Optical properties and nondestructive estimation of anthocyanin content in plant leaves.

Absorption and reflectance spectra of maple (Acer platanoides), cotoneaster (Cotoneaster alaunica), dogwood (Cornus alba) and pelargonium (Pelargonium zonale) leaves with a wide range of pigment content and composition were studied in visible and near-infrared spectra in order to reveal specific anthocyanin (Anth) spectral features in leaves. Comparing absorption spectra of Anth-containing and Anth-free leaves with the same chlorophyll (Chl) content, absorption spectra of Anth in leaves were derived. The main spectral feature of Anth absorption in vivo was a peak around 550 nm; the peak magnitude was closely related to Anth content. A quantitative nondestructive technique was developed to subtract Chl contribution to reflectance in this spectral region and retrieve Anth content from reflectance over a wide range of pigment content and composition. Anth reflectance index in the form ARI = (R550)-1 - (R700)-1, where (R550)-1 and (R700)-1 are inverse reflectances at 550 and 700 nm, respectively, allowed an accurate estimation of Anth accumulation, even in minute amounts, in intact senescing and stressed leaves.     

Diffuse reflectance spectroscopy of powders

Diffuse reflectance (DRIFT) spectra in the mid-IR region, especially of inorganic powders, often reveal inverted or derivative-like structures (“reststrahlen” bands) which make their straightforward interpretation in terms of the Kubelka-Munk theory difficult. A simulation technique based on a combination of single scattering (Mie theory) and radiative transfer (with Kubelka-Munk as a special case) is reported that allows measured DRIFT spectra to be described without use of any free “fit” parameters. The results for silica glass microspheres and silicon carbide powder show that not specular reflectance but prominent structures in the single particle scattering cross-section are responsible for the reststrahlen features which cannot be suppressed even by diluting the powder in an excess of KBr. The efficiency of a “blocker” device in DRIFTS is also discussed.     

Diffuse reflectance Fourier transform IR spectroscopy

Fourier transform infrared (FTIR) spectroscopy has greatly enhanced the utility and range of applications of infrared (IR) spectroscopy in chemistry. Until the development of routine FTIR instrumentation, applications of IR were essentially limited to direct transmission measurements. Newer sampling methods in FTIR spectroscopy can be successful with less than 1% throughput of the infrared radiation and yet yield a spectrum with acceptable signal-to-noise ratio. The decreasing cost of FTIR spectrometers has made a number of such sampling techniques accessible for routine applications without compromises in performance. This article will describe one of these, diffuse reflectance.     

Diffuse reflectance spectroscopy—Important developments

Effect of front surface reflectance is incorporated into standard Kubelka-Munk theory of diffuse reflection. A method of obtaining the absorption coefficient independent of scattering coefficient is presented.     

Use of near-infrared reflectance spectroscopy in predicting nitrogen,phosphorus and calcium contents in heterogeneous woody plant species

Near-infrared reflectance spectroscopy was applied to determine nitrogen (N), phosphorus (P) and calcium (Ca) content in leaf samples of 18 woody species. A total of 183 samples from mountain, riparian and dry areas from the Central–Western Iberian Peninsula were collected for this purpose. The wide intervals of variation observed in nutrient concentrations (6.6–45.0 g kg^–1 for N, 0.24–2.97 g kg^–1 for P, and 1.00–20.06 g kg^–1 for Ca) were due to the great heterogeneity of the samples. To develop calibration equations, multiple linear regression, and partial least-squares regression (PLSR) were used. In both cases, three mathematical transformations of the data were applied: log1/R and first and second derivatives. The best calibration statistics were obtained using PLSR and derivative transformations (second derivative for N and first derivative for P and Ca). The following coefficients of multiple determination (R²) and standard errors of cross validation were obtained: 0.99 and 0.93 for N, 0.94 and 0.15 for P, and 0.95 and 0.88 for Ca. In the external validation the standard errors of prediction obtained were 0.76 (N), 0.11 (P) and 0.60 (Ca).     

