Advanced Pharma Magazine

Edition No. 1

ADVANCED PHARMA Combined SEM and AFM Particle Analysis of Vitamin C Drug Polymorph Structure Analysis With TEM 3D Electron Diffraction Tomography

CASE STUDY: Next Level of Pharmaceutical Inspections

SURFACE-SENSITIVE CHARACTERISATION OF ATOMIC LAYER DEPOSITION FILMS ON DRUG PARTICLES

From Counterfeit to Compliance: Hyperspectral Imaging Secures Drug Quality CASE STUDY: Measurement of Powder or Lyocake Substances Made Easier

MicroCT of Pharmaceutical Packaging

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“ At Quantum Design UK and Ireland, we’re

passionate about providing high-tech scientific instrumentation to researchers, universities, and industries across the UK and Ireland.

Based in Leatherhead, Surrey, we’re a small but dedicated team, bringing deep expertise and a personal touch to everything we do.

We specialise in materials characterisation, cryogenics, non-destructive testing, imaging cameras, microscopy, and spectroscopy. But more than just supplying instruments, we work closely with our customers to understand their unique research challenges and help them find the best solutions.

What sets us apart?

We’re knowledgeable, approachable, and always happy to help. Our Sales and Service departments are second-tonone, offering expert support, training, and advice to make sure our customers get the most out of their equipment.

At Quantum Design UK and Ireland, we take pride in being more than just a supplier—we’re a trusted partner in scientific discovery.”

Dr. Luke Nicholls Acting Managing Director Quantum Design UK and Ireland

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About Us For more than 40 years Quantum Design (QD) has been providing technology solutions to researchers in the fields of physics, chemistry, biotechnology, materials science, and nanotechnology.

Established in 1982 in San Diego, California, Quantum Design is the leading commercial source for automated materials characterisation systems offering a variety of measurement capabilities.

QD instruments are found in the world’s leading research institutions and have become the reference standard for a variety of magnetic and physical property measurements.

Quantum Design instruments are cited in, and provide the data for, more scientific publications than any other instrument in the fields of magnetics and materials characterisation.

An essential part of providing scientific solutions to researchers around the globe is to also offer state-of-the-art instruments from other leading manufacturers.

These manufacturers are chosen by QD not only for their innovative products, but also because they believe in the same level of customer satisfaction and support that scientists have come to expect from QD products worldwide.

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Contents 3-

Foreword

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From Counterfeit to Compliant: Hyperspectral Imaging Secures Drug Quality

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CASE STUDY: High Speed Analysis of Powder and Tablets with Hyperspectral Imagery

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Precession Electron Diffraction for Pharma

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CASE STUDY: Revealing Crystal Structure of Carbamazepine with 3D Electron Diffraction Tomography

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CASE STUDY: Measurement of Powder or Lyocake Substances Made Easier Even in Glass Vials

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Combined SEM and AFM Particle Analysis of Vitamin C with FusionScope

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How PiFM Outperfroms Conventional Techniques

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The Highest Resolution Nanoscale Chemical Analysis Instruments

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MicroCT of Pharmaceuticals and Pharmaceutical Packaging

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CASE STUDY: Next Level of Pharmaceutical Inspections

FOREWORD The UK’s pharmaceutical and life sciences industry is one of the country’s largest and most innovative sectors, underpinning both economic growth and global healthcare leadership. With continued investment in advanced manufacturing, nanotechnology, and drug formulation, researchers and process engineers are placing greater emphasis on materials characterisation to understand surface properties, structural features, and nanoscale chemical interactions.

Quantum Design UK and Ireland (QDUKI), working in partnership with leading international manufacturers, provides advanced analytical and microscopy tools that support pharmaceutical development, manufacturing, and quality assurance.

Two recent examples highlight how these technologies are being applied across the UK to address complex challenges in pharmaceutical research and production.

Dr. Shayz Ikram TECHNICAL DIRECTOR QUANTUM DESIGN UK AND IRELAND

FROM COUNTERFEIT TO COMPLIANCE: HYPERSPECTRAL IMAGING SECURES DRUG QUALITY

Learn more about Hyperspectral Imaging cameras by Specim

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Pharmaceutical products are visually often very similar but can have different chemical compositions or proportions of active pharmaceutical ingredients (API).

