Identification and determination of different processed products and their extracts of Crataegi Fructus by infrared spectroscopy combined with two-dimensional correlation analysis.

The fruit of Crataegus sp. is known as "Shanzha (SZ)" in China and is widely used in the food, beverage, and traditional Chinese medicine (TCM) industries. SZ usually requires thermal processing to reduce the irritation of its acidity to the gastric mucosa. Different processed products of SZ resulting from thermal processing have different or even opposite functions in clinical applications. In addition, 5-hydroxymethylfurfural (5-HMF) intermediates produced during thermal processing are carcinogenic to humans. Therefore, the aim of this study was to explore a rapid and accurate method by Fourier transform infrared spectroscopy (FT-IR) for the identification of different processed products and the determination of 5-HMF in extracts. In qualitative identification, a three-stage infrared spectroscopy identification method (raw spectra, the second derivative spectra, and two-dimensional correlation (2DCOS) spectra) was developed to distinguish different processed products of SZ step by step. In quantitative determination, partial least squares regression combined with different variable selection methods, especially the 2DCOS method, was applied to determine the 5-HMF content. The results show that temperature-induced 2DCOS synchronous spectra can effectively identify different processed products of SZ by shape, intensity, and position of auto-peaks or cross-peaks, and the variables selected by power spectra from concentration-induced 2DCOS synchronous spectra have better prediction ability for 5-HMF compared to full variables. The above results demonstrate that 2D-COS analysis is a potential tool in qualitative and quantitative analysis, which can improve sample identification accuracy and determination capabilities. This study not only establishes a rapid and accurate method for the identification of different processed products but also provides a practical reference for food safety and the efficient use of TCM.

Usefulness of mid-infrared spectroscopy as a tool to estimate body condition score change from milk samples in intensively fed dairy cows.

Directly measuring individual cow energy balance is not trivial. Other traits such as body condition score (BCS) and BCS change (ΔBCS) can, however, be used as an indicator of cow energy status. Body condition score is a metric used worldwide to estimate cow body reserves, but the estimation of ΔBCS was, until now, conditional on the availability of multiple BCS assessments. The aim of the present study was to estimate ΔBCS from milk mid-infrared (MIR) spectra and days in milk (DIM) in intensively fed dairy cows using statistical prediction methods. Daily BCS was interpolated from cubic splines fitted through the BCS records and daily ΔBCS was calculated from these splines. The ΔBCS records were merged with milk MIR spectra recorded on the same week. The dataset comprised 37,077 ΔBCS phenotypes across 9,403 lactations from 6,988 cows in 151 herds based in Quebec, Canada. Partial least squares regression (PLSR) and a neural network (NN) were then used to estimate ΔBCS from (1) MIR spectra only, (2) DIM only, or (3) MIR spectra and DIM together. The ΔBCS data in both the first 120 and 305 DIM of lactation were used to develop the estimates. Daily ΔBCS had a standard deviation of 4.40 × 10-3 BCS units in the 120-d dataset and of 3.63 × 10-3 BCS units in the 305-d dataset. A 4-fold cross-validation was used to calibrate and test the prediction equations. External validation was also conducted using more recent years of data. Irrespective of whether based on the first 120 or 305 DIM, or when MIR spectra only, DIM only or MIR spectra and DIM were jointly used as prediction variables, NN produced the lowest root mean square error (RMSE) of cross-validation (1.81 × 10-3 BCS units and 1.51 × 10-3 BCS units, respectively, using the 120-d and 305-d dataset). Relative to predictions for the entire 305 DIM, the RMSE of cross-validation was 15.4% and 1.5% lower in the first 120 DIM when using PLSR and NN, respectively. Predictions from DIM only were more accurate than those using just MIR spectra data but, irrespective of the dataset and of the prediction model used, combining DIM information with MIR spectral data as prediction variables reduced the RMSE compared with the inclusion of DIM alone, albeit the benefit was small (the RMSE from cross-validation reduced by up to 5.5% when DIM and spectral data were jointly used as model features instead of DIM only). However, when predicting extreme ΔBCS records, the MIR spectral data were more informative than DIM. Model performance when predicting ΔBCS records in future years was similar to that from cross-validation demonstrating the ability of MIR spectra of milk and DIM combined to estimate ΔBCS, particularly in early lactation. This can be used to routinely generate estimates of ΔBCS to aid in day-to-day individual cow management.

