fluorescence quantum yields of polycyclic aromatic hydrocarbons (PAHs), the fluorescence spectra of phenanthrene were investigated by three dimensional fluorescence excitation-emission matrix (3DEEM). The results show that the three-dimensional fluorescence spectra of phenanthrene in aqueous solution mainly have two fluorescence peaks. On the basis of three-dimensional fluorescence spectrometry analysis of phenanthrene, the excitation wavelength of 255 nm and emission wavelength of 273 nm were chosen for the quantitative analysis of phenanthrene. The linear range for the determination of phenanthrene was 5.0-250.0 mg x mL(-1), its detection limit was 3. 88 ng x mL(-1), and its relative standard deviation was 4.23% (n=5). It was a good precision. It has been tested satisfactorily for the determination of artificial sample in tap water. The recoveries are in the range of 90%-105%. The method provided basis for the rapid monitoring of trace PAHs in water.

[Study on a new method of fast monitoring toxicity of Cd2+ by algal in water].

Chlorophyll concentration and photosynthesis activity fluorescence parameters of Chlorella pyrenoidosa stressed by different concentrations of Cd2+ were measured based on algal growth inhibition tests and photosynthetic activity inhibition tests. The relationship between the algal photosynthetic activity inhibition rate and 96 h inhibition rate of specific growth rate at different Cd2+ stress times was studied by sigmoidal curve fitting and one-way ANOVA analysis. The result shows that S function relevance exists between the algal photosynthetic activity inhibition rates for 48 h, 53 h, 72 h, 77 h and 96 h respectively and 96 h inhibition rate of specific growth rate (R2 > 0.95). Consequently, EC10 (10% effective concentration) after 48 h and 53 h inhibition in photosynthetic activity inhibition tests could be used to represented EC50 (50% effective concentration) in 96 h algal growth inhibition tests for evaluating the Cd2+ toxicity. Dose-response relationships between the algal photosynthetic activity inhibition rates after 48 h and 53 h inhibition and Cd2+ toxic equivalency quantity were further analyzed. The method provided a rapid and viable new thought to monitoring single Cd2+ toxicity in lab and early warn integrated toxicity of pollution in water.

Hydrothermal synthetic mercaptopropionic acid stabled CdTe quantum dots as fluorescent probes for detection of Ag⁺.

Mercaptopropionic acid (MPA) capped CdTe quantum dots (QDs) with particle size 3 nm have been successfully synthesized in aqueous medium by hydrothermal synthesis method. And the effects of different metal ions on MPA capped CdTe QDs fluorescence were studied using fluorescence spectrometry. The results demonstrated that at the same concentration level, Ag(+) could strongly quench CdTe QDs fluorescence, and the other metal ions had little effect on CdTe QDs fluorescence except Cu(2+). On the basis of this fact, a rapid, simple, highly sensitive and selective method based on fluorescence quenching principle for Ag(+) detection in aqueous solution was proposed. Under optimal conditions, the quenched fluorescence intensity (F(0)-F) increased linearly with the concentration of Ag(+) ranging from 4 × 10(-7) to 32 × 10(-7)mol L(-1). The limit of detection for Ag(+) was 4.106 × 10(-8)mol L(-1). The obtained plot of F(0)/F versus [Ag(+)] was an upward curvature, concave towards the y-axis, rather than a straight line. The modified form of the Stern-Volmer equation was third order in Ag(+) concentration. According to the modified Stern-Volmer equation, it can be inferred that dynamic quenching and static quenching simultaneously occurred when Ag(+) interacted with MPA capped CdTe QDs. At the same time other factors might also influence the quenching process. Based on this study, hydrothermal synthesized MPA capped CdTe QDs with particle size 3 nm may be used as a novel fluorescence probe to quantificationally and selectively detect Ag(+).