Handheld Raman Spectroscopy Research Articles

Non-Invasive Monitoring of Ethanol and Methanol Levels in Grape-Derived Pisco Distillate by Vibrational Spectroscopy.

Handheld Raman and portable FT-IR spectroscopy devices were evaluated for fast and non-invasive determination of methanol and ethanol levels in Peruvian Pisco. Commercial Peruvian Pisco (n = 171) samples were kindly provided by the UNALM Alliance for Research in Alcohol and its Derivatives (Lima, Peru) and supplemented by purchases at grocery and online stores. Pisco spectra were collected on handheld Raman spectrometers equipped with either a 1064 nm or a 785 nm excitation laser and a portable infrared unit operating in transmission mode. The alcohol levels were determined by GC–MS. Calibration models used partial least-squares regression (PLSR) to develop prediction algorithms. GC–MS data revealed that 10% of Pisco samples had ethanol levels lower than 38%, indicating possible water dilution. Methanol levels ranged from 10 to 130 mg/100 mL, well below the maximum levels allowed for fruit brandies. Handheld Raman equipped with a 1064 nm excitation laser gave the best results for determining ethanol (SEP = 1.2%; RPre = 0.95) and methanol (SEP = 1.8 mg/100 mL; RPre = 0.93). Randomly selected Pisco samples were spiked with methanol (75 to 2800 mg/100 mL), and their Raman spectra were collected through their genuine commercial bottles. The prediction models gave an excellent performance (SEP = 98 mg/100 mL; RPre = 0.97), allowing for the non-destructive and non-contact determination of methanol and ethanol concentrations without opening the bottles.

Raman spectroscopy of fresh feces is a practical tool for identifying animal species and estimating diet characteristics

This study evaluated for the first-time a handheld Raman spectroscopy (RS) device to differentiate animal species and estimate diet characteristics by detecting biomolecules in fresh feces from large (bovine) and small (goat) ruminants, hindgut fermenter (equine), and non-ruminant (swine). The herbivores` diets were forage-based, with access to pasture and ad libitum coastal bermudagrass hay. The pigs received a commercial pelleted grower ration with 160 g/kg of crude protein. Of the 46 fecal samples collected, 21 were from bovine (cattle; 16 cows and 5 calves), 5 from equines (horses, all mares), 8 from goats, and 12 from swine (pigs). Distinct concentrations of undigested biomolecules such as carotenoids, cellulose, and lignin in the feces of each species could be identified by RS. The RS indicated that cattle, goat, and horse diets consisted of 24%, 53%, and 14% fresh grass and 76%, 47%, and 86% hay, respectively. Calves and adult cattle could be distinguished by their feces using the RS. Cellulose, nonstructural carbohydrates, proteins, and fat digestion by ruminants, equines and pigs substantiated RS utilization for nutrition physiology studies in species with different digestive systems. Therefore, RS is a practical tool for identifying animals with different digestive systems and understanding diet specificities.

Identification of Pre-1950 Synthetic Organic Pigments in Artists’ Paints. A Non-Invasive Approach Using Handheld Raman Spectroscopy

There is little information on the actual use of early synthetic organic pigments (SOPs) in art objects, especially those from before 1950. Their presence can, however, pose a challenge to conservation because their chemical composition, as well as their lightfastness and sensitivity to solvents, are often unknown. Here, a study on the non-invasive identification of SOPs in historic pre-1950 varnished paint-outs from artists’ materials manufacturer Royal Talens is presented. The paints were analysed using a handheld Raman device. Spectra were evaluated by recording the spectra of the same samples with a benchtop instrument. This study demonstrated that the identification of SOPs in varnished oil paints with a non-invasive approach is possible and rather straightforward. The handheld Raman device allowed us to identify fourteen SOPs from eight pigment classes. Besides the occurrence of expected and the known SOPs of this time period, there were also some surprising results, like the detection of the triarylcarbonium pigments PG2 and PB8, and the monoazo Mordant Yellow 1.

Non-invasive monitoring of red blood cells during cold storage using handheld Raman spectroscopy.

