A highly sensitive SERS substrate of porous membrane-Ag NPs for kidney cancer detection.

BACKGROUND: The substrate employed in surface-enhanced Raman spectroscopy (SERS) constitutes an essential element in the cancer detection methodology. In this research, we introduce a three-dimensional (3D) structured SERS substrate that integrates a porous membrane with silver nanoparticles to enhance SERS spectral signals through the utilization of the aggregation effect of silver nanoparticles. This enhancement is crucial because accurate detection results strongly depend on the intensity of specific peaks in Raman spectroscopy. A highly sensitive SERS substrate can significantly improve the accuracy of detection results. RESULTS: We collected 66 plasma samples from individuals with kidney cancer and control individuals, including both bladder cancer patients and healthy individuals. Then, we utilized substrates with and without porous membranes to acquire the SERS spectra of the samples, enabling us to evaluate the enhancement effect of our SERS substrate. The spectral analysis demonstrated enhanced peak intensities in the experimental group (with porous substrate) compared to the control group (without porous substrate). The uniformity and reproducibility of the SERS substrate are also significantly enhanced, which is very helpful for improving the accuracy of detection results. Additionally, the Principal Component Analysis-Linear Discriminant Analysis algorithm (PCA-LDA) was employed to classify the SERS spectra of both groups. In the experimental group, the classification accuracy was 98.5 % for kidney cancer, and 83.3 % for kidney and bladder cancer. Compared to the control group, it improved by 3 % and 12.6 % respectively. SIGNIFICANT: This indicates that our 3D structured SERS substrate combined with multivariate statistical algorithms PCA-LDA can not only improve the accuracy of SERS detection technology in single cancer detection, but also has great potential in multiple cancer detection. This 3D structured SERS substrate is expected to become a new auxiliary means for cancer detection.

Identifying the origin of Yemeni green coffee beans using near infrared spectroscopy: a promising tool for traceability and sustainability.

Yemeni smallholder coffee farmers face several challenges, including the ongoing civil conflict, limited rainfall levels for irrigation, and a lack of post-harvest processing infrastructure. Decades of political instability have affected the quality, accessibility, and reputation of Yemeni coffee beans. Despite these challenges, Yemeni coffee is highly valued for its unique flavor profile and is considered one of the most valuable coffees in the world. Due to its exclusive nature and perceived value, it is also a prime target for food fraud and adulteration. This is the first study to identify the potential of Near Infrared Spectroscopy and chemometrics-more specifically, the discriminant analysis (PCA-LDA)-as a promising, fast, and cost-effective tool for the traceability of Yemeni coffee and sustainability of the Yemeni coffee sector. The NIR spectral signatures of whole green coffee beans from Yemeni regions (n = 124; Al Mahwit, Dhamar, Ibb, Sa'dah, and Sana'a) and other origins (n = 97) were discriminated with accuracy, sensitivity, and specificity ≥ 98% using PCA-LDA models. These results show that the chemical composition of green coffee and other factors captured on the spectral signatures can influence the discrimination of the geographical origin, a crucial component of coffee valuation in the international markets.

Comparison of classification methods performance for defining the best reuse of waste wood material using NIR spectroscopy.

Recycling of post-consumer waste wood material is becoming an increasingly appealing alternative to disposal. However, its huge heterogeneity is calling for an assessment of the material characteristics in order to define the best recycling option and intended reuse. In fact, waste wood comes into a variety of uses/types of wood, along with several levels of contamination, and it can be divided into different categories based on its composition and quality grade. This study provides the measurement of more than a hundred waste wood samples and their characterisation using a hand-held NIR spectrophotometer. Three classification methods, i.e. K-nearest Neighbours (KNN), Principal Component Analysis - Linear Discriminant Analysis (PCA-LDA) and PCA-KNN, have been compared to develop models for the sorting of waste wood in quality categories according to the best-suited reuse. In addition, the classification performance has been investigated as a function of the number of the spectral measurements of the sample and as the average of the spectral measurements. The results showed that PCA-KNN performs better than the other classification methods, especially when the material is ground to 5 cm of particle size and the spectral measurements are averaged across replicates (classification accuracy: 90.9 %). NIR spectroscopy, coupled with chemometrics, turned out to be a promising tool for the real-time sorting of waste wood material, ensuring a more accurate and sustainable waste wood management. Obtaining real-time information about the quality and characteristics of waste wood material translates into a decision of the best recycling option, increasing its recycling potential.