Neuroprotective constituents of Actaea acuminata (Wall. ex Royle) H. Hara roots.

The methanol extract and its ethyl acetate fraction (EAF) of Actaea acuminata (Wall. ex. Royle) H. Hara roots were reported to exhibit significant antianxiety, anticonvulsant and antidepressant activities, and mild sedative activity. But the constituents responsible for these activities have not been isolated. The present study was undertaken to isolate neuroprotective compounds of A. acuminata following bioactivity-guided-fractionation. The column chromatography of EAF and its sub-fractions led to the isolation of four phenolic compounds (bergenin, gallic acid, acetyl bergenin and racemic mixture of diacetyl bergenin), which were characterized by IR and NMR spectral analysis. All the compounds exhibited significant antianxiety and antidepressant activities with respect to control. The gallic acid and bergenin did not show anticonvulsant activity, whereas acetyl bergenin and racemic mixture of diacetyl bergenin exhibited significant anticonvulsant activity. Neuropharmacological activities of A. acuminata are attributed due to polyphenolic compounds. Scientific validation of traditional claims of A. acuminata has opened up roadmap of research for the development of CNS affecting lead molecules.

Difference in the metabolic characteristics of chronic obstructive pulmonary disease patients and healthy adults.

INTRODUCTION: By detecting the metabolic difference of the Heart and Lung meridians, the present study aims to investigate the specificity of different meridians and verify whether functional near infrared spectroscopy is validated as an add-on technique to assist diagnosis of chronic obstructive pulmonary disease (COPD). METHODS AND ANALYSIS: The Lung and Heart meridians are chosen as the target for comparison; accordingly, 120 eligible participants will be included and divided into the COPD group, healthy control group, and healthy intervention group. Functional near infrared spectroscopy will be adopted to measure the metabolic characteristics of the Heart and Lung meridians. On one hand, the specificity of the meridian-visceral association will be investigated by comparing the metabolic difference in the Heart and Lung meridians between the healthy control group and COPD group. On the other hand, the specificity of site-to-site association will be determined by comparing the metabolic change between the 2 meridians that induced by moxibustion in the Heart meridian and Lung meridian, respectively, in the healthy control group. The primary outcome will be regional oxygen saturation of corresponding regions along the Heart and Lung meridians. TRIAL REGISTRATION: ClinicalTrials.gov NCT04046666.

Vibrational Spectroscopic Based Approach for Diagnosing Babesia bovis Infection.

Babesia bovis parasites present a serious and significant health concern for the beef and dairy industries in many parts of the world. Difficulties associated with the current diagnostic techniques include the following: they are prone to human error (microscopy) or expensive and time-consuming (polymerase chain reaction) to perform. Little is known about the biochemical changes in blood that are associated with Babesia infections. The discovery of new biomarkers will lead to improved diagnostic outcomes for the cattle industry. Vibrational spectroscopic technologies can record a chemical snapshot of the entire organism and the surrounding cell thereby providing a phenotype of the organism and the host infected cell. Here, we demonstrate the applicability of vibrational spectroscopic imaging techniques including Atomic Force Microscopy Infrared (AFM-IR) and confocal Raman microscopy to discover new biomarkers for B. bovis infections. Furthermore, we applied Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR) to detect B. bovis in red blood cells (RBCs). Based on changes in the IR spectral bands, with ATR-FTIR in combination with Partial Least Squares-Discriminant Analysis we were able to discriminate infected samples from controls with a sensitivity and specificity of 92.0% and 91.7%, respectively, in less than 2 min, excluding sample extraction and preparation. The proposed method utilized a lysis approach to remove hemoglobin from the suspension of infected and uninfected cells, which significantly increased the sensitivity and specificity compared to measurements performed on intact infected red blood cells (intact infected RBC, 77.3% and 79.2%). This work represents a holistic spectroscopic study from the level of the single infected RBC using AFM-IR and confocal Raman to the detection of the parasite in a cell population using ATR-FTIR for a babesiosis diagnostic.

