From Bench to Worktop: Rapid Evaluation of Nutritional Parameters in Liquid Foodstuffs by IR Spectroscopy

We evaluated the use of attenuated total reflectance infrared spectroscopy for simultaneous in situ quantification of the nutritional composition of liquid food stuffs in the industrial kitchen context. Different methodologies were compared, including dry and wet acquisition along with instrument parameters and measurement times of 4 and 60 s. The most effective technique was 1-minute measurement, with prediction errors of 2.6, 0.7, 1.0, 2.2, 0.8, 2.4 g/100 mL and 150 Kcal, for carbohydrates, proteins, fat, sugars, saturated fat, water and energy values, respectively.The 4-second method resulted in larger errors but was more applicable for inline measurements. Dry measurements successfully predicted the fractions of proteins, fat, carbohydrates, and sugars, relative to total solids. An app was created to facilitate implementation in a kitchen environment. Compared with other techniques recommended by the FAO, the approach offered a simple alternative for simultaneous prediction of nutritional parameters in an industrial kitchen set-up

Multimodal Vibrational Studies of Drug Uptake In Vitro: Is the Whole Greater than the Sum of Their Parts?

Herein, we investigated the use of multimodal Raman and infrared (IR) spectroscopic microscopy for the elucidation of drug uptake and subsequent cellular responses. Firstly, we compared different methods for the analysis of the combined data. Secondly, we evaluated whether the combined analysis provided enough benefits to justify the fusion of the data. A459 cells inoculated with doxorubicin (DOX) at different times were fixed and analysed using each technique. Raman spectroscopy provided high sensitivity to DOX and enabled an accurate estimation of the drug uptake at each time point, whereas IR provided a better insight into the resultant changes in the biochemical composition of the cell. In terms of benefits of data fusion, 2D correlation analysis allowed the study of the relationship between IR and Raman variables, whereas the joint analysis of IR and Raman enabled the correlation of the different variables to be monitored over time. In summary, the complementary nature of IR and Raman makes the combination of these vibrational techniques an appealing tool to follow drug kinetics and cellular response. Funding information: H2020 Marie Skłodowska-Curie Actions, Grant/Award Number: Spectro-metrics- 020-MSCA-IF-2017 Project ID:79628; National Science Centre, Grant/Award Number: UMO 2016/23/B/NZ4/0137

Quantification and Identification of Microproteinuria using Ultrafiltration and ATR-FTIR Spectroscopy

The presence of low amounts of specific proteins in urine can be an indicator of diagnosis and prognosis of several diseases including renal failure and cancer. Hence, there is an urgent need for Point-of-care (PoC) methods, which can quantify microproteinuria levels (30-300 ppm) and identify the major proteins associated with the microproteinuria. In this study, we coupled ultracentrifugation with attenuated total reflectance-Fourier transform infrared (ATR-FTIR) to identify and quantify proteins in urine at low parts per million levels. The process involves the preconcentration of proteins from 500 μL of urine using an ultrafiltration device. After several washings, the isolated proteins are dried onto the ATR crystal forming a thin film. Imaging studies showed that the absorbance of the protein bands was linear with the amount of mass deposited on the crystal. The methodology was first evaluated with artificial urine spiked with 30-300 ppm of albumin. The calibration showed acceptable linearity (R2 = 0.97) and a limit of detection of 6.7 ppm. Linear relationships were also observed from urine of healthy subjects spiked with microproteinuria concentrations of albumin, immunoglobulin, and hemoglobin, giving a prediction error of the spiked concentration of 23 ppm. When multiple proteins were spiked into the real urine, multivariate analysis was able to decompose the data set into the different proteins, but the multicomponent evaluation was challenging for proteins at low levels. Although the introduction of a preprocessing step reduces the PoC capability of the method, it largely increases its performance, showing great potential as a tool for the diagnosis and prognosis of several illnesses affecting urine proteic compositio

Determining the Age of Spoiled Milk from Dried Films Using Attenuated Reflection Fourier Transform Infrared (ATR FT-IR) Spectroscopy

Milk spoilage is an inevitable occurrence, which generates waste and can result in food poisoning. When milk spoils, the off-flavor and curdling are due to excessive proliferation of various bacteria which causes pH changes. Time, temperature, environment,and previous handling practice all affects the spoilage rate. There is a need for a fast reliable and accurate method that can identify in situearly spoilage of milk. Here we show the ability of attenuated total reflectionFourier transformed infrared spectroscopy(ATR FT-IR) in conjunction with multivariate data analysis to predict the age of milk. We found that dried films vastly increased the absorbance of important biomolecules within milk such as lipids, proteins,and sugars, compared to an unchanged milk sample. This allowed us to note the minor discrepancies that happened in spoilage. Spoilt milk was characterizedby bands associated with increased lipids, proteins, lactic acid; and a decrease in carbohydrates. A semiquantitative prediction model for milk spoilage at room temperature demonstrated ATR FT-IRspectroscopy can predict milk age with a root mean square error of prediction of approximately 14 hours.The model showed poor performance in the first 40 hours but the predictions improved significantly after this time. The experimental procedure proposed for detecting biomolecules within milk has the potential to improve common practice. Furthermore, the model would be a starting point for a newer and improved methods to predict the spoilage date of milk, with potential commercial uses to reduce food waste and costs to the milk industry