Authentication and identification of Lamiaceae family with cyclic voltammetry fingerprint-PCA-LDA and determination of the used phenolic contents for classification using chromatographic analyses.

Todays, it is essential to evaluate and check the quality of herbal medicines in to protect the public health. As medicinal plants, the extracts of labiate herbs are used directly or indirectly to treat a variety of diseases. Increase in their consumption has led to the fraud in herbal medicines. Hence, modern accurate diagnostic methods must be introduced to differentiate and authenticate these samples. Electrochemical fingerprints have not been evaluated for their capacity to distinguish and classify various genera within a family. Since it is essential to classify, identify, and distinguish between these closely related plants in order to guarantee the quality of the raw materials, the authenticity and quality of 48 dried and fresh Lamiaceae samples, which include Mint, Thyme, Oregano, Satureja, Basil, and Lavender with various geographic origins, were examined. The present study focused on (a) classification and authentication Labiate herbs extracts and (b) identification of active compounds in samples by Gas chromatography and HPLC methods. This was accomplished using principal component analysis (PCA) and PCA-linear discriminate analysis (PCA-LDA). The results of the clustering revealed that PCA-LDA categorized mint species more accurately than PCA. In addition to certain flavonoids including ferulic acid, apigenin, luteolin, and quercetin, HPLC and GC analysis of the ethanolic extract revealed the presence of phenolic acids such as rosmarenic acid, methyl rosmarenate, caffeic acid, cinnamic acid, and chlorogenic acid. Comparing results of PCA-LDA with chromatographic analysis show that the authentication and detection of fraud samples were correctly performed using chemometyrics technique based on CV fingerprints. Even, there was no need to completely identify components of the mint samples.

Rapid discrimination of Brucellosis in sheep using serum Fourier transform infrared spectroscopy combined with PCA-LDA algorithm.

Brucellosis in sheep is an infectious disease caused by Brucella melitensis in sheep. The current conventional serological methods for screening Brucella-infected sheep have the disadvantage of time consuming and low accuracy, so a simple, rapid and highly accurate screening method is needed. The aim of this study was to evaluate the feasibility of diagnosing Brucella-infected sheep by serum samples based on the Fourier transform infrared (FTIR) spectroscopy. In this study, FTIR spectroscopy of serum from Brucella-infected sheep (n = 102) and healthy sheep (n = 125) revealed abnormal protein and lipid metabolism in serum from Brucella-infected sheep compared to healthy sheep. Principal component analysis-Linear discriminant analysis (PCA-LDA) method was used to differentiate the FTIR spectra of serum from Brucella-infected sheep and healthy sheep in the protein band (3700-3090 cm-1) and lipid band (3000-2800 cm-1), and its overall diagnostic accuracy was 100% (sensitivity 100%, specificity 100%). In conclusion, our results suggest that serum FTIR spectroscopy combined with PCA-LDA algorithm has great potential for brucellosis in sheep screening.

Use of ATR-FTIR spectroscopy to differentiate between cirrhotic/non-cirrhotic HCV patients.