The current best practices allow for the red blood cells (RBCs) to be stored for prolonged periods in blood banks worldwide. However, due to the individual-related variability in donated blood and RBCs continual degradation within transfusion bags, the quality of stored blood varies considerably. There is currently no method for assessing the blood product quality without compromising the sterility of the unit. This study demonstrates the feasibility of monitoring storage lesion of RBCs in situ while maintaining sterility using an optical approach. A handheld spatially offset Raman spectroscopy (RS) device was employed to non-invasively monitor hemolysis and metabolic changes in 12 red cell concentrate (RCC) units within standard sealed transfusion bags over 7 weeks of cold storage. The donated blood was analyzed in parallel by biochemical (chemical analysis, spectrophotometry, hematology analysis) and RS measurements, which were then correlated through multisource correlation analysis. Raman bands of lactate (857 cm-1 ), glucose (787 cm-1 ), and hemolysis (1003 cm-1 ) were found to correlate strongly with bioanalytical data over the length of storage, with correlation values 0.98 (95% confidence interval [CI]: 0.86-1.00; p=.0001), 0.95 (95% CI: 0.71-0.99; p=.0008) and 0.97 (95% CI: 0.79-1.00; p=.0004) respectively. This study demonstrates the potential of collecting information on the clinical quality of blood units without breaching the sterility using Raman technology. This could significantly benefit quality control of RCC units, patient safety and inventory management in blood banks and hospitals.

Composition analysis of falsified chloroquine phosphate samples seized during the COVID-19 pandemic

The proliferation of falsified medicines can cause serious public health issues, particularly in the context of a global pandemic such as the actual COVID-19 pandemic. Our study involved eight chloroquine phosphate medicines seized in Cameroon, Democratic Republic of Congo and Niger during March and May 2020. These suspect samples were first analyzed in a screening phase using field tools such as handheld Raman spectroscopy (TruScan) and then in a confirmation phase using laboratory tools such as hyperspectral Raman imaging and High Performance Liquid Chromatography (HPLC). The results confirmed the falsified nature of the samples, highlighting the presence of metronidazole at low dose in four samples (16.6, 15.2, 15.2 and 14.5 mg/tab), too low levels of chloroquine in two samples (2.4 and 20.2 mg/tab), and substitution of chloroquine phosphate by paracetamol in one sample (255.7 mg/tab). The results also confirmed that four samples had been adulterated with paracetamol in trace amounts and two of them presented traces of chloramphenicol.

Preliminary Assessment of Parmigiano Reggiano Authenticity by Handheld Raman Spectroscopy.

Raman spectroscopy, and handheld spectrometers in particular, are gaining increasing attention in food quality control as a fast, portable, non-destructive technique. Furthermore, this technology also allows for measuring the intact sample through the packaging and, with respect to near infrared spectroscopy, it is not affected by the water content of the samples. In this work, we evaluate the potential of the methodology to model, by multivariate data analysis, the authenticity of Parmigiano Reggiano cheese, which is one of the most well-known and appreciated hard cheeses worldwide, with protected denomination of origin (PDO). On the other hand, it is also highly subject to counterfeiting. In particular, it is critical to assess the authenticity of grated cheese, to which, under strictly specified conditions, the PDO is extended. To this aim, it would be highly valuable to develop an authenticity model based on a fast, non-destructive technique. In this work, we present preliminary results obtained by a handheld Raman spectrometer and class-modeling (Soft Independent Modeling of Class Analogy, SIMCA), which are extremely promising, showing sensitivity and specificity of 100% for the test set. Moreover, another salient issue, namely the percentage of rind in grated cheese, was addressed by developing a multivariate calibration model based on Raman spectra. It was possible to obtain a prediction error around 5%, with 18% being the maximum content allowed by the production protocol.

On-spot quantification of modafinil in generic medicines purchased from the Internet using handheld Fourier transform-infrared, near-infrared and Raman spectroscopy