Improved infrared spectroscopic discrimination between gall bladder (GB) polyps and GB cancer using component-descriptive spectral features of separated phases from bile.

This study demonstrates a unique strategy for enhancing infrared (IR) spectroscopic discrimination between gall bladder (GB) polyps and cancer. This strategy includes the separation of raw bile juice into three sections of organic, aqueous, and amphiphilic phases and a cooperative combination of all IR spectral features of each separated phase for the discrimination. Raw bile juice is viscous and complex in composition because it contains fatty acids, cholesterol, proteins, phospholipids, bilirubin, and other components; therefore, the acquisition of IR spectra providing more component-discernible information is fundamental for improving discrimination. For this purpose, raw bile juice was separated into an aqueous phase, mostly containing bile salts, an organic phase with isolated lipids, and an amphiphilic phase, mainly containing proteins. The subsequent IR spectra of each separated phase were mutually characteristic and complementary to each other. When all the IR spectral features were combined, the discrimination was improved compared to that using the spectra of raw bile juice with no separation. The cooperative integration of more component-specific spectra obtained from each separated phase enhanced the discrimination. In addition, the IR spectra of the major constituents in bile juice, such as bile acids, conjugated bile salts, lecithin, and cholesterol, were recorded to explain the IR features of each separated phase.

Rapid prediction of yellow tea free amino acids with hyperspectral images.

Free amino acids are an important indicator of the freshness of yellow tea. This study investigated a novel procedure for predicting the free amino acid (FAA) concentration of yellow tea. It was developed based on the combined spectral and textural features from hyperspectral images. For the purposes of exploration and comparison, hyperspectral images of yellow tea (150 samples) were captured and analyzed. The raw spectra were preprocessed with Savitzky-Golay (SG) smoothing. To reduce the dimension of spectral data, five feature wavelengths were extracted using the successive projections algorithm (SPA). Five textural features (angular second moment, entropy, contrast, correlation, and homogeneity) were extracted as textural variables from the characteristic grayscale images of the five characteristic wavelengths using the gray-level co-occurrence matrix (GLCM). The FAA content prediction model with different variables was established by a genetic algorithm-support vector regression (GA-SVR) algorithm. The results showed that better prediction results were obtained by combining the feature wavelengths and textural variables. Compared with other data, this prediction result was still very satisfactory in the GA-SVR model, indicating that data fusion was an effective way to enhance hyperspectral imaging ability for the determination of free amino acid values in yellow tea.

Potentiality of front-face fluorescence and mid-infrared spectroscopies coupled with partial least square regression to predict lipid oxidation in pound cakes during storage.

The potentialities of front-face fluorescence (FFF) and mid-infrared (MIR) spectroscopies coupled with partial least square regression (PLSR) were compared to predict the lipid oxidation of pound cakes. The level of lipid oxidation in pound cakes determined using classical methods showed some changes. Similarly, the fluorescence emission (305-490 nm) and excitation (252-390 nm) spectra and MIR spectra scanned in the 4000-700 cm-1 region showed some changes in pound cakes as a function of both storage time and the type of oil used in the formulation. The application of PLSR to the MIR spectra, provided excellent predictive results for free fatty acid (R2 = 0.97) and peroxide values (R2 = 0.87). Similar results were obtained from both tryptophan and MIR spectra for the prediction of TOTOX (R2 > 0.86) demonstrating the efficiency of the MIR and FFF spectroscopies to qualify and quantify the level of lipid oxidation in pound cakes.

Understanding the infrared and Raman spectra of ganoderic acid A: An experimental and DFT study.

Ganoderic Acids (GAs) are the major medicinal compounds in Ganoderma lucidum used as traditional Chinese medicine since ancient times. Ganoderic acid A (GAA) is the first discovered ganoderic acids reported in the literature, which is also one of most abundant triterpenoids of Ganoderma lucidum. Especially, GAA has been extensively investigated in recent decades for its positive medicinal activities. However, the vibrational properties of GAs have rarely been studied or reported. In this work, we focused on the typical GAA and studied the infrared (IR) and Raman spectra based on both experiments and DFT calculations. As such, we could not only achieve the assignments of the vibrational modes, but also from the IR and Raman spectra, we found that the spectral region from 1500 cm-1 to 1800 cm-1 is particularly useful for distinguishing different types of GAs. In addition, its dehydrogenated derivative ganoderenic acid A (GOA) was also studied, which could be identified due to its spectral feature of strong IR and Raman bands around 1620 cm-1. This work therefore may facilitate the application of IR and Raman spectroscopies in the inspection and quality control of Ganoderma lucidum.