BACKGROUND: Conventional techniques to diagnose (HCV) and assess non-cirrhotic/cirrhotic status of the patient for appropriate treatment regime are expensive and invasive. Present available diagnostic tests are expensive as they include multiple screening steps. Therefore, there is a need of cost-effective, less time consuming and minimally invasive alternative diagnostic approaches can be used for effective screening. We propose that (ATR-FTIR) in conjunction with (PCA-LDA),(PCA-QDA) and (SVM) multivariate algorithms can be used as a sensitive tool for detection of HCV infection and to assess non-cirrhotic/cirrhotic status of patients. METHODS: We used 105 sera samples, of which, 55 were from healthy and 50 were from HCV positive individuals. These 50 HCV positive patients were further classified into cirrhotic and non-cirrhotic categories using serum markers and imaging techniques. These samples were freeze dried prior to spectral acquisition then multivariate data classification algorithms were employed to classify these sample types. RESULTS: PCA-LDA and SVM model computed the diagnostic accuracy of 100% for detection of HCV infection. To further classify the non-cirrhotic/cirrhotic status of a patient, diagnostic accuracy of 90.91% for PCA-QDA and 100% for SVM was observed. Internal and external validation for SVM based classifications observed 100% sensitivity and specificity. The confusion matrix generated by PCA-LDA model computed the validation and calibration accuracy showed 100% sensitivity and specificity, by using 2 PCs for HCV infected and healthy individuals. However, when the PCA QDA analysis was done to classify the non-cirrhotic sera samples from cirrhotic sera samples the diagnostic accuracy achieved was 90.91% based on 7 PC's. SVM was also employed for classification and developed model showed the best results with 100% sensitivity and specificity when external validation was applied. CONCLUSIONS: This study provides an initial insight that ATR-FTIR spectroscopy in conjugation with multivariate data classification tools holds a potentialnot onlytoeffectively diagnosis HCV infection but also to assess non-cirrhotic/cirrhotic status of patients.

Label-free surface-enhanced Raman spectroscopy of serum with machine-learning algorithms for gallbladder cancer diagnosis.

Gallbladder cancer (GBC) is a rare but frequently fatal biliary tract malignancy that is typically discovered when it is already advanced. In this study, we investigated a novel technique for the quick and non-invasive diagnosis of GBC based on serum surface-enhanced Raman spectroscopy (SERS). SERS spectra of serum from 41 patients with GBC and 72 normal subjects were recorded. Principal component analysis-linear discriminant analysis (PCA-LDA), and PCA-support vector machine (PCA-SVM), Linear SVM and Gaussian radial basis function-SVM (RBF-SVM) algorithms were used to establish the classification models, respectively. When the Linear SVM was used, the overall diagnostic accuracy for classifying the two groups could achieve 97.1%, and when RBF-SVM was used, the diagnostic sensitivity of GBC was 100%. The results demonstrated that SERS combination with a machine learning algorithm is a promising candidate to be one of the diagnostic tools for GBC in the future.

ATR-FTIR spectrum analysis of plasma samples for rapid identification of recovered COVID-19 individuals.

The development of fast, cheap and reliable methods to determine seroconversion against infectious agents is of great practical importance. In the context of the COVID-19 pandemic, an important issue is to study the rate of formation of the immune layer in the population of different regions, as well as the study of the formation of post-vaccination immunity in individuals after vaccination. Currently, the main method for this kind of research is enzyme immunoassay (ELISA, enzyme-linked immunosorbent assay). This technique is sufficiently sensitive and specific, but it requires significant time and material costs. We investigated the applicability of attenuated total reflection (ATR) Fourier transform infrared (FTIR) spectroscopy associated with machine learning in blood plasma to detect seroconversion against SARS-CoV-2. The study included samples of 60 patients. Clear spectral differences in plasma samples from recovered COVID-19 patients and conditionally healthy donors were identified using multivariate and statistical analysis. The results showed that ATR-FTIR spectroscopy, combined with principal components analysis (PCA) and linear discriminant analysis (LDA) or artificial neural network (ANN), made it possible to efficiently identify specimens from recovered COVID-19 patients. We built classification models based on PCA associated with LDA and ANN. Our analysis led to 87% accuracy for PCA-LDA model and 91% accuracy for ANN, respectively. Based on this proof-of-concept study, we believe this method could offer a simple, label-free, cost-effective tool for detecting seroconversion against SARS-CoV-2. This approach could be used as an alternative to ELISA.

Identify the Virus-like Models for COVID-19 as Bio-Threats: Combining Phage Display, Spectral Detection and Algorithms Analysis.