Poor quality medicines represent an expanding global public health threat facilitated by the Internet. A recent survey showed that one in five students have used modafinil to enhance learning ability mainly purchased from Internet sources. The aim of this work was to develop on-the-spot and simple methods for the quantification of modafinil in generic medicines using Fourier transform-infrared (FTIR), near-infrared (NIR) and Raman spectroscopy along with partial least square regression (PLSR). Modafinil tablets were measured in intact form using NIR and Raman and in powdered form using FTIR, NIR and Raman. Additionally, powder mixtures of crushed modafinil tablets and excipient(s) were prepared either by diluting the crushed tablets with excipient(s), or sequentially adding excipient(s) to the crushed tablets. Three PLSR models were constructed in MATLAB 2014a from powder mixtures and two from intact and powdered tablets. For FTIR and Raman spectroscopy, PLSR models based on tablets gave linear calibration curve with correlation coefficient (r2) values above 0.94 and a root mean square error of calibration (RMSEC) below 0.96% m/m. Conversely, the PLSR model based on powder sequential addition gave the highest accuracy using the NIR spectra (r2 = 0.99, RMSEC = 1.15% m/m). The latter model showed accuracy in predicting the concentration of the active pharmaceutical ingredient in modafinil generic medicines proving their authenticity. The overall results showed that the combination of the three spectroscopic methods with PLSR offered a rapid technique for authenticating generic modafinil medicines.

Rapid Detection of Cypermethrin by Using Surface-Enhanced Raman Scattering Technique

Cypermethrin is a toxic pesticide in the pyrethroid group. A Surface Enhanced Raman Scattering (SERS) based sensor has been developed to achieve simple pesticide sensing. In this work, rapid detection of cypermethrin by using the handheld Raman spectroscopy coupled with SERS substrate was demonstrated. SERS-active silver nanorods substrate was used to enhance Raman signals of test samples. The effect of exposure time and drop volume of sample was studied for cypermethrin measurement. The results found that the silver nanorods substrate can be used to measure cypermethrin in the range of 10-6 to 10-3 M with a handheld Raman spectrometer. Furthermore, the Raman signal of cypermethrin was confirmed by measuring solid cypermethrin with the standard Raman spectrometer. SERS substrate was competent to detect cypermethrin with a limit of detection (LOD) of 10-6 M.

High-Throughput Phenotyping Approach for Screening Major Carotenoids of Tomato by Handheld Raman Spectroscopy Using Chemometric Methods.

Our objective was to develop a rapid technique for the non-invasive profiling and quantification of major tomato carotenoids using handheld Raman spectroscopy combined with pattern recognition techniques. A total of 106 samples with varying carotenoid profiles were provided by the Ohio State University Tomato Breeding and Genetics program and Lipman Family Farms (Naples, FL, USA). Non-destructive measurement from the surface of tomatoes was performed by a handheld Raman spectrometer equipped with a 1064 nm excitation laser, and data analysis was performed using soft independent modelling of class analogy (SIMCA)), artificial neural network (ANN), and partial least squares regression (PLSR) for classification and quantification purposes. High-performance liquid chromatography (HPLC) and UV/visible spectrophotometry were used for profiling and quantification of major carotenoids. Seven groups were identified based on their carotenoid profile, and supervised classification by SIMCA and ANN clustered samples with 93% and 100% accuracy based on a validation test data, respectively. All-trans-lycopene and β-carotene levels were measured with a UV-visible spectrophotometer, and prediction models were developed using PLSR and ANN. Regression models developed with Raman spectra provided excellent prediction performance by ANN (rpre = 0.9, SEP = 1.1 mg/100 g) and PLSR (rpre = 0.87, SEP = 2.4 mg/100 g) for non-invasive determination of all-trans-lycopene in fruits. Although the number of samples were limited for β-carotene quantification, PLSR modeling showed promising results (rcv = 0.99, SECV = 0.28 mg/100 g). Non-destructive evaluation of tomato carotenoids can be useful for tomato breeders as a simple and rapid tool for developing new varieties with novel profiles and for separating orange varieties with distinct carotenoids (high in β-carotene and high in cis-lycopene).

Non-invasive diagnostics of Liberibacter disease on tomatoes using a hand-held Raman spectrometer.