Commonalities and characteristics of aqueous extracts from three Uighur medicines were analyzed by using three-stage infrared spectroscopy combined with ultra-performance liquid chromatography-time of flight-mass spectra.

OBJECTIVE: To establish a method to analyze the commonalities and characteristics of the aqueous extracts from three Uighur medicines. METHODS: In this study, a combination method of three-stage infrared spectroscopy and ultra-performance liquid chromatography-time of flight-mass spectra (UPLC-TOF-MS) method was used to analyze the commonalities and characteristics of the aqueous extracts from Hezi (Fructus Chebulae), Maohezi (Terminalia Belliricae Fructus) and Xiqingguo (Chebulae Fructus Immaturus). RESULTS: In Fourier transform-infrared spectroscopy spectra, all three samples showed the characteristic absorption peaks of tannins similarly. According to UPLC-TOF-MS data analysis, the difference of the position and intensity of the peaks at 1713- 1707 cm－1 in the three samples were related to the difference in the relative content of tannin and pentacyclic triterpenoids. In second derivative infrared spectroscopy spectra, further analysis of the differences in the infrared spectra of the three samples was performed by increasing the apparent resolution. In combination with UPLC-TOF-MS data, some features infrared absorption peaks were assigned. The absorption peaks at 1032-1030 cm－1 assigned to the stretching vibration of C-O-C (ether bond) groups of tannins; 1164-1163, 1063-1062, 1009- 1005, 904 cm－1 attributed to the stretching vibration of C-O (ether bond) and C-O-C groups of glycosidic compounds. 1385-1383 cm－1 attributed to the bending vibration of C-H (alkyl hydrogen) of methoxyl group of pentacyclic triterpenoids. Peak 835 cm－1 was assigned to the characteristic absorption peak of bending vibration of = C-H groups of pentacyclic triterpenoids. The two dimensional correlation infrared spectroscopy could further quickly distinguish three samples through the dynamic structural information of their chemical components and discrepancy of auto-peaks and cross- peaks intuitively in the range of 1720-419 cm－1. CONCLUSION: By comparing the intensity of the infrared characteristic absorption peaks, the main chemical components were assigned, which significantly enriched and perfected the data analysis of the infrared spectra of three Uighur Medicines. It provided a rationale for the identification of medicinal materials with complex and similar chemical components using the three-stage infrared spectroscopy and UPLC-TOF-MS.

Near-field infrared nanospectroscopy and super-resolution fluorescence microscopy enable complementary nanoscale analyses of lymphocyte nuclei.

Recent super-resolution fluorescence microscopy (3D-Structured Illumination Microscopy, 3D-SIM) studies have revealed significantly altered nuclear organization between normal lymphocyte nuclei and those of classical Hodgkin's Lymphoma. Similar changes have been found in Multiple Myeloma (MM) nuclei, as well as in a premalignant condition, Monoclonal Gammopathy of Unknown Significance (MGUS). Using 3D-SIM, an increase in DNA-poor and DNA-free voids was evident in reconstructed 3D-SIM images of diseased nuclei at 40 nm pixel resolution (x,y: 40 nm, z: 80 nm). At best, far-field FTIR imaging yields spatially resolved images at ∼500 nm spatial resolution; however, near-field infrared imaging breaks the diffraction limit at a scale comparable to that of 3D-SIM, providing details on the order of 30 nm spatial resolution. We report here the first near-field IR imaging of lymphocyte nuclei, and far-field IR imaging results of whole lymphocytes and nuclei from normal human blood. Cells and nuclei were mounted on infrared-compatible substrates, including CaF2, undoped silicon wafers, and gold-coated silicon wafers. Thermal source far-field FTIR images were obtained with an Agilent-Cary 620 microscope, 15× objective, 0.62 NA and 64 × 64 array Focal Plane Array detector (University of Manitoba), or with a similar microscope equipped with both 15× and 25× (0.81 NA) objectives, 128 × 128 FPA and either thermal source or synchrotron source (single beam) infrared light at the Advanced Light Source (ALS), LBNL, Berkeley CA. Near-field IR spectra were acquired at the ALS, on the in-house SINS equipment, as well as with a Neaspec system, both illuminated with synchrotron light. Finally, some near-field IR spectra and images were acquired at Neaspec GmbH, Germany. Far-field IR spectra of normal cells and nuclei showed the characteristic bands of DNA and proteins. Near-field IR spectra of nuclei showed variations in bands assigned to protein and nucleic acids including single and double-stranded DNA. Near-field IR images of nuclei enabled visualization of protein and DNA distribution in spatially-resolved chromosome territories and nuclear pores.

