ABSTRACT
The baseline drifts of Raman spectra occur in many types of instrumental measurements. It is an important part and a routine step to correct the baseline drift for the data preprocessing. In the present work, the limitations of the baseline correction method based on polynomial fitting were highlighted and a modified polynomial fitting method, i. e. piecewise linear fitting method, was proposed. Combined with the computer, this method could eliminate the baseline automatically. A series of Raman spectra of single polystyrene bead, red blood cell or yeast cell acquired by laser tweezers Raman spectroscopy were preprocessed by this method and its efficiency was verified. The results demonstrated that piecewise linear fitting can correct the baseline shifts effectively and provides more accurate information for further data analysis. It is a feasible method for correction of Raman spectrum.
Subject(s)
Algorithms , Artifacts , Models, Statistical , Spectrum Analysis, Raman/methods , Regression AnalysisABSTRACT
The pigments from Rhodotorula glutinis were separated by using thin layer chromatography, and the result showed that Rhodotorula glutinis cells could synthesize at least three kinds of pigments, which were beta-carotene, torulene, and torularhodin. The Raman spectra based on the three pigments were acquired, and original spectra were preprocessed by background elimination, baseline correction, and three-point-smoothing, then the averaged spectra from different pigments were investigated, and the result indicated that Raman shift which represents C-C bond was different, and the wave number of beta-carotene demonstrated the largest deviation, finally torulene and torularhodin in Rhodotorula glutinis had more content than beta-carotene. Quantitative analysis of Raman peak height ratio revealed that peak height ratio of pigments showed little difference, which could be used as parameters for further research on living cells, providing reference content of pigments. The above results suggest that Raman spectroscopy combined with thin layer chromatography can be applied to analyze pigments from Rhodotorula glutinis, provides abundant information about pigments, and serves as an effective method to study pigments.
Subject(s)
Chromatography, Thin Layer , Rhodotorula/chemistry , Spectrum Analysis, Raman , beta Carotene/analysis , Carotenoids/analysisABSTRACT
In the present paper, a method was established based on laser tweezer Raman spectroscopy for rapid quantification of astaxanthin in Phaffia rhodozyma cells. First, the Raman spectra of astaxanthin standard solution with different concentrations were determined and the standard curve for astaxanthin with the peak intensity at 1 520 cm- was plotted; And then the Phaffia yeast cells cultivated in different nitrogen source and carbon source medium were divided into two parts, one for the detection of Raman spectra, and the other for the determination of ultraviolet visible spectrophotometry; Finally the relationship between the two methods was analyzed. The correlation coefficient of standard curve for astaxanthin is 0.998 3. Comparing laser tweezers Raman spectroscopy method with traditional ultraviolet visible spectrophotometry in analyzing the content of astaxanthin in unit mass Phaffia rhodozyma and the yield of astaxanthin in unit volume fermentation broth of Phaffia rhodozyma, the authors found that the data obtained have good linear relationship. And the correlation coefficients are 0.917 7 and 0.905 4, respectively. Therefore, both methods have almost the same effect of measuement. But laser tweezers Raman spectroscopy method is more efficient in the quantitative analysis of astaxanthin in Phaffia rhodozyma cells.
Subject(s)
Basidiomycota/chemistry , Optical Tweezers , Spectrum Analysis, Raman , Carbon , Fermentation , Nitrogen , Xanthophylls/analysisABSTRACT
Carotenoid synthesis in Rhodotorula glutinis was investigated with Raman tweezers in order to find the effect of nitrogen and carbon resource on carotenoid yield. The cells in fermentation terminus were harvested, and then divided into two parts, one for UV analysis, the other for Raman tweezers detection. Original spectra were preprocessed by carrying out background elimination and baseline correction, and the averaged spectra of cells cultivated in different fermentation medium were analyzed qualitatively. The results showed that the Raman intensity of carotenoid were obviously different. There was a high correlation between UV results and Raman peak height data, the correlation coefficients of fitted parameters were 0.907 8 and 0.912 1, respectively. Quantitative analysis of 1 508 cm(-1) peak height indicated that the appropriate nitrogen and carbon resources for the growth of Rhodotorula glutinis cells and synthesis of carotenoid were yeast extract + tryptone, and glucose, respectively. The above results suggest that Raman tweezers can provide information about carotenoids in Rhodotorula glutinis cells and serve as an effective tool for real time measurement of carotenoid synthesis and optimization of fermentation medium.
Subject(s)
Carotenoids/biosynthesis , Rhodotorula/chemistry , Carbon , Fermentation , Glucose , Nitrogen , Optical Tweezers , Spectrum Analysis, RamanABSTRACT
Fourier transform infrared spectrometer was used to collect infrared spectra of Cortex Phellodendri from six different regions. Original spectra were preprocessed by carrying out appropriate baseline correction and five-points smoothing, and the averaged spectra of Cortex Phellodendri from the six origins were analyzed. As a result, the averaged spectra looked quite similar. The normalized spectra were selected to construct principal component analysis model in the range of fingerprint region 1800 - 500 cm(-1), and according to the model, the first three principal components accounted for 98% of the variance information in the fingerprint region, and each sample was able to form distinct cluster in the principal component space, then the identification of Cortex Phellodendri from the six regions was basically achieved; besides, to some extent, the sparse density of the samples distribution reflected the genetic relationship. The loading factors of the model were analyzed, and the results indicated that the differences between Cortex Phellodendri samples mostly depended on the contents of protein, carbohydrates, lipids, alkaloids, sterols, obaculactone, oba-cunone, and obacunonlc acid. On the whole, combined with principal component analysis, FTIR provides an effective way to evaluate the herbal Cortex Phellodendri rapidly and nondestructively, which also reflects the content difference of material composition.
Subject(s)
Carbohydrates/analysis , Phellodendron/chemistry , Plant Proteins/analysis , Spectroscopy, Fourier Transform Infrared , Principal Component AnalysisABSTRACT
Apoptosis of gastric cancer cells induced by cisplatin was investigated using laser Raman spectroscopy. Gastric cancer cells (SGC7901) were treated with 10 microg x mL(-1) cisplatin for 24, 48 and 72 hours, then were divided into two parts, one for fluorescence staining, the other for collection of Raman spectra by means of scanning. The acquired spectra were then preprocessed by background elimination, smoothing, normalization, baseline correction, and peak fitting. Fluorescence staining result showed that the nucleuses from untreated group were uniformly stained, while those from the group treated for 72 hours were densely stained and broken. The spectra results revealed that the intensity of peaks associated with nucleic acid and protein decreased after the cells were incubated with cisplatin for 24, 48 and 72 hours. The intensity of peaks at 783, 1002 and 1343 cm(-1) respectively fell to 52, 64 and 76 percent of the original value after 72 h of treatment, which indicated that cisplatin could induce apoptosis of gastric cancer cells and reduce the amount of nucleic acid and protein in the cells. The above results suggest that Raman spectra can provide abundant information about the changes in materials in cells and serve as an effective method for real time measurement of apoptosis.