Raman Probe Research Articles

Impact of Automation in Pharmaceutical Industry on Roles and Responsibilities of Quality Assurance: A Review

Automation is the use of different control technologies in sectors with minimal or decreased human interference to operate a range of procedures. Automation has made its way to the pharmaceutical industries in the recent past. Automation has been implemented in production, packaging, labeling, and warehousing departments. Production of personalized medicines has become a reality after implementing automated machines. The R and D sector has also been affected by incorporating the latest technologies. The conventional functions of Quality Assurance (QA) department were limited to preparing standard operating procedures (SOPs), carrying out audits, qualification, and validation of equipment and processes. This review focuses on the implementation of automated technologies in pharmaceutical industries and the impact it has had on the pharmaceutical quality assurance department. This article covers topics such as the Raman probe and the different ways it has been made use of in the pharmaceutical industries. The conventional role played by the QA department has also been enlisted while also mentioning how they change with the implementation of automated technologies.

Investigations on the origin of strain variation in the zinc-blende phase along the depth of GaP/Si(1 1 1) using spatially resolved polarized and wavelength dependent Raman spectroscopy

The surface and micro structure variations in the technologically important GaP/Si hetero-structures are investigated by the spatially resolved polarized Raman spectroscopy. The hetero-structures probed from two different directions 〈1 1 1〉 and <11¯0> reveal the presence of strain distributed zinc-blende (ZB) GaP phases and the dominance of wurtzite (WZ) phase near the GaP-Si interface. The azimuthal dependence of the optical phonon intensity of nucleating layer endorses the dominance of WZ phase at the interface. The study illustrates that spatially resolved polarized Raman probe, across the hetero-junction is extremely useful for distinguishing the WZ and ZB stacks. It can positively be utilized for the integration of crystal phase quantum structures of III-V semiconductors on the Si and Ge substrates, without any physical damage to the structures.

Terminal Alkynes as Raman Probes of α-Synuclein in Solution and in Cells.

Conformational changes in α-synuclein (α-syn) are central to its biological function and Parkinson's disease pathology. Here, terminal alkynes (homopropargylglycine) were employed as environmentally sensitive Raman probes at residues 1, 5, 116, and 127 to characterize soluble (disordered), micelle-bound (α-helical), and fibrillar (β-sheet) α-syn. Along with the full-length protein, a disease-related C-terminal truncation (1-115) was also studied. For the first time, β-sheet α-syn amyloid structure was detected by the amide-I band in N27 dopaminergic rat cells, where a reciprocal relationship between levels of fibrils and lipids was seen. Site-specific spectral features of the terminal alkynes also revealed the heterogeneity of the cellular environment. This work shows the versatility of Raman microspectroscopy and the power of unnatural amino acids in providing structural and residue-level insights in solution and in cells.

A highly sensitive SERS quenching nanosensor for the determination of tumor necrosis factor alpha in blood

Tumor necrosis factor alpha (TNF-α) is a cytokine that plays a critical role in medical conditions such cardiovascular diseases, rheumatoid arthritis, inflammatory bowel disease, Alzheimer’s and cancer. Herein, we present a new method for the determination of TNF-α by surface enhanced Raman spectroscopy (SERS). A new benzothiazole azo dye (BAN) was used as a Raman probe to detect the cytokine after its selective extraction from blood plasma using a target-specific antibody-functionalised extractor chip. The disulfide bond structure of the extracted TNF-α was reduced to generate free sulfhydryl (SH), terminal groups that adsorb preferentially onto a BAN-functionalised SERS substrate and displace the BAN Raman reporter on the substrate surface. This causes the SERS spectrum of BAN to quench proportionally with the cytokine concentration. Using this SERS quenching sensor, TNF-α was quantified down to 1 × 10−14 M (173 pg/L). The quantification of the cytokine by the SERS quenching method was cross-validated against enzyme-linked immunosorbent assay (ELISA) and the percent agreement between the two measurements was found to be 93.39 %. Since many proteins and peptides have disulfide bonds in their molecular structures, the new SERS quenching method can be extended for their ultrasensitive quantification after selective extraction from biological fluids.

A Miniature Fibre-Optic Raman Probe Fabricated by Ultrafast Laser-Assisted Etching.