The rapid identification and recognition of COVID-19 have been challenging since its outbreak. Multiple methods were developed to realize fast monitoring early to prevent and control the pandemic. In addition, it is difficult and unrealistic to apply the actual virus to study and research because of the highly infectious and pathogenic SARS-CoV-2. In this study, the virus-like models were designed and produced to replace the original virus as bio-threats. Three-dimensional excitation-emission matrix fluorescence and Raman spectroscopy were employed for differentiation and recognition among the produced bio-threats and other viruses, proteins, and bacteria. Combined with PCA and LDA analysis, the identification of the models for SARS-CoV-2 was achieved, reaching a correction of 88.9% and 96.3% after cross-validation, respectively. This idea might provide a possible pattern for detecting and controlling SARS-CoV-2 from the perspective of combining optics and algorithms, which could be applied in the early-warning system against COVID-19 or other bio-threats in the future.

Label-Free Characterization of Macrophage Polarization Using Raman Spectroscopy.

Macrophages are important cells of the innate immune system that play many different roles in host defense, a fact that is reflected by their polarization into many distinct subtypes. Depending on their function and phenotype, macrophages can be grossly classified into classically activated macrophages (pro-inflammatory M1 cells), alternatively activated macrophages (anti-inflammatory M2 cells), and non-activated cells (resting M0 cells). A fast, label-free and non-destructive characterization of macrophage phenotypes could be of importance for studying the contribution of the various subtypes to numerous pathologies. In this work, single cell Raman spectroscopic imaging was applied to visualize the characteristic phenotype as well as to discriminate between different human macrophage phenotypes without any label and in a non-destructive manner. Macrophages were derived by differentiation of peripheral blood monocytes of human healthy donors and differently treated to yield M0, M1 and M2 phenotypes, as confirmed by marker analysis using flow cytometry and fluorescence imaging. Raman images of chemically fixed cells of those three macrophage phenotypes were processed using chemometric methods of unmixing (N-FINDR) and discrimination (PCA-LDA). The discrimination models were validated using leave-one donor-out cross-validation. The results show that Raman imaging is able to discriminate between pro- and anti-inflammatory macrophage phenotypes with high accuracy in a non-invasive, non-destructive and label-free manner. The spectral differences observed can be explained by the biochemical characteristics of the different phenotypes.

Label-Free SERS Analysis of Serum Using Ag NPs/Cellulose Nanocrystal/Graphene Oxide Nanocomposite Film Substrate in Screening Colon Cancer.

Label-free surface-enhanced Raman scattering (SERS) analysis shows tremendous potential for the early diagnosis and screening of colon cancer, owing to the advantage of being noninvasive and sensitive. As a clinical diagnostic tool, however, the reproducibility of analytical methods is a priority. Herein, we successfully fabricated Ag NPs/cellulose nanocrystals/graphene oxide (Ag NPs/CNC/GO) nanocomposite film as a uniform SERS active substrate for label-free SERS analysis of clinical serum. The Ag NPs/CNC/GO suspensions by self-assembling GO into CNC solution through in-situ reduction method. Furthermore, we spin-coated the prepared suspensions on the bacterial cellulose membrane (BCM) to form Ag NPs/CNC/GO nanocomposite film. The nanofilm showed excellent sensitivity (LOD = 30 nM) and uniformity (RSD = 14.2%) for Nile Blue A detection. With a proof-of-concept demonstration for the label-free analysis of serum, the nanofilm combined with the principal component analysis-linear discriminant analysis (PCA-LDA) model can be effectively employed for colon cancer screening. The results showed that our model had an overall prediction accuracy of 84.1% for colon cancer (n = 28) and the normal (n = 28), and the specificity and sensitivity were 89.3% and 71.4%, respectively. This study indicated that label-free serum SERS analysis based on Ag NPs/CNC/GO nanocomposite film combined with machine learning holds promise for the early diagnosis of colon cancer.

Heritage and modern wheat varieties discrimination by volatiles profiling. Is it a matter of flavor?

Nowadays, the interest in ancient wheat is increasing and this trend point towards the local production of crops and is connected to sustainability. In this study, two ancient wheat (Solina and Cappelli) and four modern (common and durum) varieties were cultivated in experimental fields sited at three different altitudes for three consecutive years in the Abruzzo region. The six wheat varieties were analysed by solid phase microextraction coupled with gas chromatography-mass spectrometry and a chemometric approach. 149 compounds, most of which are odor active, were identified in 109 wheat samples. Heritage wheat varieties showed a volatile organic compounds (VOCs) profile different from modern varieties along with a characteristic set of odor types. An 82% of correct classification was achieved for heritage wheat varieties. VOCs with floral and herbal odors were the most important odor scents for Solina classification, whereas waxy odor was the most important for Cappelli discrimination.