Hyperspectral imaging provides a reliable, rapid, and non-invasive way to analyse the chemical composition of drug formulations, such as tablets and capsules, during production The chemical analysis allows: Qualitatively to identify pharmaceutical products with different active ingredients.

Quantitatively to identify pharmaceutical products with different dosages and to measure the uniformity of the active ingredient distribution

Hyperspectral imaging enables reliable monitoring of the pharmaceutical manufacturing processes in real time. It can cover the entire material stream in-line with close to 100% accuracy. This information can help manufacturers ensure their products’ quality, purity, and consistency and improve the manufacturing processes.

Benefits

Figure 1: Hyperspectral imaging can identify products with similar looks but with different active ingredient or concentration in real-time with 100% accuracy

Non-destructive, rapid, and reliable inspection with close to 100% accuracy Inspect 100% of the product stream in real-time Measure the presence, amount, and distribution of API Avoid mix-ups and improve product quality and safety

Case Study

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High-speed analysis of powder and tablets with hyperspectral imaging A common application of NIR spectroscopy is the identification of the active compound or active pharmaceutical ingredient (API) present in a tablet or drug.

A French company, Indatech has developed a new technology for powder and tablet analysis that relies on the innovative use of optical probes using several optical fibres to measure the surface of the sample. All the fibres are connected to a Specim NIR hyperspectral imaging system.

About IndaTech

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Indatech was found in 2009 by Sylvie Roussel and Fabien Chauchard. Two main reasons were that Sylvie had found out that single spectra couldn’t provide the critical information that the customer needed while Fabien had been developing a solution, based on a multipoint imaging with promising results. This convinced them that hyperspectral imaging would be the best in-line technology in the future.

Currently Indatech provides solutions mainly for the pharma, biotech and chemical industry. Their core business is the high-speed inspection of a tablet (hardness, API, homogeneity) and powder either in manufacturing or in process development phase.

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Challenging approach meets hyperspectral imaging The challenge in their approach is the need to measure multiple optical fibres at the same time as well as they must be able to change the configuration of the measurement. Before hyperspectral, we tested standard spectrometer connected to an optical multiplexer but in the end, the measurement took too long,” says Fabien...

So, we decided to try the “hyperspectral imaging and we found out that the pushbroom imaging is the best approach because the spectra are measured in a single shot without any moving parts and the resolution can also be adapted.”

“We started discussing with another manufacturer before we met Mr Timo Hyvärinen at a Pharma Congress in the United States. He presented me the company, as well as the technology behind and I’ve been completely convinced of the performance and the quality ever since.

Specim is very reactive and always tries to find a solution in case of a specific need.

The products are robust since the company has acquired a long experience.

The products have the little things that make the difference in terms of performance and quality.” says Fabien.

Real partnership is a foundation for excellent results “Our solution is now compatible with manufacturing machine speeds, we have been able to inspect from 150 000 to 800 000 tablets per hour based on homogeneity and API concentration.

Specim’s reactivity, quality and flexibility both on delivering the camera and with technical support has met our expectations. We have been able to build a real partnership where our engineers can easily communicate with Specim engineers. This has resulted in continuous improvement in the use of the technology. We have modified the design of our solution based on hyperspectral camera in order to be able to work on different problematics using the same detector such as mixing of powder and fluid, compaction of the tablet, capsule filling, lyophilisation and so on.”

To discuss the Specim range of hyperspectral imaging cameras, get in touch with our Sales Manager, Dr. Luke Nicholls by email luke@qd-uki.co.uk or call 01372 378822.