近红外光谱法分析土壤中的有机质和氮素

应用近红外光谱技术测定土壤中的全氮、有机质、碱解氮，分别测定了2mm、0.15mm粒度的风干土在4000cm^-1～12000cm^-1波数范围的近红外光谱，用偏最小二乘法建立数学模型来进行含量预测，结果表明近红外光谱与土壤有机质、全氮、碱解氮具有良好的相关性，2mm风干土碱解氮建模的决定系数R^2为92.39，相对标准偏差为7.5%；2mm风干土全氮建模的决定系数R^2为88，相对标准偏差为8.2%;0.15mm的全氮建模的决定系数R2为89.86，相对标准偏差为7.2%;0.15mm风干土有机质建模的决定系数R^2为96.41，相对标准偏差为8.3%。因此，用近红外光普法测定土壤有机质、全氮、碱解氮的含量是可行的。     

Dynamic examinations at photoresists by reflectance spectroscopy

Summary An optical device is introduced, which allows the observation of the changes of chemical and physical properties of photoresists during irradiation. The photoresists are fixed as thin films at the surface of either semiconductor elements or circuit boards. The apparatus allows on-line process control during the photochemical reactions taking place in photoresists. By use of fiber optics and a photodiode array the spectral reflectance can be measured repetitively within 15 ms. This fast repetition rate allows on-line process control even during short radiation times, which are usual in practice to produce a pattern in the photoresists for the micro structuring of circuit boards or semiconductor wafers. The possibilities of the method and details of the apparatus are demonstrated at given examples of photoresists.

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Dynamische Untersuchungen an Photoresisten über Reflexionsspektroskopie

     

Internal reflectance spectroscopy associated with cyclic staircase voltammetry using a microcomputer

In order to obtain extensive information on electrode-solution interfaces, a rapid-scan internal reflectance spectroscopic system is combined with cyclic staircase voltammetry; the staircase wave generated by computer software is applied to an optically transparent tin oxide electrode. The internal reflectance spectrum is measured at each step of the applied staircase voltage simultaneously with the instantaneous current.     

Optimization of sample packing in diffuse reflectance infrared spectroscopy

Parameters of sample packing in diffuse reflectance IR Fourier transform spectroscopy (DRIFTS) have been investigated and optimized. One MPa pressure applied for 1 min was found to provide simultaneously at least ±3% reproducibility and appropriate band intensity. It has been also demonstrated that this precision can be obtained if the sample mass is controlled to ±2% and the sample is dry. A simple sample packing accessory was designed and constructed for reproducible sample preparation. Quartz concentration of dust samples has been determined and good agreement has been found with the results of the conventional pellet technique.     

Application of reflectance micro-infrared spectroscopy in coal structure studies

Fourier transform infrared micro-scale analysis of organic matter has been applied to characterise individual petrographic components of coal. The use of an infrared microscope made it possible to study and compare different parts of a single individual grain of vitrinite. This method enables analysis of particular macerals without the need for previous separation from coals. For this purpose the polished grainy micro-sections of coal have been prepared. The results obtained indicated the progression of aromatisation in the vitrinite structure with an increase of coalification degree of the parent coals.     