w-Type ions formed by electron transfer dissociation of Cys-containing peptides investigated by infrared ion spectroscopy.

In mass spectrometry-based peptide sequencing, electron transfer dissociation (ETD) and electron capture dissociation (ECD) have become well-established fragmentation methods complementary to collision-induced dissociation. The dominant fragmentation pathways during ETD and ECD primarily involve the formation of c- and z• -type ions by cleavage of the peptide backbone at the N─Cα bond, although neutral losses from amino acid side chains have also been observed. Residue-specific neutral side chain losses provide useful information when conducting database searching and de novo sequencing. Here, we use a combination of infrared ion spectroscopy and quantum-chemical calculations to assign the structures of two ETD-generated w-type fragment ions. These ions are spontaneously formed from ETD-generated z• -type fragments by neutral loss of 33 Da in peptides containing a cysteine residue. Analysis of the infrared ion spectra confirms that these z• -ions expel a thiol radical (SH• ) and that a vinyl C═C group is formed at the cleavage site. z• -type fragments containing a Cys residue but not at the cleavage site do not spontaneously expel a thiol radical, but only upon additional collisional activation after ETD.

Water Retention of Calcium-Containing Pectin Studied by Quartz Crystal Microbalance and Infrared Spectroscopy with a Humidity Control System.

To examine differences of water-retention mechanisms between pectins with and without Ca2+, quartz crystal microbalance (QCM) and infrared microspectroscopy combined with a humidity-control system were used to analyze differences in amounts and species of adsorbed water to pectins without and with Ca2+. QCM analysis shows that water contents are ∼2-3 times larger for the pectin film with Ca2+ than that without Ca2+. The difference IR spectra suggest that long, medium, and short H-bond water molecules (free, medium, and bound water) are adsorbed to the pectin film without Ca2+. IR peak shifts of C═O of COOH and C-OH suggest that these water molecules are hydrogen-bonded to C═O and C-OH groups. In addition to these water molecules, bulk water is adsorbed. IR OH band areas fitted by four Gaussian components show that bulk water is mainly adsorbed to the pectin film with Ca2+, possibly among skeletal chains of pectin bridged by Ca2+.

Application of Chemometrics Approaches to Analysis of Mid-Infrared Spectra of Ibuprofen Diluted in Supercritical Carbon Dioxide.

This work represents a comprehensive analysis of mid-infrared (mid-IR) spectra of ibuprofen diluted in supercritical CO2 (in the temperature range of 40-90 ℃ and at the CO2 density corresponding to 1.3 of its critical value). The study employed mathematical approaches based on data matrix analysis such as two-dimensional cross-correlation analysis (2D-COS) and principal component analysis (PCA). Two-dimensional cross-correlation analysis allowed us to reveal correlations between the spectral contributions constituting the analytical spectral band and assigned to certain ibuprofen conformers, as well as the significance of these correlations. It has been shown that the considerable increase in the total intensity of the analytical spectral band, proportional to the equilibrium ibuprofen concentration in the supercritical CO2 phase, is accompanied by certain redistribution of intensities of the spectral components related to the corresponding conformers. The PCA allowed us to determine the changes of intensities of individual spectral contributions for each thermodynamic point in the considered temperature range. It has been shown that these two complementary methods provide more precise information that may be used as the initial data in the classical analysis of spectral data based on spectral curve deconvolution into individual spectral contributions.