Towards the Point of Care and Noninvasive Classification of Bladder Cancer from Urine Sediment Infrared Spectroscopy. Spectral differentiation of normal, abnormal and cancer patients

Bladder cancer (BC) is the 9th cancer cause of death and one of most cost-intensive in the world. The diagnostic tools are still not at all satisfactory. Herein we evaluated the potential of infrared spectroscopy to detect molecular changes that precede and accompany the carcinogenesis in voided urine sediment. We collected 165 samples from patients being diagnosed for BC and measured them with attenuated total reflectance Fourier transformed infrared spectroscopy (ATR FTIR). Samples were primarily divided into three groups according to cytology that indicated the presence of normal, abnormal and cancer cells. ATR FTIR spectra of sediments were analyzed with the use of partial least square discriminant analysis (PLSDA). The 1800–750 cm− 1 region discriminated the three groups with selectivity and sensitivity values around 68% using cytology as a reference method. These cross-validation values (which were found significant according to a permutation test) were comparable to the sensitivity and specificity values of cytology versus the gold standard (histology). The average spectra of each class and the regression vectors of the PLS-DA indicated that an increased content of carbohydrates and nucleic acids as well as transformations of protein secondary structures were the main discriminators of healthy patients from abnormal and cancer groups. Additionally, we revised the obtained classification according to diagnosis made on histopathological assessment of bladder sections. We finally discuss the potential of the technique to be used as a Point of Care (PoC) testing tool

Empirical Study on the Effects of Acquisition Parameters for FTIR Hyperspectral Imaging of Brain Tissue

Fourier transform infrared (FTIR) spectroscopic imaging is a powerful technique for molecular imaging of pathologies associated with the nervous systems including multiple sclerosis research. However, there is no standard methodology or standardized protocol for FTIR imaging of tissue sections that maximize the ability to discriminate between the molecular, white and granular layers, which is essential in the investigation of the mechanism of demyelination process. Tissue sections are heterogeneous, complex and delicate, hence the parameters to generate high quality images in minimal time becomes essential in the modern clinical laboratory. This article presents an FTIR spectroscopic imaging study of post-mortem human brain tissue testing the effects of various measurement parameters and data analysis methods on image quality and acquisition time. Hyperspectral images acquired from the same region of a tissue using a range of the most common optical and collection parameters in different combinations were compared. These included magnification (4× and 15×), number of co-added scans (1, 4, 8, 16, 32, 64 and 128 scans) and spectral resolution (4, 8 and 16 cm−1). Images were compared in terms of acquisition time, signal-to-noise (S/N) ratio, and accuracy of the discrimination between three major tissue types in a section from the cerebellum (white matter, granular and molecular layers). In the latter case, unsupervised k-means cluster (KMC) analysis was employed to generate images from the hyperspectral images, which were compared to a reference image. The classification accuracy for tissue class discrimination was highest for the 4× magnifying objective, with 4 cm−1 spectral resolution and 128 co-added scans

A 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 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, 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 two minutes, 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

Synchrotron Macro ATR-FTIR Microspectroscopy for High Resolution Chemical Mapping of Single Cells

Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy has been used widely for probing the molecular properties of materials. Coupling a synchrotron infrared (IR) beam to an ATR element using a high numerical aperture (NA) microscope objective enhances the spatial resolution, relative to transmission or transflectance microspectroscopy, by a factor proportional to the refractive index (n) of the ATR element. This work presents the development of the synchrotron macro ATR-FTIR microspectroscopy at Australian Synchrotron Infrared Microspectroscopy (IRM) Beamline, and demonstrates that high quality FTIR chemical maps of single cells and tissues can be achieved at an enhanced spatial resolution. The so-called “hybrid” macro ATR-FTIR device was developed by modifying the cantilever arm of a standard Bruker macro ATR-FTIR unit to accept germanium (Ge) ATR elements with different facet sizes (i.e. 1 mm, 250 μm and 100 μm in diameter) suitable for different types of sample surfaces. We demonstrated the capability of the technique for high-resolution single cell analysis of malaria-infected red blood cells, individual neurons in a brain tissue and cellular structures of a Eucalyptus leaf. The ability to measure a range of samples from soft membranes to hard cell wall structures demonstrates the potential of the technique for high-resolution chemical mapping across a broad range of applications in biology, medicine and environmental science

Infrared based saliva screening test for COVID-19

Abstract: Severe acute respiratory syndrome coronavirus 2 has resulted in an unprecedented need for diagnostic testing that is critical in controlling the spread of COVID-19. We propose a portable infrared spectrometer with purpose-built transflection accessory for rapid point-of-care detection of COVID-19 markers in saliva. Initially, purified virion particles were characterized with Raman spectroscopy, synchrotron infrared (IR) and AFM-IR. A data set comprising 171 transflection infrared spectra from 29 patients testing positive for SARS-CoV-2 by RT-qPCR and 28 testing negative, was modeled using Monte Carlo Double Cross Validation with 50 randomized test and model sets. The testing sensitivity was 93 % (27/29) with a specificity of 82 % (23/28) that included positive samples on the limit of detection for RT-qPCR. Here, we demonstrate a proof-of-concept high throughput infrared COVID-19 test that is rapid, inexpensive, portable and utilizes sample self-collection thus minimizing the risk to healthcare workers and ideally suited to mass screening