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Molecular Pharmaceutics Nov 2023Confocal Raman spectroscopy is being assessed as a tool with which to quantify the rate and extent of drug uptake to and its clearance from target sites of action within...
Confocal Raman spectroscopy is being assessed as a tool with which to quantify the rate and extent of drug uptake to and its clearance from target sites of action within the viable epidermis below the skin's stratum corneum (SC) barrier. The objective of this research was to confirm that Raman can interrogate drug disposition within the living layers of the skin (where many topical drugs elicit their pharmacological effects) and to identify procedures by which Raman signal attenuation with increasing skin depth may be corrected and normalized so that metrics descriptive of topical bioavailability may be identified. It was first shown in experiments on skin cross-sections parallel to the skin surface that the amide I signal, originating primarily from keratin, was quite constant with depth into the skin and could be used to correct for signal attenuation when confocal Raman data were acquired in a "top-down" fashion. Then, using 4-cyanophenol (CP) as a model skin penetrant with a strong Raman-active C≡N functionality, a series of uptake and clearance experiments, performed as a function of time, demonstrated clearly that normalized spectroscopic data were able to detect the penetrant to at least 40-80 μm into the skin and to distinguish the disposition of CP from different vehicles. Metrics related to local bioavailability (and potentially bioequivalence) included areas under the normalized C≡N signal versus depth profiles and elimination rate constants deduced post-removal of the formulations. Finally, Raman measurements were made with an approved dermatological drug, crisaborole, for which delivery from a fully saturated formulation into the skin layers just below the SC was detectable.
Topics: Skin Absorption; Spectrum Analysis, Raman; Skin; Epidermis; Biological Availability; Microscopy, Confocal
PubMed: 37801410
DOI: 10.1021/acs.molpharmaceut.3c00755 -
Toxins Oct 2023Mycotoxin contamination on food and feed can have deleterious effect on human and animal health. Agricultural crops may contain one or more mycotoxin compounds;...
Mycotoxin contamination on food and feed can have deleterious effect on human and animal health. Agricultural crops may contain one or more mycotoxin compounds; therefore, a good multiplex detection method is desirable to ensure food safety. In this study, we developed a rapid method using label-free surface-enhanced Raman spectroscopy (SERS) to simultaneously detect three common types of mycotoxins found on corn, namely aflatoxin B1 (AFB1), zearalenone (ZEN), and ochratoxin A (OTA). The intrinsic chemical fingerprint from each mycotoxin was characterized by their unique Raman spectra, enabling clear discrimination between them. The limit of detection (LOD) of AFB1, ZEN, and OTA on corn were 10 ppb (32 nM), 20 ppb (64 nM), and 100 ppb (248 nM), respectively. Multivariate statistical analysis was used to predict concentrations of AFB1, ZEN, and OTA up to 1.5 ppm (4.8 µM) based on the SERS spectra of known concentrations, resulting in a correlation coefficient of 0.74, 0.89, and 0.72, respectively. The sampling time was less than 30 min per sample. The application of label-free SERS and multivariate analysis is a promising method for rapid and simultaneous detection of mycotoxins in corn and may be extended to other types of mycotoxins and crops.
Topics: Animals; Humans; Mycotoxins; Zea mays; Spectrum Analysis, Raman; Zearalenone; Aflatoxin B1; Crops, Agricultural; Food Contamination; Multivariate Analysis; Limit of Detection
PubMed: 37888641
DOI: 10.3390/toxins15100610 -
Journal of Controlled Release :... Dec 2023A correlative methodology for label-free chemical imaging of soft tissue has been developed, combining non-linear optical spectroscopies and mass spectrometry to achieve...
A correlative methodology for label-free chemical imaging of soft tissue has been developed, combining non-linear optical spectroscopies and mass spectrometry to achieve sub-micron spatial resolution and critically improved drug detection sensitivity. The approach was applied to visualise the kinetics of drug reservoir formation within human skin following in vitro topical treatment with a commercial diclofenac gel. Non-destructive optical spectroscopic techniques, namely stimulated Raman scattering, second harmonic generation and two photon fluorescence microscopies, were used to provide chemical and structural contrast. The same tissue sections were subsequently analysed by secondary ion mass spectrometry, which offered higher sensitivity for diclofenac detection throughout the epidermis and dermis. A method was developed to combine the optical and mass spectrometric datasets using image registration techniques. The label-free, high-resolution visualisation of tissue structure coupled with sensitive chemical detection offers a powerful method for drug biodistribution studies in the skin that impact directly on topical pharmaceutical product development.
Topics: Humans; Diclofenac; Tissue Distribution; Skin; Spectrum Analysis, Raman; Mass Spectrometry
PubMed: 37858627
DOI: 10.1016/j.jconrel.2023.10.026 -
Journal of Biomedical Optics Jan 2024Photoacoustic microscopy (PAM) offers advantages in high-resolution and high-contrast imaging of biomedical chromophores. The speed of imaging is critical for leveraging... (Review)
Review
SIGNIFICANCE
Photoacoustic microscopy (PAM) offers advantages in high-resolution and high-contrast imaging of biomedical chromophores. The speed of imaging is critical for leveraging these benefits in both preclinical and clinical settings. Ongoing technological innovations have substantially boosted PAM's imaging speed, enabling real-time monitoring of dynamic biological processes.