Optical biopsy describes a range of medical procedures in which light is used to investigate disease in the body, often in hard-to-reach regions via optical fibres. Optical biopsies can reveal a multitude of diagnostic information to aid therapeutic diagnosis and treatment with higher specificity and shorter delay than traditional surgical techniques. One specific type of optical biopsy relies on Raman spectroscopy to differentiate tissue types at the molecular level and has been used successfully to stage cancer. However, complex micro-optical systems are usually needed at the distal end to optimise the signal-to-noise properties of the Raman signal collected. Manufacturing these devices, particularly in a way suitable for large scale adoption, remains a critical challenge. In this paper, we describe a novel fibre-fed micro-optic system designed for efficient signal delivery and collection during a Raman spectroscopy-based optical biopsy. Crucially, we fabricate the device using a direct-laser-writing technique known as ultrafast laser-assisted etching which is scalable and allows components to be aligned passively. The Raman probe has a sub-millimetre diameter and offers confocal signal collection with 71.3% ± 1.5% collection efficiency over a 0.8 numerical aperture. Proof of concept spectral measurements were performed on mouse intestinal tissue and compared with results obtained using a commercial Raman microscope.

Flexible paper-based SERS substrate strategy for rapid detection of methyl parathion on the surface of fruit.

Herein, we reported a simple, flexible and sensitive surface-enhanced Raman scattering (SERS) substrate to detect methyl parathion residues in real life. The substrate was fabricated by filter paper and gold nanoparticles (Au NPs) with excellent reproducibility and stability. First, Au NPs were synthesized by the seed mediated growth method and assembled to the filter paper through immersion. The Raman probe molecule 4-MBA was used to evaluate performance of the substrate for an optimized signal using a portable Raman spectrometer coupled with 785nm laser. Then, the paper-based substrate was applied to detect methyl parathion standard solution whose detection limit was down to 0.011μg/cm2, and the linear range was between 0.018μg/cm2 and 0.354μg/cm2. Afterwards, actual sample (apple) spiked with methyl parathion was taken to verify the practicality of the substrate by a simple way of "press-peel off". The recovery rate was ranged from 94.09% to 98.72%, indicating that this method is reliable in actual sample detection without complicated pretreatment steps. This work demonstrates that the flexible paper-based substrate combined with portable Raman instruments can be potentially applied to on-site detection of hazardous substances in the field of food safety.

Phase correlation during two-photon resonance process in an active cavity.

In this paper, we experimentally demonstrate a strong correlation between the frequencies of the Raman pump and the Raman probe inside an optically pumped Raman laser. We show that the correlation is due to rapid adjustment of the phase of the dipoles that produce the Raman gain, following a sudden jump in the phase of the Raman pump. A detailed numerical model validates this interpretation of the phase correlation. The width of the spectrum of the beat between the Raman pump and the Raman laser is significantly narrowed due to this correlation. As a result, the minimum measurable change in the cavity length, for a given linewidth of the Raman pump laser, is substantially reduced. Therefore, this finding is expected to enhance the sensitivity of such a laser in various metrological applications (e.g., accelerometry).

Polydiacetylene-based ultrastrong bioorthogonal Raman probes for targeted live-cell Raman imaging

Live-cell Raman imaging based on bioorthogonal Raman probes with distinct signals in the cellular Raman-silent region (1800–2800 cm−1) has attracted great interest in recent years. We report here a class of water-soluble and biocompatible polydiacetylenes with intrinsic ultrastrong alkyne Raman signals that locate in this region for organelle-targeting live-cell Raman imaging. Using a host-guest topochemical polymerization strategy, we have synthesized a water-soluble and functionalizable master polydiacetylene, namely poly(deca-4,6-diynedioic acid) (PDDA), which possesses significantly enhanced (up to ~104 fold) alkyne vibration compared to conventional alkyne Raman probes. In addition, PDDA can be used as a general platform for multi-functional ultrastrong Raman probes. We achieve high quality live-cell stimulated Raman scattering imaging on the basis of modified PDDA. The polydiacetylene-based Raman probes represent ultrastrong intrinsic Raman imaging agents in the Raman-silent region (without any Raman enhancer), and the flexible functionalization of this material holds great promise for its potential diverse applications.

Focus on photonics, spectrometry, and spectroscopy

Focus on photonics, spectrometry, and spectroscopy

The highly sensitive determination of serotonin by using gold nanoparticles (Au NPs) with a localized surface plasmon resonance (LSPR) absorption wavelength in the visible region.