Hand-held Raman spectroscopy can be used for confirmatory, non-invasive and non-destructive detection and identification of two haplotypes of Liberibacter disease on tomatoes. Using this spectroscopic approach, structural changes in carotenoids, xylan, cellulose and pectin that are associ-ated with this bacterial disease can be determined. 'Candidatus Liberibacter solanacearum' (Lso) is a phloem-limited Gram-negative bacterium that infects crops worldwide. In North America, two haplotypes of Lso (LsoA and LsoB) are transmitted by the potato psyllid, Bactericera cockerelli (Sulč), and infect many solanaceous crops such as potato and tomato. Infected plants exhibit chlorosis, severe stunting, leaf cupping, and scorching. Polymerase chain reaction (PCR) and potato tuber frying are commonly used methods for diagnostics of the plant disease caused by Lso. However, they are time-consuming, costly, destructive to the sample, and often not sensitive enough to detect the pathogen in the early infection stage. Raman spectroscopy (RS) is a noninvasive, nondestructive, analytical technique, which probes chemical composition of analyzed samples. In this study, we demonstrate that Lso infection can be diagnosed by non-invasive spectroscopic analysis of tomato leaves three weeks following infection, before the development of aerial symptoms. In combination with chemometric analyses, Raman spectroscopy allows for 80% accurate diagnostics of Liberibacter disease caused by each of the two different haplotypes. This diagnostics approach is portable and sample agnostic, suggesting that it could be utilized for other crops and could be conducted autonomously.

Handheld confocal Raman spectroscopy (CRS) for objective assessment of skin barrier function and stratification of severity in atopic dermatitis (AD) patients

BackgroundWe developed the first-of-its-kind handheld confocal Raman spectroscopy (CRS) system to quantify the concentration of natural moisturizing factors in the skin. ObjectiveTo evaluate the feasibility of our handheld CRS system and propose a novel quantitative index to measure skin barrier function. MethodsThis prospective study included 30 atopic dermatitis (AD) patients and 14 healthy volunteers. All AD participants were assessed using the Scoring Atopic Dermatitis (SCORAD) severity instrument, a vapometer for trans-epidermal water loss and a moisture meter for skin surface moisture. A handheld CRS operating at 785 nm laser was used to measure the biochemical constituents of the skin up to a depth of ∼100 μm. We trained a linear kernel-based support vector machine (SVM) model for eczema classification based on the water, ceramide and urocanic acid content. A novel Eczema Biochemical Index (EBI) was then formulated using the skin constituents measured from the AD participants to stage disease severity. ResultsThe SVM model used to classify healthy participants and AD patients obtained high cross-validated area under the curve of 0.857 and accuracy of 0.841, with high sensitivity and specificity values of 0.857 and 0.833 respectively. EBI can be used to stratify AD patients of varying severity, based on the biochemical constituents in the skin. ConclusionAs compared to the standard CRS system, the handheld CRS offers higher portability and provides Raman measurements at various body regions with similar sensitivity. This suggests that a handheld CRS device could be a valuable point-of-care resource in both research and clinical use.

First Use of Handheld Raman Spectroscopy to Analyze Omega-3 Fatty Acids in Intact Fish Oil Capsules.

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are commercially important omega-3 fatty acids found in fish oils. Here we demonstrate that a handheld Raman spectrometer can be used to quantitate these compounds in intact fish oil capsules, avoiding oxidizing risk. Partial least squares regression models were prepared by relating Raman spectral variance to EPA and DHA concentrations determined using gas chromatography-mass spectrometry (GC-MS) analysis of fatty acid methyl esters in 15 commercial samples containing 145-473 mg g-1 EPA and 101-260 mg·g-1 DHA. Handheld Fourier transform (FT)-Raman models had root mean square errors of cross-validation of 38 mg g-1, 24 mg g-1, and 32 mg·g-1 for EPA, DHA, and EPA+DHA, respectively. Models generated from a benchtop FT-Raman spectrometer had corresponding errors of 32 mg·g-1, 22 mg·g-1, and 26 mg·g-1. By comparison, average standard deviations from triplicate GC-MS analyses were 11 mg·g-1 for EPA and 9 mg·g-1 for DHA.

Falsified vardenafil tablets available online

It is often reported that falsified medicines have harmful effects on patients both Japan and abroad. In this study, we purchased vardenafil tablets on the internet and investigated their quality and authenticity using visual observations, authenticity investigations, non-destructive tests (handheld NIR and Raman spectroscopy), and quality analyses (active ingredient content and tablet dissolution rate). We used genuine 20-mg Levitra tablets that were sold in Japan and tablets from Bayer AG (Germany) as controls. In April 2015, we obtained 28 samples from 15 websites on the internet. Our authenticity investigations revealed that 11 (40%) were genuine products and 17 (60%) were falsified products. Handheld NIR and Raman results revealed that the falsified products had different spectra to the genuine products. Principal component analysis of the NIR and Raman spectra showed variation among the falsified products. The 11 genuine products were of good quality, and the 17 falsified products were of poor quality. The falsified products contained sildenafil (the active ingredient of Viagra) or tadalafil (the active ingredient of Cialis) instead of vardenafil. Our results show that falsified Vardenafil tablets are sold on the internet and that it is important to prevent illegal internet sales and increase consumer awareness of the presence of falsified medicines.