AIM
This concise review synthesizes historical context and current advancements in high-speed PAM, with an emphasis on developments enabled by ultrafast lasers, scanning mechanisms, and advanced imaging processing methods.
APPROACH
We examine cutting-edge innovations across multiple facets of PAM, including light sources, scanning and detection systems, and computational techniques and explore their representative applications in biomedical research.
RESULTS
This work delineates the challenges that persist in achieving optimal high-speed PAM performance and forecasts its prospective impact on biomedical imaging.
CONCLUSIONS
Recognizing the current limitations, breaking through the drawbacks, and adopting the optimal combination of each technology will lead to the realization of ultimate high-speed PAM for both fundamental research and clinical translation.
Topics: Microscopy; Prospective Studies; Photoacoustic Techniques; Spectrum Analysis; Lasers
PubMed: 38323297
DOI: 10.1117/1.JBO.29.S1.S11521 -
PloS One 2024In this study, a total of 30 elements (essential and non-essential or toxic) were determined in 25 foods consumed in Italy by children aged 0-6 months and produced in...
In this study, a total of 30 elements (essential and non-essential or toxic) were determined in 25 foods consumed in Italy by children aged 0-6 months and produced in Europe. Inductively Coupled Plasma-Atomic Emission Spectrometry and Inductively Coupled Plasma-Mass Spectrometry were used as measurement techniques for the elements of interest. The estimated intakes for one-year-old infants were compared with risk estimators and nutritional requirements. Data indicate that commercially available baby food in Italy provides an excellent contribution for Mn, Cu, Fe, Zn, Ca, K, and P, covering up to approximately 70% of the adequate intake (AI) for an infant aged 6-12 months. The intake of detectable toxic elements was always below the safety limit: even the most concentrated toxic elements never exceeded about 86% of the Provisional Tolerable Weekly Intake (PTWI). This result indicates that the analyzed baby food is of good quality and does not pose risks to children's health.
Topics: Infant; Child; Humans; Infant Food; Spectrum Analysis; Nutritional Requirements; Italy; Europe
PubMed: 38386635
DOI: 10.1371/journal.pone.0297158 -
Spectrochimica Acta. Part A, Molecular... Dec 2023Due to the background interference from biological samples, detecting viruses using surface-enhanced Raman scattering (SERS) in clinical samples is challenging. This...
Due to the background interference from biological samples, detecting viruses using surface-enhanced Raman scattering (SERS) in clinical samples is challenging. This study is based on SERS by reducing sodium borohydride and aggregating silver nanoparticles to develop suitable virus detection "hot spot." The monkeypox virus and human papillomavirus fingerprints were quickly obtained, tested, and identified in serum and artificial vaginal discharge, respectively, by combining the principal component analysis method. Therefore, these viruses were successfully identified in the biological background. In addition, the lowest detection limit was 100 copies/mL showing good reproducibility and signal-to-noise ratio. The concentration-dependent curve of the monkeypox virus had a good linear relationship. This method helps solve the SERS signal interference problem in complex biological samples, with low detection limits and high selectivity in virus characterization and quantitative analysis. Therefore, this method has a reasonable prospect of clinical application.
Topics: Humans; Spectrum Analysis, Raman; Metal Nanoparticles; Reproducibility of Results; Silver; Viruses; Limit of Detection
PubMed: 37406546
DOI: 10.1016/j.saa.2023.123087 -
Biosensors & Bioelectronics Feb 2024Early diagnosis and accurate assessment of tumor development facilitate early bladder cancer resection and initiation of drug therapy. This study enabled an early,...
Early diagnosis and accurate assessment of tumor development facilitate early bladder cancer resection and initiation of drug therapy. This study enabled an early, accurate, label-free, noninvasive diagnosis of bladder tumors by analyzing nano-biomarkers in a single drop of urine through surface-enhanced Raman spectroscopy (SERS). In a standard N-butyl-N-4-hydroxybutyl nitrosamine-induced rat model of bladder cancer, cancer stage and polyp tumor development were monitored using a small endoscope with a diameter of 1.2 mm in a minimally invasive manner without the need to kill the rats. Samples were divided into cancer-free, early-stage, and polyp-form cancer. Training data were classified according to micro-cystoscopic 5-aminolevulinic acid fluorescence diagnosis, and specimens were postmortem verified through histopathological analysis. A drop of urine from each sample group was placed on an Au-coated zinc oxide nanoporous chip to filter nano-biomaterials and selectively enhance the Raman signals of nanoscale analytes via SERS. Principal component analysis was used to reduce the dimensionality of the collected Raman spectra, and partial least squares discriminant analysis was used to find diagnostic clusters based on the labeled samples. The combination of SERS and machine learning achieved an accuracy ≥99.6% in diagnosing both early- and polyp-stage bladder tumors. With an area under the receiver operating characteristic curve greater than 0.996, the accuracy of the diagnosis in the rat model suggests that SERS-based diagnostic methods are promising when coupled with machine learning. Low-cost, label-free, and noninvasive surface-enhanced Raman spectra are ideal for developing clinically relevant point-of-care diagnostic techniques.