The development of improved methods for the synthesis of monodisperse gold nanoparticles (Au NPs) is of high priority because they can be used as substrates for surface-enhanced Raman scattering (SERS) applications relating to biological lipids. Herein, Au NPs have been successfully synthesized via a seed-mediated growth method. The LSPR peak is controlled via adjusting the gold nanoseed component, and different fabrication methods were studied to establish the effect of sonication time on NP size. The simple, facile, and room-temperature method is based on a conventional ultrasonic bath, which leads to ultrasonic energy effects on the size and morphology of the Au NPs. This research offers new opportunities for the production of highly monodispersed spherical Au NPs without the use of a magnetic stirrer method, as evidenced by ultraviolet-visible reflectance spectra and scanning electron microscopy (SEM) analysis. SEM images indicate that the spherical Au NP colloidal particles are stable and reliable, which paves the way for their use as a nanostructured biosensor platform that can be exploited for multiple applications, for example, in materials science, sensing, catalysis, medicine, food safety, biomedicine, etc. The highest enhancement factor that could be achieved in terms of the SERS enhancement activity of these Au NP arrays was determined using 10−9 M serotonin (5-hydroxytryptamine, 5-HT) as the Raman probe molecules.

Cellular internalization mechanism of novel Raman probes designed for plant cells.

Diphenylacetylene derivatives containing different polymeric components, poly(l-lysine) (pLys) or tetra(ethylene glycol) (TEG) were designed as novel Raman imaging probes with high Raman sensitivity and low cytotoxicity in living plant cells. The pLys-conjugated probe is internalized via an endocytosis-dependent pathway, whereas TEG-conjugated probe most likely induces direct penetration into the plant cells.

In Situ Monitoring of the "Point Discharge" Induced Antibacterial Process by the Onsite Formation of a Raman Probe.

Electroporation induced by the "point discharge" effect is an effective technique for bacteria inactivation. Rapidly monitoring the electroporation-induced inactivation process is important for screening nanomaterials with high antimicrobial performance. In this study, we develop a facile strategy to in situ monitor the electroporation induced antimicrobial mechanism based on the surface-enhanced Raman scattering (SERS) effect of the Au-nanotip arrays. Owning to the high local-electric field (∼107 V m-1) generated on the Au nanotips, the bacteria are rapidly electroporated and effectively inactivated with ≥99.9% reduction in bacteria colony counts by only applying an external voltage of +0.8 V for 10 s. The related inactivation mechanism is directly verified by the formation of the Prussian blue (PB) nanocrystals by leaking of the uptaken [Fe(CN)6]3- ions from the cleavage area on the cell membrane. These [Fe(CN)6]3- ions react with Fe2+ to form PB nanocrystals onsite as soon as they leak out. The characteristic peak of PB in the cellular Raman-silent region provides a collective monitoring approach for the destruction of microorganisms. The present strategy not only develops a facial method for future use in evaluating electroporation materials, but also paves a rapid way for offering accurate information on some antibacterial and antitumor processes.

Formation of the Majiayao gold deposit, Jiaodong Peninsula, eastern China: Constraints from fluid inclusions, H–O–S–Pb isotopes, and pyrite Rb–Sr age

The Majiayao gold deposit at Qixia in Shandong Province is a typical gold deposit of medium–low temperature hydrothermal veins. This gold deposit is one of the most important types in the Jiaodong Peninsula. Four stages have been identified based on the cross‐cutting relationships and mineralogical and textural characteristics. The study of quartz vein fluid inclusions show that the inclusions can be divided into three categories: pure liquid or pure gas phase inclusions (type S), two‐phase gas and liquid inclusions (type W), and three‐phase inclusions rich in CO2 (type C). The results of laser Raman probe analysis show that the fluid inclusions are generally rich in CO2, thus forming a typical CO2–H2O–NaCl system. The fluid temperature results show that in stage I, only the type W fluid inclusions are identified by microthermometry, the homogenized temperatures (to liquid) are 210–343°C and mostly range from 260 to 280°C, the range of salinity is 1.57–10.98 wt.% NaCl, with a peak value of 8–11 wt.% NaCl, and the density is 0.65–1.02 g/cm3, with a peak value of 0.81–0.87 g/cm3; in stages II and III, the primary fluid inclusion assemblages include both type W and C inclusions, which show similar phase‐transition temperatures and are trapped between 190 and 300°C, and the peak values of salinities are 4–10 wt.% NaCl equivalent; and in stage IV, the total homogenization temperatures (Th‐TOT) into liquid range from 120 to 220°C, and the range of salinities is 4.2–7.9 wt.% NaCl. These values indicate that temperature, salinity, and density gradually decreased from the early to late mineralization stage. The ore‐forming fluid of the deposit was a thermal liquid system with medium–low temperature, medium–low salinity, and low density and was rich in reducing CO2. The H–O isotope shows that the ore‐forming fluids were dominated by metamorphic fluid with only minor amounts of meteoric water involved in the ore‐forming process. According to the S, Pb, and Sr isotopes analysis, it indicated that the source of ore‐forming materials is mainly crustal fluid. The measured Rb–Sr isochron age of pyrite is 123.4 ± 2.9 Ma, which indicates that the metallogenic age corresponded to the Early Cretaceous.