Determination of copper in urine by diffuse reflectance spectroscopy

A possibility of determining copper(II) after adsorption on polyacrylonitrile fiber filled with a KU-2 cation exchanger with immobilized 1-(2-pyridylazo)-2-naphthol (PANF–KU-2–PAN) is studied. The diffuse reflectance spectra of the carrier discs before and after the adsorption of copper in the batch mode are recorded. The dependences of the analytical signals of the copper complex on PANF–KU-2–PAN on adsorption conditions are studied. The conditions for determining 0.05–0.40 μg/mL of copper(II) by measuring diffuse reflection coefficient at 640 nm or visually by a color scale after the adsorption of copper from 20 mL of a solution containing 0.01 M of HCl are found. The determination of 0.1 μg/mL of copper does not interfered with equal amounts of Co, Zn, and Pb and double amounts of Ag, Fe(III), Cd, Mn(II), Bi(III), and Cr(III). The procedure is used for urine analysis. The following conditions of the preparation of urine samples for the determination of copper were selected: boiling with hydrogen peroxide followed by acidification to pH 2. A procedure for determining copper in urine with a limit of detection of 0.03 μg/mL was developed; the relative standard deviation does not exceed 25%; the duration of analysis of 5–6 samples is 30–35 min.     

Direct determination of lycopene content in tomatoes (Lycopersicon esculentum) by attenuated total reflectance infrared spectroscopy and multivariate analysis

Lycopene is a potent antioxidant that has been shown to play critical roles in disease prevention. Efficient assays for detection and quantification of lycopene are desirable as alternatives to time- and labor-intensive methods. Attenuated total reflectance infrared (ATR-IR) spectroscopy was used for quantification of lycopene in tomato varieties. Calibration models were developed by partial least-squares regression (PLSR) using quantitative measures of lycopene concentration from liquid chromatography as reference method. IR spectra showed a distinct marker band at 957 cm(-1) for trans Carbon-Hydrogen (CH) deformation vibration of lycopene. PLSR models predicted the lycopene content accurately and reproducibly with a correlation coefficient (sigma) of 0.96 and standard error of cross-validation <0.80 mg/100 g. ATR-IR spectroscopy allowed for rapid, simple, and accurate determination of lycopene in tomatoes with minimal sample preparation. Results suggest that the ATR-IR method is applicable for high-throughput quantitative analysis and screening for lycopene in tomatoes.     

Monitoring PDT by means of superficial reflectance spectroscopy

Monitoring of relevant parameters during photodynamic therapy (PDT) and correlating these with treatment response is necessary to guarantee optimal and reproducible treatment outcome. In this paper we study the correlation between changes in the local tissue optical properties (absorption and scattering coefficients) during ALA-PDT and changes in PpIX fluorescence. The optical properties are measured extremely superficially by employing a single fiber for the delivery and collection of white light to and from the tissue. The measured reflectance spectrum is modeled in terms of four relevant parameters: blood saturation, relative blood volume fraction, scattering intensity and wavelength dependence of the scattering. All these parameters, except the relative blood volume fraction, are shown to correlate with the rate of photobleaching of PpIX, which in turn has previously been shown to correlate with the response of tissues to PDT. These results yield valuable insight in the behavior of these parameters during PDT and their suitability to predict PDT-response for other photosensitizers for which monitoring through photobleaching is not possible.     

Metathesis polymerization-derived monolithic membranes for solid-phase extraction coupled with diffuse reflectance spectroscopy

Novel monolithic disks were prepared via ring opening metathesis polymerization (ROMP) from norborn-2-ene (NBE), a crosslinker, i.e., 1,4,5,8,8a-hexahydro-1,4,4,5,8, exo, endo-dimethanonaphthalene (DMN-H6) and tris(norborn-5-ene-2-ylmethylenoxy)methylsilane, respectively, 2-propanol and toluene (25:25:41:9, all %, w/w) using RuCl2(PCy3)2(CHPh) (Cy=cyclohexyl) as initiator and triphenylphosphine (PPh3) as modulator. Disks 1-2 mm thick were prepared by polymerization in a mold, disks thinner than 1mm were prepared by impregnation of nylon or other porous filters prior to the polymerization step. These disks were evaluated for the preconcentration of iodine and selected organic solutes from dilute aqueous samples by solid-phase extraction (SPE). Quantitative measurement of the extracted solutes was achieved by diffuse-reflectance spectroscopy (DRS) directly on the surface of the disk.     