Topics: Rats; Animals; Spectrum Analysis, Raman; Early Detection of Cancer; Biosensing Techniques; Urinary Bladder Neoplasms; Algorithms
PubMed: 38081101
DOI: 10.1016/j.bios.2023.115915 -
Spectrochimica Acta. Part A, Molecular... Oct 2023Monoclonal antibodies provide highly specific and effective therapies for the treatment of chronic diseases. These protein-based therapeutics, or drug substances, are...
Monoclonal antibodies provide highly specific and effective therapies for the treatment of chronic diseases. These protein-based therapeutics, or drug substances, are transported in single used plastic packaging to fill finish sites. According to good manufacturing practice guidelines, each drug substance needs to be identified before manufacturing of the drug product. However, considering their complex structure, it is challenging to correctly identify therapeutic proteins in an efficient manner. Common analytical techniques for therapeutic protein identification are SDS-gel electrophoresis, enzyme linked immunosorbent assays, high performance liquid chromatography and mass spectrometry-based assays. Although effective in correctly identifying the protein therapeutic, most of these techniques need extensive sample preparation and removal of samples from their containers. This step not only risks contamination but the sample taken for the identification is destroyed and cannot be re-used. Moreover, these techniques are often time consuming, sometimes taking several days to process. Here, we address these challenges by developing a rapid and non-destructive identification technique for monoclonal antibody-based drug substances. Raman spectroscopy in combination with chemometrics were used to identify three monoclonal antibody drug substances. This study explored the impact of laser exposure, time out of refrigerator and multiple freeze thaw cycles on the stability of monoclonal antibodies. and demonstrated the potential of using Raman spectroscopy for the identification of protein-based drug substances in the biopharmaceutical industry.
Topics: Spectrum Analysis, Raman; Antibodies, Monoclonal; Mass Spectrometry; Chromatography, High Pressure Liquid
PubMed: 37209478
DOI: 10.1016/j.saa.2023.122872 -
Sensors (Basel, Switzerland) Jul 2023This paper presents a comprehensive study on the development of models and soft sensors required for the implementation of the automated bioreactor feeding of Chinese...
This paper presents a comprehensive study on the development of models and soft sensors required for the implementation of the automated bioreactor feeding of Chinese hamster ovary (CHO) cells using Raman spectroscopy and chemometric methods. This study integrates various methods, such as partial least squares regression and variable importance in projection and competitive adaptive reweighted sampling, and highlights their effectiveness in overcoming challenges such as high dimensionality, multicollinearity and outlier detection in Raman spectra. This paper emphasizes the importance of data preprocessing and the relationship between independent and dependent variables in model construction. It also describes the development of a simulation environment whose core is a model of CHO cell kinetics. The latter allows the development of advanced control algorithms for nutrient dosing and the observation of the effects of different parameters on the growth and productivity of CHO cells. All developed models were validated and demonstrated to have a high robustness and predictive accuracy, which were reflected in a 40% reduction in the root mean square error compared to established methods. The results of this study provide valuable insights into the practical application of these methods in the field of monitoring and automated cell feeding and make an important contribution to the further development of process analytical technology in the bioprocess industry.
Topics: Cricetinae; Animals; Cricetulus; CHO Cells; Bioreactors; Spectrum Analysis, Raman; Least-Squares Analysis
PubMed: 37514911
DOI: 10.3390/s23146618 -
International Journal of Molecular... Aug 2023Circulating cell-free DNA (ccfDNA) is a liquid biopsy biomaterial attracting significant attention for the implementation of precision medicine diagnostics. Deeper...
Circulating cell-free DNA (ccfDNA) is a liquid biopsy biomaterial attracting significant attention for the implementation of precision medicine diagnostics. Deeper knowledge related to its structure and biology would enable the development of such applications. In this study, we employed Raman spectroscopy to unravel the biomolecular profile of human ccfDNA in health and disease. We established reference Raman spectra of ccfDNA samples from healthy males and females with different conditions, including cancer and diabetes, extracting information about their chemical composition. Comparative observations showed a distinct spectral pattern in ccfDNA from breast cancer patients taking neoadjuvant therapy. Raman analysis of ccfDNA from healthy, prediabetic, and diabetic males uncovered some differences in their biomolecular fingerprints. We also studied ccfDNA released from human benign and cancer cell lines and compared it to their respective gDNA, confirming it mirrors its cellular origin. Overall, we explored for the first time Raman spectroscopy in the study of ccfDNA and provided spectra of samples from different sources. Our findings introduce Raman spectroscopy as a new approach to implementing liquid biopsy diagnostics worthy of further elaboration.
Topics: Male; Female; Humans; Spectrum Analysis, Raman; Cell-Free Nucleic Acids; Liquid Biopsy; Breast Neoplasms
PubMed: 37569759
DOI: 10.3390/ijms241512384