Webinar

How to use limited electron diffraction data to solve metastable pharmaceutical structure at ambient temperature In this webinar, Dr. Grahame Woollam from Novartis AG, Switzerland and Dr. Partha Das from NanoMEGAS SPRL, Belgium present: “Structural analysis of metastable pharmaceutical loratadine form II, by 3D electron diffraction and DFT+D energy minimisation” based on their paper published in CrystEngComm

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Precession Electron Diffraction

FOR PHARMA

Applications:

Orientation & Phase mapping Strain Mapping Precession enhanced 3D-ED for crystal structure determination Electron Pair Distribution Function (ePDF) for amorphous materials characterisation Electron Pair Distribution Function (ePDF) mapping Electric Field mapping Enhanced EELS and EDX Spectroscopy

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CASE STUDY Revealing Crystal Structure of Carbamazepine with 3D Electron Diffraction Tomography

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For industrial applications, it is of great interest to understand and determine the crystal structure, identify / characterise several possible polymorphs and to detect / quantify the crystallinity / amorphous phase of final products, since many

Carbamazepine (CBZ), is a drug used primarily in the treatment of epilepsy and neuropathic pain. It may be used in schizophrenia along with other medications and as a second line agent in bipolar disorder.

CBZ exists in several polymorphic forms.

important chemical and physical properties depend on the crystal structure properties.

Electron Diffraction Tomography analysis with TEM using ultrasensitive Timepix detector with no cooling holder allowed to collect diffraction tomography data from individual nanocrystals (size about 200 nm) and reconstruct the reciprocal space.

Unit cell determination and structure solution from the measured intensity helped to identify same crystal structure as reported by X-ray diffraction.

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Electron Diffraction Tomography analysis with TEM using ultrasensitive Timepix detector with no cooling holder allowed to collect diffraction tomography data from individual nanocrystals (size about 200 nm) and reconstruct the reciprocal space.

Unit cell determination and structure solution from the measured intensity helped to identify same crystal structure as reported by X-ray diffraction.

3D reciprocal space reconstruction of CBZ and studies crystal (about 200 nm size)

Crystal structure of CBZ solved with X-Ray diffraction

Solved crystal structure of carbamazepine using 3D electron diffraction tomography (50º continuous tilt, 3823 reflections , 0.8º resolution)

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Electron Diffraction Tomography technique by TEM microscope is particularly useful in case of polyphasic systems (several polymorphs), nm size crystals, and poorly crystalised samples

CM 30 Philips electron microscope and Timepix ultrasensitive detector(insert) at CciT Univ of Barcelona (Spain) where CBZ ED data collection was made.

Trace analysis with TEM high resolution imaging and electron diffraction

High Resolution Virtual Dark Field (VDF) in TEM is a technique that enables detection of very small trace of crystalline material; in the example shown above, trace crystals of very small sizes (eg 10nm) can be observed at very low quantity (< 0.01%).

From left to right : Virtual Dark Field (VDF) TEM high resolution image showing 10 nm resorcinol crystals (arrows) on amorphous background; corresponding ED patterns for crystallites ; individual Nicotinic acid API nanocrystal and its corresponding ED pattern

Structure characterisation (like phase confirmation) of such small crystals can be done using Electron Diffraction on individual crystallites.

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Drug polymorph structure analysis with TEM 3D electron diffraction tomography Electron Crystallography is considered as the method of choice for structure determination of nanocrystalline compounds (crystals as small as 20 nm to several microns). Such nanocrystallites reveal typically “X‐Ray amorphous” powder diffraction patterns (for sizes < 10nm) where is very difficult to identify and characterise their structures using X-Ray diffraction techniques.

Use of precession 3D electron diffraction (PED) with TEM makes possible unit cell and structure determination on individual nanocrystals. Using 3D diffraction tomography, a 3D reconstruction of the reciprocal space can be performed by tilting the sample and recording ED patterns (Fig. 1) (typically ±45° every 1°).

Collected electron diffraction (ED) patterns can be processed to precisely determine the unit cell and reveal the space group symmetry of the API crystal. Full atomic crystal structure can also be performed after collection and precise measurement of ED intensities.

From left to right: CM30 (300 KV) Transmission electron microscope (TEM), individual Carbamazepine (CBZ) API crystal, 3D reciprocal space reconstruction of CBZ crystal, CBZ Structure solved from 3D electron diffraction data (50º continuous tilt, 3823 reflections, 0.8º resolution). Cell parameters: a =7.53 Å; b =11.14 Å; c =14.06 Å; β = 92.80 º, P21/n (monoclinic). In blue structure solved by Single crystal X-Ray diffraction, in red structure solved by Electron Diffraction.