Quantitation of active pharmaceutical ingredient through the packaging using Raman handheld spectrophotometers: A comparison study

Handheld Raman spectroscopy is actually booming. Recent devices improvements aim at addressing the usual Raman spectroscopy issues: fluorescence with shifted‐excitation Raman difference spectroscopy (SERDS), poor sensitivity with surface enhanced Raman scattering (SERS) and information only about the sample surface with spatially offset Raman spectroscopy (SORS). While qualitative performances of handheld devices are generally well established, the quantitative analysis of pharmaceutical samples remains challenging.The aim of this study was to compare the quantitative performances of three commercially available handheld Raman spectroscopy devices. Two of them (TruScan and IDRaman mini) are equipped with a 785 nm laser wavelength and operate in a conventional backscattering mode. The IDRaman has the Orbital Raster Scanning (ORS) option to increase the analyzed surface. The third device (Resolve) operates with an 830 nm laser wavelength both in backscattering and in SORS modes.The comparative study was carried out on ibuprofen-mannitol-microcrystalline cellulose ternary mixtures. The concentration of ibuprofen ranged from 24 to 52% (w/w) while the proportions of the two excipients were varied to avoid cross-correlation as much as possible. Analyses were performed either directly through a glass vial or with the glass vial in an opaque polypropylene flask, using a validated FT-NIR spectroscopy method as a reference method. Chemometric analyses were carried out with the Partial Least Squares Regression (PLS-R) algorithm. The quantitative models were validated using the total error approach and the ICH Q2 (R1) guidelines with ± 15% as acceptance limits.

Composition of Amphiboles in the Tremolite–Ferro–Actinolite Series by Raman Spectroscopy

Amphiboles are an important family of rock forming minerals, whose identification is crucial in provenance studies as well as in many other fields of geology, archaeology and environmental sciences. This study is aimed to find a quick way to characterize Ca-amphiboles in the tremolite (Ca2Mg5Si8O22(OH)2)–ferro–actinolite (Ca2Fe5Si8O22(OH)2) series. Raman spectroscopy is established as technique to perform non-destructive and quick analysis, with micrometric resolution, able to give the composition in terms of Mg/(Mg + Fe2+) ratio. To exploit the method, a preliminary characterization is performed by Scanning Electron Microscopy coupled with Energy-dispersed X-ray Spectroscopy (SEM-EDS). Two independent methods to evaluate the composition from the Raman data (aiming to an accuracy of about 5%), using the low-wavenumbers part of the spectrum and the OH stretching bands, are developed. The application of the proposed method to micro-Raman mappings and the possible use of handheld Raman spectroscopy to have compositional information on Ca-amphiboles are discussed.

Raman spectroscopy as an early detection tool for rose rosette infection.

Hand-held Raman spectroscopy is a potential tool for a confirmatory, non-invasive, and non-destructive detection and identification of rose rosette disease. Using this spectroscopic approach, structural changes in roses that are associated with this viral infection can be revealed. The commercial rose shrub industry in the United States is one of the largest of its kind. All commercial rose varieties are susceptible to rose rosette disease (RRD), a deadly viral disease vectored by eriophyid mites. This disease is typically diagnosed visually and/or by PCR-based detection assays. The present work demonstrates that Raman spectroscopy can detect RRD in intact leaf tissue. It is shown that chemometric analysis can distinguish between spectra collected from symptomatic and asymptomatic tissue, as well as between healthy and asymptomatic tissue. This method will be useful as an initial screen for RRD prior to PCR analysis to help conserve reagents and save time.

Rapid Discrimination of Malaria- and Dengue-Infected Patients Sera Using Raman Spectroscopy.