The Authentication of Gayo Arabica Green Coffee Beans with Different Cherry Processing Methods Using Portable LED-Based Fluorescence Spectroscopy and Chemometrics Analysis.

Aceh is an important region for the production of high-quality Gayo arabica coffee in Indonesia. In this area, several coffee cherry processing methods are well implemented including the honey process (HP), wine process (WP), and natural process (NP). The most significant difference between the three coffee cherry processing methods is the fermentation process: HP is a process of pulped coffee bean fermentation, WP is coffee cherry fermentation, and NP is no fermentation. It is well known that the WP green coffee beans are better in quality and are sold at higher prices compared with the HP and NP green coffee beans. In this present study, we evaluated the utilization of fluorescence information to discriminate Gayo arabica green coffee beans from different cherry processing methods using portable fluorescence spectroscopy and chemometrics analysis. A total of 300 samples were used (n = 100 for HP, WP, and NP, respectively). Each sample consisted of three selected non-defective green coffee beans. Fluorescence spectral data from 348.5 nm to 866.5 nm were obtained by exciting the intact green coffee beans using a portable spectrometer equipped with four 365 nm LED lamps. The result showed that the fermented green coffee beans (HP and WP) were closely mapped and mostly clustered on the left side of PC1, with negative scores. The non-fermented (NP) green coffee beans were clustered mostly on the right of PC1 with positive scores. The results of the classification using partial least squares-discriminant analysis (PLS-DA), linear discriminant analysis (LDA), and principal component analysis-linear discriminant analysis (PCA-LDA) are acceptable, with an accuracy of more than 80% reported. The highest accuracy of prediction of 96.67% was obtained by using the PCA-LDA model. Our recent results show the potential application of portable fluorescence spectroscopy using LED lamps to classify and authenticate the Gayo arabica green coffee beans according to their different cherry processing methods. This innovative method is more affordable and could be easy to implement (in terms of both affordability and practicability) in the coffee industry in Indonesia.

Exploring the potential of paper-based analytical sensors for tea geographical origin authentication.

Tea (Camellia sinensis (L.) Kuntze-surname of German origin) is a popular beverage consumed worldwide due to its health benefits. Its quality depends on measuring features that may discriminate teas from distinct provenances. Protected designation of origin (PDO) is therefore a very useful label for tea quality evaluation. In the present work, antioxidant activity profiles obtained from microfluidic paper-based analytical devices (µPADs) were analyzed by chemometrics to determine the tea geographic origin. Based on the existing literature, we constructed a database containing chemical data from 26 samples and evaluated it by principal component analysis (PCA) coupled to linear discriminant analysis (LDA). Antioxidant activity was an effective LDA predictor for sample discrimination accomplishing accuracies from 75 to 82%. Modeling performance was favored by an external validation method. The best classification model was found using the first nine PCs as input variables. Training samples achieved a perfect success rate, while the test ones were predicted with 83% specificity, 100% sensitivity, and 90% overall accuracy. The modeling robustness was verified by integrating AUC (0.943) from ROC curve. The PCA-LDA approach taken here demonstrated that the teas coming from different countries can be correctly authenticated through µPADs, thus contributing to certificate samples PDO. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05440-1.

Discriminant Analysis PCA-LDA Assisted Surface-Enhanced Raman Spectroscopy for Direct Identification of Malaria-Infected Red Blood Cells.