Hydrophobic Paper-Based SERS Sensor Using Gold Nanoparticles Arranged on Graphene Oxide Flakes.

Paper-based surface-enhanced Raman scattering (SERS) sensors have garnered much attention in the past decade owing to their ubiquity, ease of fabrication, and environmentally friendly substrate. The main drawbacks of a paper substrate for a SERS sensor are its high porosity, inherent hygroscopic nature, and hydrophilic surface property, which reduce the sensitivity and reproducibility of the SERS sensor. Here, we propose a simple, quick, convenient, and economical method for hydrophilic to hydrophobic surface modification of paper, while enhancing its mechanical and moisture-resistant properties. The hydrophobic paper (h-paper) was obtained by spin-coating diluted polydimethylsiloxane (PDMS) solution onto the filter paper, resulting in h-paper with an increased contact angle of up to ≈130°. To complete the h-paper-based SERS substrate, gold nanoparticles arranged on graphene oxide (AuNPs@GO) were synthesized using UV photoreduction, followed by drop-casting of AuNPs@GO solution on the h-paper substrate. The enhancement of the SERS signal was then assessed by attaching a rhodamine 6G (R6G) molecule as a Raman probe material to the h-paper-based SERS substrate. The limit of detection was 10 nM with an R2 of 0.966. The presented SERS sensor was also tested to detect a thiram at the micromolar level. We expect that our proposed AuNPs@GO/h-paper-based SERS sensor could be applied to point-of-care diagnostics applications in daily life and in spacecraft.

In situ monitoring of the crystalline state of active pharmaceutical ingredients during high-shear wet granulation using a low-frequency Raman probe

Optimization of manufacturing processes based on scientific evidence is important in the quality control of active pharmaceutical ingredients (APIs) and drug products, particularly when crystal forms change during production, which could affect subsequent drug performance. In this study, we verified crystalline states using various crystal faces and excipients during high-shear wet granulation based on non-contact low-frequency (LF) Raman probe monitoring. Four model drugs [indomethacin (IND), acetaminophen (APAP), theophylline (TP), and caffeine (CAF) polymorphs and cocrystals] were mixed with microcrystalline cellulose and hydroxypropyl cellulose with the addition of water over time. The LF Raman probe showed comparatively high sensitivity in monitoring 5–20% APAP and IND in a wet mass. Notably, as observed from the characteristic LF Raman peak shifts, form I TP and CAF and their cocrystals were more susceptible to transformation to the monohydrate form than form II. This method was also shown to be applicable in monitoring a commercial formulation of eight excipients and revealed crystalline transformations after 15 min of mixing. Therefore, probe-type LF Raman spectroscopy can be successfully employed to distinguish and monitor the crystalline state of APIs in real time during high-shear wet granulation, in which there is a risk of crystal transformation.

Silver Nanocubes Coated in Ceria: Core/Shell Size Effects on Light-Induced Charge Transfer.

Plasmonic sensitization of semiconductors is an attractive approach to increase light-induced photocatalytic performance; one method is to use plasmonic nanostructures in core@shell geometry. The occurrence and mechanism of synergetic effects in photocatalysis of such geometries are under intense debate and proposed to occur either through light-induced charge transfer (CT) or through thermal effects. This study focuses on the relation between the dimensions of Ag@CeO2 nanocubes, the wavelength-dependent efficiency, and the mechanism of light-induced direct CT. A 4-mercaptobenzoic acid (4-MBA) linker between core and shell acts as a Raman probe for CT. For all Ag@CeO2 nanocubes, CT increases with decreasing excitation wavelength, with notable increase at and below 514 nm. This is fully explainable by CT from silver to the 4-MBA LUMO, with the increase for excitation wavelengths that exceed the Ag/4-MBA LUMO gap of 2.28 eV (543 nm). A second general trend observed is an increase in CT yield with ceria shell thickness, which is assigned to relaxation of the excited electron further into the ceria conduction band, potentially producing defects.

