Analytical performance of Raman spectroscopy in assaying biochemical components in human serum.

Chronic non-infectious diseases are important to research as they are the main causes of death in Brazil and worldwide. One very important chronic non-infectious disease is cardiovascular disease, whose risk factors (diabetes, dyslipidemia, and renal failure) can be detected through assessments of serum biochemical components. The objective of this study was to evaluate the analytical performance of Raman spectroscopy for analysis of lipid profile (total cholesterol, triglycerides, and HDL cholesterol), non-protein nitrogenous compounds (urea and creatinine), and glucose in 242 human serum samples. Models to discriminate and quantify the samples were developed using the predicted concentration by quantitative regression model based on partial least squares (PLS). The analytical error for the "leave-one-out" cross-validation based on the predicted PLS concentration was 10.5 mg/dL for total cholesterol, 21.4 mg/dL for triglyceride, 13.0 mg/dL for HDL cholesterol, 4.9 mg/dL for urea, 0.21 mg/dL for creatinine, and 15.4 mg/dL for glucose. The Kappa coefficient indicate very good agreement for cholesterol (0.83), good for triglyceride (0.77), urea (0.70) and creatinine (0.66), and fair for HDL cholesterol (0.38) and glucose (0.30). The results of the analytical performance demonstrated that Raman spectroscopy can be considered an important methodology to screen the population, especially for serum triglycerides and cholesterol.

Biochemical characterization of pathogenic bacterial species using Raman spectroscopy and discrimination model based on selected spectral features.

This study aimed to evaluate the differences in the Raman spectra of nine clinical species of bacteria isolated from infections (three Gram-positive and six Gram-negative species), correlating the spectra with the chemical composition of each species and to develop a classification model through discriminant analysis to categorize each bacterial strain using the peaks with the most significant differences. Bacteria were cultured in Mueller Hinton agar and a sample of biomass was harvested and placed in an aluminum sample holder. A total of 475 spectra from 115 different strains were obtained through a dispersive Raman spectrometer (830 nm) with exposure time of 50 s. The intensities of the peaks were evaluated by one-way analysis of variance (ANOVA) and the peaks with significant differences were related to the differences in the biochemical composition of the strains. Discriminant analysis based on quadratic distance applied to the peaks with the most significant differences and partial least squares applied to the whole spectrum showed 89.5% and 90.1% of global accuracy, respectively, for classification of the spectra in all the groups. Raman spectroscopy could be a promising technique to identify spectral differences related to the biochemical content of pathogenic microorganisms and to provide a faster diagnosis of infectious diseases.

Quantifying glucose and lipid components in human serum by Raman spectroscopy and multivariate statistics.

Raman spectroscopy has been employed in the quantitative analysis of biochemical components in human serum. This study aimed to develop a spectral model to estimate the concentration of glucose and lipid fractions in human serum, thus evaluating the feasibility of Raman spectroscopy technique for diagnostic purposes. A total of 44 samples of blood serum were collected from volunteers submitted to routine blood biochemical assay analysis. The biochemical concentrations of glucose, triglycerides, cholesterol, and high-density and low-density lipoproteins (HDL and LDL) were obtained by colorimetric method. Serum samples (200 µL) were submitted to Raman spectroscopy (830 nm, 250 mW, 50-s accumulation). The spectra of sera present peaks related to the main constituents, particularly proteins and lipids. A quantitative model based on partial least squares (PLS) regression has been developed to estimate the concentration of these compounds, taking the biochemical concentrations assayed by the colorimetric method as sample's actual concentrations. The PLS model based on leave-one-out cross-validation approach estimated the concentration of triglycerides and cholesterol with r = 0.98 and 0.96, and root mean square error of 35.4 and 15.9 mg/dL, respectively. For the other biochemicals, the r was ranging from 0.75 to 0.86. These results evidenced the possibility of performing biochemical assay in blood serum samples by Raman spectroscopy and PLS regression and may be employed as a means of diagnosis in routine clinical analysis.

Discrimination of selected species of pathogenic bacteria using near-infrared Raman spectroscopy and principal components analysis.

ABSTRACT. A method, based on Raman spectroscopy, for identification of different microorganisms involved in bacterial urinary tract infections has been proposed. Spectra were collected from different bacterial colonies (Gram-negative: Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa and Enterobacter cloacae, and Gram-positive: Staphylococcus aureus and Enterococcus spp.), grown on culture medium (agar), using a Raman spectrometer with a fiber Raman probe (830 nm). Colonies were scraped from the agar surface and placed on an aluminum foil for Raman measurements. After preprocessing, spectra were submitted to a principal component analysis and Mahalanobis distance (PCA/MD) discrimination algorithm. We found that the mean Raman spectra of different bacterial species show similar bands, and S. aureus was well characterized by strong bands related to carotenoids. PCA/MD could discriminate Gram-positive bacteria with sensitivity and specificity of 100% and Gram-negative bacteria with sensitivity ranging from 58 to 88% and specificity ranging from 87% to 99%.

Detecting alterations of glucose and lipid components in human serum by near-infrared Raman spectroscopy

Introduction Raman spectroscopy may become a tool for the analysis of glucose and triglycerides in human serum in real time. This study aimed to detect spectral differences in lipid and glucose components of human serum, thus evaluating the feasibility of Raman spectroscopy for diagnostic purposes. Methods A total of 44 samples of blood serum were collected from volunteers and submitted for clinical blood biochemical analysis. The concentrations of glucose, cholesterol, triglycerides, and low-density and high-density lipoproteins (LDL and HDL) were obtained using standard biochemical assays. Serum samples were placed in Eppendorf tubes (200 µL), kept cooled (5 °C) and analyzed with near-infrared Raman spectroscopy (830 nm, 250 mW, 50 s accumulation). The mean spectra of serum with normal or altered concentrations of each parameter were compared to determine which Raman bands were related to the differences between these two groups. Results Differences in peak intensities of altered sera compared to normal ones depended on the parameter under analysis: for glucose, peaks were related to glucose; for lipid compounds the main changes occurred in the peaks related to cholesterol, lipids (mainly triolein) and proteins. Principal Components Analysis discriminated altered glucose, cholesterol and triglycerides from the normal serum based on the differences in the concentration of these compounds. Conclusion Differences in the peak intensities of selected Raman bands could be seen in normal and altered blood serum samples, and may be employed as a means of diagnosis in clinical analysis.