Various methods for detecting malaria have been developed in recent years, each with its own set of advantages. These methods include microscopic, antigen-based, and molecular-based analysis of blood samples. This study aimed to develop a new, alternative procedure for clinical use by using a large data set of surface-enhanced Raman spectra to distinguish normal and infected red blood cells. PCA-LDA algorithms were used to produce models for separating P. falciparum (3D7)-infected red blood cells and normal red blood cells based on their Raman spectra. Both average normalized spectra and spectral imaging were considered. However, these initial spectra could hardly differentiate normal cells from the infected cells. Then, discrimination analysis was applied to assist in the classification and visualization of the different spectral data sets. The results showed a clear separation in the PCA-LDA coordinate. A blind test was also carried out to evaluate the efficiency of the PCA-LDA separation model and achieved a prediction accuracy of up to 80%. Considering that the PCA-LDA separation accuracy will improve when a larger set of training data is incorporated into the existing database, the proposed method could be highly effective for the identification of malaria-infected red blood cells.

Near-infrared spectroscopy combined with chemometrics to classify cosmetic foundations from a crime scene.

Cosmetic smears are a form of trace evidence that can link the crime scene, suspects, and victims. Foundation and lipstick are the most common sources of cosmetics that can easily smear, with most current research focused on the evidential analysis of lipsticks. This research aims to create a database of cosmetic foundations on different materials and to access the robustness of using Near-infrared with chemometrics as a non-destructive technique to identify unknown samples collected from a crime scene. Small amounts of six shades of three brands of foundations were smeared on clothing materials, which were then analysed with a combination of Near-infrared with chemometric analysis. Principle component analysis (PCA) was used to reduce data dimensionality and explore potential patterns in sample separation and Linear Discriminant Analysis (LDA) was utilised to assign unknown samples to one of the established classes. The selected techniques proved to be promising for database construction and as a preliminary method of analysis, with 93% of the spectra being correctly classified. Notably, darker foundation shades were less likely to be correctly classified (90% classified correctly) compared to lighter ones (96.7% classified correctly). This could not be improved with Standard Normal Variate (SNV) data pre-treatment or selecting specific NIR regions. This finding is of particular importance; according to the Crime Survey for England and Wales (year ending March 2020) police recorded sexual offences demonstrated that those in Mixed and Black or Black British ethnic groups were significantly more likely to be a victim of sexual assault compared to White, Asian or Other ethnic groups. It is, therefore, crucial to add a wide range of foundation shades, particularly of darker tones, to the future database.

Label-free detection of bladder cancer and kidney cancer plasma based on SERS and multivariate statistical algorithm.

In this study, we mainly aimed to investigate the diagnostic potential of surface-enhanced Raman spectroscopy for bladder cancer and kidney cancer which are the most common cancers of the urinary system, and evaluate the classification ability of three statistical algorithms: principal component analysis-linear discriminate analysis (PCA-LDA), partial least square-random forest (PLS-RF), and partial least square-support vector machine (PLS-SVM). The plasma of 26 bladder cancer patients, 38 kidney cancer patients and 39 normal subjects was mixed with the same volume of silver nanoparticles, respectively, and then high-quality SERS signal was obtained. The SERS spectra in the range of 400-1800 cm-1 were compared and analyzed. There were some significant differences in SERS peak intensity, which may reflect the changes in the content of some biomacromolecules in the plasma of cancer patients. Based on the three algorithms of PCA-LDA, PLS-RF and PLS-SVM, the classification accuracy of SERS spectra of plasma from cancer patients and normal subjects was 98.1%, 100% and 100%, respectively. In addition, the classification accuracy of the three diagnostic algorithms to classify the SERS spectra of bladder cancer and kidney cancer was 81.3%, 91.7%, and 98.4%, respectively. This exploratory work demonstrates that SERS combined with PLS-SVM algorithm has superior performance for clinical screening of bladder cancer and kidney cancer through peripheral plasma.

Revealing the Effect of Heat Treatment on the Spectral Pattern of Unifloral Honeys Using Aquaphotomics.