Side‐Viewing Endoscopic Raman Spectroscopy for Angle‐Resolved Analysis of Luminal Organs

AbstractThere are many medical applications requiring angle‐resolved analysis of luminal organs. One example is lung cancer detection. Early diagnosis is the most effective way to tackle this disease. This, however, remains a challenge due to the lack of accurate detection technology. The problem is exacerbated in cases of peripheral lung cancer growing on narrow bronchi that are difficult to probe. Here, an endoscopic Raman spectroscopy device is demonstrated with a side‐viewing functionality that enables circumferential scanning spectral measurements inside thin conduits. A tubular micro rotary stepping actuator is custom‐designed and integrated with a Raman probe, for the first time, to scan a probing laser beam sideways for angle‐resolved local Raman analysis with no aid of tissue labeling, toward enabling detection of lesion‐induced biochemical changes in vivo and in real time. A microfabricated prototype is evaluated using test chemicals, harvested animal lung tissue ex vivo, as well as a murine colon model in situ and human skin in vivo. All the test results show excellent agreements with reported reference data while revealing &gt;99% wavenumber accuracies. The study indicates that microactuator‐assisted endoscopic Raman spectroscopy is a promising technology for luminal tissue analysis and encourages further studies of its sensitivity in lesion detection.

High-Frequency Raman Analysis in Biological Tissues Using Dual-Wavelength Excitation Raman Spectroscopy

A dual-wavelength excitation Raman probe with laser inputs at 866 nm or 1064 nm is customized and integrated into a compact Raman spectrometer that is based on an InGaAs detector. Under 1064 nm illumination, the spectrometer detects fingerprint Raman signals below 2000 cm-1. While under 866 nm illumination, the spectral range is extended to cover high-frequency region (2400-4000 cm-1) that includes major C-H and O-H Raman vibrations. We demonstrate that the dual excitation InGaAs Raman is beneficial in detecting high-frequency Raman signals, especially water contents in high-fluorescent biological samples such as human dental tissues, grape skin, and plum skin due to the suppressed fluorescence interference.

Using SERS-based microfluidic paper-based device (μPAD) for calibration-free quantitative measurement of AMI cardiac biomarkers

Recently, surface enhanced Raman scattering (SERS) has attracted much attention in medical diagnosis applications owing to better detection sensitivity and lower limit of detection (LOD) than colorimetric detection. In this paper, a novel calibration-free SERS−based μPAD with multi-reaction zones for simultaneous quantitative detection of multiple cardiac biomarkers — GPBB, CK−MB and cTnT for early diagnosis and prognosis of acute myocardial infarction (AMI) are presented. Three distinct Raman probes were synthesised, subsequently conjugated with respective detecting antibodies and used as SERS nanotags for cardiac biomarker detection. Using a conventional calibration curve, quantitative simultaneous measurement of multiple cardiac biomarkers on SERS-based μPAD was performed based on the characteristic Raman spectral features of each reporter used in different nanotags. However, a calibration free point-of-care testing device is required for fast screening to rule-in and rule-out AMI patients. Partial least squares predictive models were developed and incorporated into the immunosensing system, to accurately quantify the three unknown cardiac biomarkers levels in serum based on the previously obtained Raman spectral data. This method allows absolute quantitative measurement when conventional calibration curve fails to provide accurate estimation of cardiac biomarkers, especially at low and high concentration ranges. Under an optimised condition, the LOD of our SERS-based μPAD was identified at 8, 10, and 1 pg mL−1, for GPBB, CK−MB and cTnT, respectively, which is well below the clinical cutoff values. Therefore, this proof-of-concept technique shows significant potential for highly sensitive quantitative detection of multiplex cardiac biomarkers in human serum to expedite medical decisions for enhanced patient care.

Tailor-made porous striped gold-silver nanowires for surface enhanced Raman scattering based trace detection of β-hydroxybutyric acid

This paper presents a novel fabrication of series of striped (S) and porous striped (PS) gold-silver alloy nanowires (Au-Ag alloy NWs). First, S-Au-Ag alloy NWs were fabricated using anodic aluminum oxide (AAO) template-based electrodeposition technique. The length of the Au-Ag alloy segments was easily tailored by adjusting the deposition pulse time. The surface roughening of Au-Ag alloy nanowires was controlled via dealloying method, resulting in formation of periodic nanogaps between Au alloy nanodisks. This structure was called PS-Au-Ag alloy NWs. The structure of the gap between Au alloy nanodisks was proved to be the key parameter to influence the SERS enhancement ability of nanomaterials. The SERS performance was performed using 4-mercaptobenzoic acid (4-MBA) as a Raman probe molecule. The remarkable SERS enhancement factor of a wire was about 105. The uniformity of NWs was in an acceptable value of relative standard deviation and the NWs showed good stability during 3 months storage time. Moreover, this work was the first demonstration of β-hydroxybutyric acid (BHB) detection using SERS as signal readout with low detection limit about 11 nM. It has been shown that a single PS-Au-Ag alloy NWs exhibited great promise in highly sensitive SERS substrates.