Application of Legendre Polynomials Correction Before Component Quantification of Biological Mid-infrared Spectra

Abstract

Quantitative determination from infrared spectra still faces problems that are not completely resolved to date. One of these problems is the deformation of baselines.

We have tested the Legendre baseline correction method by subtracting from a spectrum the low-degreed terms from a Legendre polynomial function which is obtained by the decomposition of the spectrum.

As opposed to the filtered spectra with n=0 which remained very similar to the original spectra, the filtered spectra, as from n=2, exhibit a great amount of distorsion. This result suggests that it is as from n=2, that the terms that are meaningful to the signal (not just to baseline), are to be substracted.

Mathematical methods such as Principal Component Analysis and Principal Component Regression are used to analyse quantitatively components from biological samples. The thus predicted sucrose content values for values of n that range between 0 and 8 (n={0, 2, 4, 6, 8}) show that the best results are obtained for n=4. The mean and standard deviation values of the difference between reference and predicted values varies between 8.5 × 10^?2 and 6.6 × 10^?2 and between 0. 224 and 0. 195 respectively; these values are minimum when the n=4 term is subtracted from the spectra.

This processing hence, sensibly improves the quantification of components from Mid-FTIR spectra of biological solutions. This procedure would be interesting in cases where the precision of quantitative measures is essential.     

傅里叶变换红外光谱结合化学计量学鉴别预包装纯菠萝汁中外源糖

为了快速、准确鉴别预包装纯菠萝汁是否掺有外源性糖,采用傅里叶变换衰减全反射中红外光谱技术采集900～1 500 cm-1范围内不同批次的预包装纯菠萝汁样品和掺入甜菜糖浆、大米糖浆、木薯糖浆的菠萝汁掺假样品的中红外光谱共计234例,以线性判别分析和支持向量机分析为掺假鉴别模型的建模方法,比较了全波段谱图与通过主成分载荷系数分析选取特征波长图谱的两种掺假鉴别模型。研究表明,全波段图谱的线性判别分析和支持向量机分类模型对验证集的判断正确率均高于88%;选取8个特征波长之后,线性判别分析模型验证集判断正确率提高至96.15%,支持向量机模型验证集判断正确率提高至94.87%,且模型输入变量由312个减少到8个。利用傅里叶变换衰减全反射中红外光谱技术结合化学计量学方法选取特征波长后建立的模型可以较好的应用于预包装纯菠萝汁外源糖的鉴别。     

α-NH2 Determination from Biological Infrared Spectra

The use of Mid-infrared spectroscopy for the analysis of food products has only recently developped with the advent of Fourier Transformed spectroscopy and other techniques like Attenuated Total Reflectance, diffused reflection combined to the use of powerful micro-computers. We have recently reported to use a combination of multidimensional statistical analysis and Mid-FTIR spectroscopy for the quantitative determination of sugars in a biological sample. In this paper we have evaluated the use of such a method for the quantitative measurement of α-amino groups from ammo-acids, peptides and proteins. The spectral region where the characteristic absorption bands of such groups are located, ranges between 1200 cm-1 and 1900 cm-1. Water features a major absorption band in this region (1500–1700 cm-1). This superimpose with amide I and II bands.

The standard deviation for each and every wavelength, calculated for all the spectra of the calibration set, show the existence of two absorption bands in the 1500–1900 cm-1 region which means that the observed variations in this zone are not only due to water but are also due to two peaks centered at 1650 cm-1 and 1540 cm-1 (with a hollow at 1600 cm-1) that are characteristic of protids.

The contribution of the first four axes of the PCA, axes 5, 1, 4 and 2, to the total inertia percentage are 2. 37%, 53. 36%, 3. 92% and 28. 82% respectively. The correlation coefficient between the major axis, axis 5, and the chemical values of α-NH₂ is 0. 311 and the second axis, axis 1, increases this value to 0. 541. The first 10 axes were used to establish the prediction equation; the correlation coefficient value is very high: 0. 978.

Good predictions were obtained; mean and standard deviation associated to the predicted concentrations of α-NH₂content, valued 0 g/ml and 0. 12 g/ml respectively. Hence, we have established the possibility of determining, from a MIR spectra, the α-NH₂ content.     

Gram-Schmidt Orthogonalization and Elimination of the Effect of Unwanted Component Spectra Applied to a Biological Mid-infrared Spectra Collection

Abstract

Near Infrared spectroscopy (NIR) is the most widely used technique for the analysis of major biochemical constituents in food products. The Mid-infrared range spectroscopy is also being more and more studied in the field of food analysis. This range was primarily applied to qualitative analysis of food components, but with the advent of new techniques such as Attenuated Total Reflectance (ATR) together with the possibility of combination with powerful micro-computers, MIR is now more and more used for quantitative analysis.

In addition to baseline deformations, interference by unwanted compounds are major sources of problems that are encountered in analyses.

We have previously proposed (Cadet et al., 1991) the use of multidimensional statistical analysis combined with Mid-FTIR spectroscopy for the prediction of sucrose concentrations in biological solutions containing three sugars: sucrose, fructose and glucose.