Malaria and dengue have overlapping clinical symptoms and are prevalent in the same geographic region (tropical and subtropical), hence precise diagnosis is challenging. The high mortality rate associated with both malaria and dengue could be attributed to "false", "delayed", or "missed" diagnosis. The present study thus aims to stratify malaria and dengue using Raman spectroscopy (RS). In total, 130 human sera were analyzed for model development and double-blinded testing. Principal components linear discriminant analysis (PC-LDA) of acquired RS-spectra could classify malaria and dengue with a minor overlap of 16.7%. Receiver operating characteristic (ROC) analysis of test samples showed sensitivity/specificity of 0.9529 for malaria vs healthy controls (HC) and 0.9584 for dengue vs HC. The Raman findings were complemented by mass spectroscopy (MS)-based metabolite analysis of 8 individuals, each from malaria, dengue, and HC. Several of the metabolites, including amino acids, cell-free DNA, creatinine, and bilirubin, assigned for the predominant RS-bands were also identified by MS and showed similar trends. Our data clearly indicates that RS-based serum analysis using a microprobe has immense potential for early, accurate, and automated detection and discrimination of malaria and dengue, and in the future, it could be extrapolated in field-settings combined with hand-held RS. Further, this approach might be extended to diagnose other closely related infections with similar clinical manifestations.

Enabling Robust and Rapid Raw Material Identification and Release by Handheld Raman Spectroscopy.

A fast, reproducible, non-destructive method to confirm raw material identification in real-time upon material receipt within a warehouse environment is desired. Current practices in pharmaceutical manufacturing often employ compendia methods for raw material identification tests, which require sample preparation prior to time-consuming chemical analysis and often employ subjective spectral comparisons. We have developed, qualified, and validated a rapid objective identity method ("Rapid ID") by Raman spectroscopy using the Bruker BRAVO handheld Raman spectrometer for 46 common raw materials used in upstream and downstream biopharmaceutical cell culture-based processes. Materials in the Raman identification library include amino acids and other solid neat organic chemicals, liquid organics, polyatomic salts, polymers, emulsifiers, peptides, aqueous solutions, and buffers. Selection of reference spectra and hit quality index limit(s) was based upon a comprehensive spectral survey across multiple suppliers and lots to account for normal cause spectral variation. Method repeatability and reproducibility, selectivity, and robustness against various operational and environmental factors (e.g., instrumental variance, material packaging, and thermal effects) were evaluated. Benefits of a handheld Raman Rapid ID approach include significant reduction of the time for raw material quality release from weeks to minutes, enhanced objectivity, and robust data integrity via autonomous electronic reporting. In addition, routine collection of rich spectroscopic data on raw materials can be leveraged to support further continuous improvement initiatives, including routine monitoring of method performance, continuous improvement of the library, proactive detection of shifts in raw material properties, and provision of data for investigations focused on raw materials. Rapid ID methods are consistent with the move toward the principles of Pharma 4.0-high automated processes with continuous process verification and a holistic control strategy.LAY ABSTRACT: A fast, reproducible, non-destructive method is desired to confirm raw material identification in real time upon receipt within a warehouse environment. We have developed, qualified and validated a rapid objective identity method ("Rapid ID") by Raman spectroscopy using the Bruker BRAVO handheld Raman spectrometer for 46 common raw materials used in upstream and downstream biopharmaceutical cell culture-based processes. Benefits of a handheld Raman Rapid ID approach include significant time reduction of raw material quality release from weeks to minutes, enhanced objectivity, and robust data integrity via autonomous electronic reporting. Rapid ID methods are consistent with the move toward the principles of Pharma 4.0: high automated processes with continuous process verification and a holistic control strategy.

Rapid through-container detection of fake spirits and methanol quantification with handheld Raman spectroscopy.