In this study we aimed to investigate the effect of heat treatment on the spectral pattern of honey using near infrared spectroscopy (NIRS). For the research, sunflower, bastard indigo, and acacia honeys were collected from entrusted beekeepers. The honeys were not subject to any treatment before. Samples were treated at 40 °C, 60 °C, 80 °C, and 100 °C for 60, 120, 180, and 240 min. This resulted in 17 levels, including the untreated control samples. The 5-hydroxymethylfurfural (HMF) content of the honeys was determined using the Winkler method. NIRS spectra were recorded using a handheld instrument. Data analysis was performed using ANOVA for the HMF content and multivariate analysis for the NIRS data. For the latter, PCA, PCA-LDA, and PLSR models were built (using the 1300-1600 nm spectral range) and the wavelengths presenting the greatest change induced by the perturbations of temperature and time intervals were collected systematically, based on the difference spectra and the weights of the models. The most contributing wavelengths were used to visualize the spectral pattern changes on the aquagrams in the specific water matrix coordinates. Our results showed that the heat treatment highly contributed to the formation of free or less bonded water, however, the changes in the spectral pattern highly depended on the crystallization phase and the honey type.

Characterization and identification of microplastics using Raman spectroscopy coupled with multivariate analysis.

The manufacture and use of plastic products have resulted in the release and spread of a massive amount of microplastics. Identifying and quantifying microplastics is challenging due to their small size and complicated composition. Although vibrational spectroscopy has been applied to analyze microplastics, its reliability and throughput are limited by the challenges to distinguish the pending alterations manually and the lack of a spectra-based automated microplastic classification model. The present study applied Raman spectroscopy coupled with multivariate analysis to develop a new and robust analytical method to comprehensively interrogate the spectral profiles of seven microplastic references and real microplastic samples post-exposure to environmental stresses. Besides identifying unique Raman peaks of individual microplastics, their whole spectra were separated by principal component analysis (PCA) and linear discriminant analysis (LDA). Support vector machine (SVM) classification achieved an accuracy rate of over 98% for polypropylene, polyethylene terephthalate, polyvinyl chloride, polycarbonate, polyamide, and over 70% for high-density polyethylene and low-density polyethylene. Real microplastic samples from the breakdown of snack boxes, mineral water bottles, juice bottles, and medicine vials were also matched to their chemical components by SVM with an overall sensitivity, specificity, and accuracy of 98.1%, 99.4%, and 99.1%, respectively. Additionally, post-exposure to environmental stressors, 1D PCA-LDA score plots could still distinguish microplastic type, and the developed SVM classification achieved an accuracy of 96.75% in the real-world scenario. These findings prove Raman spectroscopy coupled with multivariate analysis as an ideal tool to distinguish the types and environmental exposure of microplastics, demonstrating great potential for microplastic automatic detection.

NIRS and Aquaphotomics Trace Robusta-to-Arabica Ratio in Liquid Coffee Blends.

Coffee is both a vastly consumed beverage and a chemically complex matrix. For a long time, an arduous chemical analysis was necessary to resolve coffee authentication issues. Despite their demonstrated efficacy, such techniques tend to rely on reference methods or resort to elaborate extraction steps. Near infrared spectroscopy (NIRS) and the aquaphotomics approach, on the other hand, reportedly offer a rapid, reliable, and holistic compositional overview of varying analytes but with little focus on low concentration mixtures of Robusta-to-Arabica coffee. Our study aimed for a comparative assessment of ground coffee adulteration using NIRS and liquid coffee adulteration using the aquaphotomics approach. The aim was to demonstrate the potential of monitoring ground and liquid coffee quality as they are commercially the most available coffee forms. Chemometrics spectra analysis proved capable of distinguishing between the studied samples and efficiently estimating the added Robusta concentrations. An accuracy of 100% was obtained for the varietal discrimination of pure Arabica and Robusta, both in ground and liquid form. Robusta-to-Arabica ratio was predicted with R2CV values of 0.99 and 0.9 in ground and liquid form respectively. Aquagrams results accentuated the peculiarities of the two coffee varieties and their respective blends by designating different water conformations depending on the coffee variety and assigning a particular water absorption spectral pattern (WASP) depending on the blending ratio. Marked spectral features attributed to high hydrogen bonded water characterized Arabica-rich coffee, while those with the higher Robusta content showed an abundance of free water structures. Collectively, the obtained results ascertain the adequacy of NIRS and aquaphotomics as promising alternative tools for the authentication of liquid coffee that can correlate the water-related fingerprint to the Robusta-to-Arabica ratio.