In this paper, a least-squares method has been used to assess the elimination of the component spectra associated to the fructose (the unwanted components were first orthogonal zed and normalized by the Gram-Schmidt orthogonalization method). This procedure permitted the automatic subtraction of discriminant spectral patterns representative of fructose concentration before application of Principal Component Analysis (PCA) and Principal Component Regression (PCR). PCA was applied independently before and after spectral correction of the collections. It is found that when the factorial maps obtained before and after correction are compared, the elimination procedure improves significantly the classification of solutions according to their sucrose content. However the bias and standard deviation (SD) values that are associated with the sucrose content predicted values are not influenced by the correction method used: bias and SD are 3.62×10^?2 and 3.097×10^?1 before correction and after correction they were respectively 3.60×10^?2 and 3.104×10^?1. This could be explained by the fact that the presence of fructose in solution does not interfere with caracteristic absorption bands of sucrose and that sucrose and fructose concentrations are strongly correlated. The absorbtions bands of the spectral representation of the principal component, which is identified to be that associated with sucrose, are identical before and after correction.     

INDIRECT METAL ION (K+, NA+, MG2+, AND CA2+) QUANTIFICATION FROM INFRARED SPECTROSCOPY

ABSTRACT

In this paper, we report the use of Mid-FTIR spectroscopy coupled with Partial Least Squares method for the quantitative determination of various alkaline and alkaline earth metals in aqueous solution owing to their interactions with sucrose. First of all, prediction equations that linked cation concentration to the spectral data were established independently for each ion (K⁺, Na⁺, Mg²⁺ or Ca²⁺): very high correlation coefficient values between the two first axes and the chemical values were obtained. Moreover, a good prediction could be made whatever the nature of ion involved in interaction with sucrose. Then, all spectral data have been gathered for generating a common prediction equation. In this case, the predictions of metal ion concentration are almost as much accurate. For both regression models, Mg²⁺ appears to provide the best precision in quantification. Nevertheless, the different types of aqueous solution, regarding to ions, can be discriminated on the basis of their spectral data set up with the three mostly correlated axes of the PCA.     

Evidence for Potassium-Sucrose Interaction in Biological Mid-Infrared Spectra by Multidimensional Analysis

Complex-formation between carbohydrates and cations could have important biological implications.

In this work, Mid-Infrared spectra of pure sucrose solutions and of biological solutions containing sucrose and potassium ions (K+) were investigated by Principal Component Analysis (PGA).

By direct examination of the Mid-Infrared spectra of the biological solutions containing K⁺ ions, no interactions between the cations and sucrose molecules could be observed. However, when the spectral pattern obtained by PCA and which is associated with sucrose, was examined, splitting and shifts in the characteristic absorption bands were observed owing to interactions between sucrose molecules and K⁺ ions. The 997 cm-1 peak which had a visible shoulder at 991 cm-1 and that is observed in pure solutions, was decomposed in the biological solutions into 3 distinct peaks at 1004, 996 and 990 cm-1. The two peaks centered at 1053 cm-1 were split into 3 peaks: 1060, 1051, 1045 cm-1. Hence by PCA, shoulders were characterized in biological solutions and more distinct peaks could be observed. These split and shift phenomena are similar to those obtained when crystalline sugar salts were investigated. This type of interaction, involving potassium ions and sucrose molecules, would be responsible for the storage of this cation which role is essential in plant metabolism.     

Monitoring radiation induced alterations in biological systems,from molecules to tissues,through infrared spectroscopy

Humans can be exposed to non-ionizing and ionizing radiation for diagnostic, therapeutic, accidental, and occupational reasons. Consequently, the effect of radiation on biological systems has attracted the attention of researchers for a rather long time. This review is about the mid-infrared Fourier Transform Infrared (FTIR) spectroscopic characterization of non-ionizing and ionizing radiation-induced changes in DNA, lipids, and proteins, as isolated or synthetic macromolecules, and in biological membranes, cells, and tissues. Here, the context of radiation was limited with electromagnetic radiation including gamma rays. The review first outlines introductory information about non-ionizing and ionizing radiation and their interaction with biological systems. Afterwards, FTIR spectroscopy and spectroscopic analysis are briefly discussed. Finally, FTIR spectroscopic analysis of DNA, lipids, proteins, membranes, cells, and tissues that were exposed to radiation are presented. The findings show that FTIR spectroscopy can be successfully used as a novel method to monitor radiation-induced alteratios in biological systems.     

Non-invasive identification of surface materials on marble artifacts with fiber optic mid-FTIR reflectance spectroscopy

Research work using a compact and portable fiber optic mid-infrared reflectance spectrometer has been carried out to study surface materials on marble, first on laboratory reference models and then on historical objects. The laboratory research has shown that the complex optical reflectance phenomena of layered samples can be interpreted in terms of the nature of the compounds present (sulfates, oxalates, phosphates, resins, waxes and proteins were studied) even with a very low signal overlapping with the strong carbonate reflectance. The portable instrument was used for in situ examination of the surface condition of two Italian marble works of art: the Deposizione dalla Croce by Benedetto Antelami in the Parma cathedral and the David by Michelangelo in the Galleria dell’Accademia in Firenze.