The spirits drinks industry is of significant global economic importance and a major employer worldwide, and the ability to ensure product authenticity and maintain consumer confidence in these high-value products is absolutely essential. Spirit drinks counterfeiting is a worldwide problem, with counterfeiting and adulteration of spirit drinks taking many forms, such as substitution, stretching with lower-grade products, or creation of counterfeits with industrial, surrogate, or locally produced alcohols. Methanol for example, which has been used as a substitute alcohol for ethanol, has a high toxicity in humans. The counterfeiting of spirit drinks is consequently one of the few leading reported types of food fraud which can be directly and unequivocally linked to food safety and health concerns. Here, for the first time, we use handheld Raman spectroscopy with excitation in the near IR (1064 nm) for the through-container differentiation of multiple spirit drinks, detection of multiple chemical markers of counterfeit alcohol, and for the quantification of methanol. We established the limits of detection (LOD) of methanol in the analysed samples from four different spirit types (between 0.23-0.39%), which were considerably lower than a quoted maximum tolerable concentration (MTC) of 2% (v/v) methanol for humans in a 40% alcohol by volume (ABV) spirit drink, and even lower than the general EU limit for naturally occurring methanol in fruit spirits of 0.5% v/v (10 g methanol per L ethanol). We believe that Raman spectroscopy has considerable practicable potential for the rapid in situ through-container detection of counterfeit spirits drinks, as well as for the analysis and protection of other beverages and liquid samples.

Abstract B063: Towards the use of Raman spectroscopy to guide prostate biopsy procedure

Abstract For the first time, molecular information deduced from Raman spectroscopy measurements was spatially correlated with histopathologic analysis to detect and characterize prostate cancer (PCa) using Raman spectroscopy (RS). Ex vivo post-prostatectomy specimen were interrogated using a handheld RS probe. Up to 50 areas (of 500µm in diameter each) per prostate slice were correlated spatially with hematoxylin, phloxine, and saffron (HPS) stained images for detailed histopathologic characteristics, including percentage of benign glands and cancer grade group (International Society of Urological Pathology, ISUP): (1) each measured area was identified with a spot of Indian ink; (2) a photograph was taken at the end of the measurements to build a mask including the prostate slice contour and circles corresponding to the measured areas; (3) the whole prostate slice was reconstructed from the HPS scans guided by anatomic landmarks to insure spatial correspondence with the fresh prostate slice photograph. The Raman spectra were analyzed in terms of univariate statistics of specific Raman peaks corresponding to biomolecules such as DNA/RNA, amino acids, or lipids. In addition, the spectra were submitted to multivariate statistical classification using machine learning algorithms (artificial neural network). In order to test the validity of ex vivo conditions regarding intraoperative (i.e., in vivo) applications, we measured and compared different rat organs (including brain, prostate, ovarian, spleen, kidney, and liver) in in vivo and ex vivo conditions. Previous studies about the use of RS for PCa detection were done exclusively on cell lines or snap-frozen human tissue by considering the overall patient diagnosis. PCa tissues are highly heterogeneous morphologically: cancer glands are intermingled with benign glands and tumors are often multiclonal. Therefore, local histopathologic criteria must be considered. We designed a method that allows us to correlate spatially each Raman spectrum with precise diagnostic criteria and we were able to distinguish extraprostatic from prostatic tissue with an accuracy of 82% (sensibility of 82%, specificity of 83%), malignant from benign prostatic tissue with an accuracy over 86% (sensibility of 87%, specificity of 86%), and different cancer grade groups, for instance, with an accuracy of 84% for distinguishing ISUP grade group 1 from 5. When comparing in vivo with ex vivo rat tissues, preliminary results showed that the difference between average spectra is of the same magnitude than the variability within the same tissue type (same organ from different rats), therefore meaning that ex vivo RS data could be used to train classifier for in vivo diagnosis. Thanks to our robust methodology spatially correlating the optical measurements with local histopathologic characteristics, our work on ex vivo human prostates has proved that we have the capability to characterize the heterogeneity of prostate tissue in the context of PCa. In addition, we established that the use of ex vivo specimen to mimic intraoperative condition is valid. These are promising results, especially in the context of optical guided biopsy procedure to help collect targeted tissue to decrease the false-negative rate nowadays associated with the procedure (~30%). Citation Format: Kelly Aubertin, Mirela Birlea, Michael Pinto, Karl St-Arnaud, Vincent Quoc Trinh, Andrée-Anne Grosset, Frédéric Leblond, Dominique Trudel. Towards the use of Raman spectroscopy to guide prostate biopsy procedure [abstract]. In: Proceedings of the AACR Special Conference: Prostate Cancer: Advances in Basic, Translational, and Clinical Research; 2017 Dec 2-5; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(16 Suppl):Abstract nr B063.