Rapid Prediction of Low (<1%) trans Fat Content in Edible Oils and Fast Food Lipid Extracts by Infrared Spectroscopy and Partial Least Squares Regression.

The United States Food and Drug Administration (FDA) ruled that partially hydrogenated oils (PHO), the major dietary source of industrially produced trans fat (TF), were no longer "generally recognized as safe (GRAS)" for any use in human food. Consequently, the objective of this study was to develop a rapid screening procedure using attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy in conjunction with partial least squares regression (PLSR) for the quantitative and accurate prediction of low concentrations of trans fatty acid (TFAs) (<1% of total fatty acids (FAs)). Broad-based calibration models were developed for a combined set of samples consisting of edible oils and fast food lipid extracts. Predicted concentrations of TFAs in the two matrices showed good correlation with the primary reference data generated by gas chromatography (GC) (R2 > 0.99) and high accuracy as evidenced by low root-mean-square error of cross-validation (RMSECV) values. The lowest TFA concentration, determined by GC to be 0.13% of total FAs, was accurately predicted by ATR-FTIR/PLSR as 0.18% of total FAs. This simple, rapid ATR-FTIR/PLSR methodology has the potential for use as a screening alternative to conventional gas chromatographic methods for predicting the TFA content of edible oils and food lipid extracts for regulatory purposes and quality control of raw material and processed food. PRACTICAL APPLICATIONS: FDA ruled that partially hydrogenated oils were no longer "generally recognized as safe (GRAS)" for any use in human food. Consequently, we have proposed a rapid screening procedure, based on infrared spectroscopy and chemometrics, to rapidly and accurately predict low concentrations of trans fatty acids (<1% of total fatty acids) in edible oils and food lipid extracts.

Antioxidant Activity and Infrared Spectroscopy Analysis of Alcoholic Extracts Obtained from Paecilomyces hepiali (Ascomycetes).

This research studies the influence of substrate on the antioxidant activity of alcohol extracts of Paecilomyces hepiali. We used corn, rice, millet, and peas as substrates. Antioxidant activity was measured with the DPPH radical scavenging method. Concentrations of extracts (6.25, 3.12, 1.56, 0.78, and 0.39 mg/mL) were applied in all evaluations. Overall antioxidant activity was expressed as the concentration of substrate that decreased DPPH radical levels by 50% (IC50DPPH) for 7 methanol and 7 ethanol extracts. A comparison of IC50DPPH allowed us to conclude that the methanol extracts are more active in scavenging stable DPPH radicals than are the ethanol extracts. The substrate with antioxidant properties most suitable for cultivation of P. hepiali was rice supplemented with non-defatted soy flour. The extract most effective in scavenging stable radicals was the methanol extract of sample 4 (IC50DPPH = 2.33 mg/mL) cultivated on rice with nondefatted soy flour. The methanol extract of sample 7 cultivated on peas was less effective (IC50DPPH = 11.50 mg/mL). By crystallizing these extracts, we managed to obtain sufficient quantities of 6 samples in a solid state, for which infrared spectra were measured and confirmed the presence of amino acids in the extracts.

Infrared Nanospectroscopy Reveals the Chemical Nature of Pit Membranes in Water-Conducting Cells of the Plant Xylem.

In the xylem of angiosperm plants, microscopic pits through the secondary cell walls connect the water-conducting vessels. Cellulosic meshes originated from primary walls, and middle lamella between adjacent vessels, called the pit membrane, separates one conduit from another. The intricate structure of the nano-sized pores in pit membranes enables the passage of water under negative pressure without hydraulic failure due to obstruction by gas bubbles (i.e. embolism) under normal conditions or mild drought stress. Since the chemical composition of pit membranes affects embolism formation and bubble behavior, we directly measured pit membrane composition in Populus nigra wood. Here, we characterized the chemical composition of cell wall structures by synchrotron infrared nanospectroscopy and atomic force microscopy-infrared nanospectroscopy with high spatial resolution. Characteristic peaks of cellulose, phenolic compounds, and proteins were found in the intervessel pit membranes of P. nigra wood. In addition, the vessel to parenchyma pit membranes and developing cell walls of the vascular cambium showed clear signals of cellulose, proteins, and pectin. We did not find a distinct peak of lignin and other compounds in these structures. Our investigation of the complex chemical composition of intervessel pit membranes furthers our understanding of the flow of water and bubbles between neighboring conduits. The advances presented here pave the way for further label-free studies related to the nanochemistry of plant cell components.