Detection of propranolol in pharmaceutical formulations by diffuse reflectance spectroscopy

This paper describes an analytical reflectometric method that has an objective not only the industrial quality control but also to detect possible falsifications and/or adulterations of propranolol in pharmaceutical formulations. The method is based on the diffuse reflectance measurements of the colored product (III) of the spot test reaction between propranolol hydrochloride (I) and 2,6-dichloroquinone-4-chloroimide (II) using filter paper as solid support. Spot test conditions have been investigated using experimental design in order to identify and optimize the critical factors. The factors evaluated were DCQ concentration, propranolol solvent and DCQ solvent. The best reaction conditions were achieved with the addition of 30 microL of propranolol solution in ethanol 35% (v/v) and 30 microL of DCQ solution at 70 mg mL(-1) in acetone, in this order. All reflectance measurements were carried out at 500 nm and the linear range was from 8.45x10(-4) to 8.45x10(-2)mol L(-1) (r=0.998). The limit of detection was 1.01x10(-4)mol L(-1). No interference was observed from the assessed excipients and drugs. The method was applied to determine propranolol in commercial brands of pharmaceuticals. The results obtained by the proposed method were favorably compared with those given by the British Pharmacopoeia procedure.     

Comparison of calibration methods in quantitative diffuse reflectance infrared spectroscopy

Four different calibration methods were used for quantitative analysis of quartz and calcite in atmospheric aerosols by diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS): (A) conventional calibration with one measurement on each standard (single calibration); (B) calibration with an internal standard; (C) calibration with parallel (n = 4) measurements on each standard (multiple calibration); (D) multiple calibration followed by reference reflectance correction. The accuracy and the precision of the methods were compared and it was found that by using method D the reliability of the conventional pellet preparation transmission technique can be achieved.     

Determination of furosemide in pharmaceutical formulations by diffuse reflectance spectroscopy

In this report an analytical method to determine furosemide by using diffuse reflectance spectroscopy is presented. This study shows that this technique can give quantitative results using spot test analysis, particularly in the case of pharmaceuticals containing furosemide. The color spot test could be obtained by reaction between furosemide with p-dimethylaminocinnamaldehyde, in acid medium. This reaction produced a stable complex on filter paper after heating to 80 °C for 5 min. All reflectance measurements were carried out at 585 nm and the linear range was from 7.56×10⁻³ to 6.05×10⁻² mol l⁻¹, with a correlation coefficient of 0.999. The limit of detection was estimated to be 2.49×10⁻³ mol l⁻¹ (R.S.D.=1.7%) and the effect of common excipients on the reflectance measurements was evaluated. The method was applied to determine furosemide in commercial brands of pharmaceuticals. The results obtained by the proposed method were favorably compared with those of the official method, showing for the first time ever that quantitative spot test analysis by diffuse reflectance could be successfully used to determine furosemide in tablets.     

Element content in a number of plant leaves and accumulation of some elements in typical plant species: A case of Okayama Prefecture

Content of 40 elements in plant leaves, soils and rocks collected from several sampling sites in Okayama Prefecture was analyzed by instrumental neutron activation analysis using the Research Reactor of Kyoto University. Of these, 16 elements were taken up, and the relationship among their amounts in plants, rocks and soils was discussed, focusing mainly on the essential elements of higher plants and related elements in the periodic table. Although most element contents in plant were different among species, the mediam values of the most non-essential elements in the leaf were lower than the contents in the soil and rock samples. However, essential elements showed significantly higher median values than those in the soil and rock samples. Most element contents in leaves showed a logarithmic normal distribution. The normal distribution was observed in most essential elements except for zinc, while non-essential elements did not show a normal distribution. The results obtained by analysis of variance, showed that the plant species studied could be divided into four major groups, namelyDicotyledoneae, Monocotyledoneae, Gymnospermae andPteridophyta. In general,Dycotyledoneae andPteridophyta had higher element contents thanMonocotyledonaeae andGymnospermae.