Cyanylated Cysteine Reports Site-Specific Changes at Protein-Protein-Binding Interfaces Without Perturbation.

To investigate the cyanylated cysteine vibrational probe group's ability to report on binding-induced changes along a protein-protein interface, the probe group was incorporated at several sites in a peptide of the calmodulin (CaM)-binding domain of skeletal muscle myosin light chain kinase. Isothermal titration calorimetry was used to determine the binding thermodynamics between calmodulin and each peptide. For all probe positions, the binding affinity was nearly identical to that of the unlabeled peptide. The CN stretching infrared band was collected for each peptide free in solution and bound to calmodulin. Binding-induced shifts in the IR spectral frequencies were correlated with estimated solvent accessibility based on molecular dynamics simulations. This work generally suggests (1) that site-specific incorporation of this vibrational probe group does not cause major perturbations to its local structural environment and (2) that this small probe group might be used quite broadly to map dynamic protein-binding interfaces. However, site-specific perturbations due to artificial labeling groups can be somewhat unpredictable and should be evaluated on a site-by-site basis through complementary measurements. A fully quantitative, simulation-based interpretation of the rich probe IR spectra is still needed but appears to be possible given recent advances in simulation techniques.

An Improved Weighted Partial Least Squares Method Coupled with Near Infrared Spectroscopy for Rapid Determination of Multiple Components and Anti-Oxidant Activity of Pu-Erh Tea.

Background: Pu-erh tea is a unique microbially fermented tea, which distinctive chemical constituents and activities are worthy of systematic study. Near infrared spectroscopy (NIR) coupled with suitable chemometrics approaches can rapidly and accurately quantitatively analyze multiple compounds in samples. Methods: In this study, an improved weighted partial least squares (PLS) algorithm combined with near infrared spectroscopy (NIR) was used to construct a fast calibration model for determining four main components, i.e., tea polyphenols, tea polysaccharide, total flavonoids, theanine content, and further determine the total antioxidant capacity of pu-erh tea. Results: The final correlation coefficients R square for tea polyphenols, tea polysaccharide, total flavonoids content, theanine content, and total antioxidant capacity were 0.8288, 0.8403, 0.8415, 0.8537 and 0.8682, respectively. Conclusions: The current study provided a comprehensive study of four main ingredients and activity of pu-erh tea, and demonstrated that NIR spectroscopy technology coupled with multivariate calibration analysis could be successfully applied to pu-erh tea quality assessment.

Determination of honey adulteration with beet sugar and corn syrup using infrared spectroscopy and genetic-algorithm-based multivariate calibration.

BACKGROUND: Fourier transform infrared spectroscopy (FTIR) equipped with attenuated total reflectance accessory was used to determine honey adulteration. Adulterated honey samples were prepared by adding corn syrup, beet sugar and water as adulterants to the pure honey samples in various amounts. The spectra of adulterated and pure honey samples (n = 209) were recorded between 4000 and 600 cm-1 wavenumber range. RESULTS: Genetic-algorithm-based inverse least squares (GILS) and partial least squares (PLS) methods were used to determine honey content and amount of adulterants. Results indicated that the multivariate calibration generated with GILS could produce successful models with standard error of cross-validation in the range 0.97-2.52%, and standard error of prediction between 0.90 and 2.19% (% w/w) for all the components contained in the adulterated samples. Similar results were obtained with PLS, generating slightly larger standard error of cross-validation and standard error of prediction values. CONCLUSION: The fact that the models were generated with several honey samples coming from various different botanical and geographical origins, quite successful results were obtained for the detection of adulterated honey samples with a simple Fourier transform infrared spectroscopy technique. Having a genetic algorithm for variable selection helped to build somewhat better models with GILS compared with PLS. © 2018 Society of Chemical Industry.