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CASE STUDY: Measurement of Powder or Lyocake Substances Made Easier - Even in Glass Vials The measurement of powder or lyocake substances is a frequently encountered and demanding task in the pharmaceutical and biotech industry.

The Chauvin Arnoux company Indatech solved it with its HypeReal system based on hyperspectral cameras from the Finnish manufacturer Specim.

Indatech´s HypeReal system enables automated, economical measurement of lyocake and powder-like substances in industries such as pharmaceutical or biotech. It drastically reduces development times of vaccines.

Image courtesy of Indatech.

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Chauchard and his team wanted to develop a solution to this problem with a system that was as easy to use as possible, based on a hyperspectral camera.

“Our goal was to make the analysis of powder-like substances as easy as operating a photocopier: it should be enough to place a sample on a microtiter plate and click a button to get the results!”

Fabien Chauchard

INNOVATIVE DESIGN The breakthrough came from imaging the sample from beneath the vial.

Looking through the base provides a flat, unobstructed view. HypeReal employs a near-infrared hyperspectral camera and spectrograph to capture detailed spectral signatures, which classification algorithms then use to identify material properties.

“We decided to measure the powderlike substances from below because this perspective offers the best overview of a flat surface and the largest possible volume,” explains Fabien.

“Checking from the side of the vials usually used in the pharmaceutical and biotech industry would be difficult because of the stickers that are usually placed there to identify the samples. The top view was also out of the question due to the plastic or metal caps used.”

At the heart of the system is the Specim FX17 camera, chosen for its compactness, robust performance across temperatures, high image quality, and GigE interface that simplifies control.

“The fact that we chose Specim as the supplier for the hyperspectral camera used made the implementation of the HypeReal project considerably easier.”

Fabien Chauchard

The InGaAs hyperspectral camera SPECIM FX17 is the basis for the HypeReal system from Indatech.

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The first tests already showed that Indatech had made the right choice, Chauchard recalls: “The tests were very encouraging right from the start and showed that the FX17 in combination with the associated lens delivered really good image quality.

We also carried out several tests to assess the robustness, e.g., by changing the temperature in the housing. However, the FX17 always provided reliable information, even under difficult conditions.”

The time savings are dramatic. For example, measuring residual moisture in a full 96-well plate using Karl Fischer titration can take weeks.

With HypeReal, the same task is completed in about two minutes, with additional quality insights delivered at the same time.

BENEFITS IN PRACTICE HypeReal offers fast, nondestructive, and contactless measurement. It can detect residual moisture in lyophilised products, verify active pharmaceutical ingredients, reveal defects such as cracks or melts, and assess homogeneity—all without toxic reagents. The platform supports different plate and vial formats, includes built-in references, and provides a user-friendly interface for data, models, and reporting.

ADOPT ION AND OUT LOOK Major vaccine manufacturers already use HypeReal, underscoring its reliability and value. Indatech sees strong potential beyond pharma and biotech, with applications in agriculture for seed analysis, in the food industry for flour quality, and even in cultural heritage fields such as document and painting examination.

By combining Specim’s advanced camera technology with Indatech’s innovative system design, hyperspectral imaging has become a powerful new tool for ensuring quality and efficiency in pharmaceutical and biotech research and production.

Want to discuss hyperspectral imaging and pharmaceutical applications?

Contact Luke Nicholls Telephone: (01372) 378822 Email: luke@qd-uki.co.uk

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Combined SEM and AFM Particle Analysis of Vitamin C Using FusionScope® Extract individual particle morphologies, surface roughness, and potential surface contaminations with correlative imaging and forcedistance curve analysis.

Modes: SEM, AFM Topography and Phase, Force-Distance-Curves Sample: Vitamin C Particles

Surface roughness of Vitamin C tablets, which can be observed by Scanning Electron Microscopy (SEM), plays a crucial role in defining its surface morphology and therefore its delivery rate and interaction time with external stimuli. Atomic Force Microscopy (AFM) then can be used to measure this roughness; however, since the size of these Vitamin C particles can range from several microns to less than hundreds of nm, the accurate positioning of an AFM probe onto specific individual particles becomes very challenging and time consuming.

There are two reasons for this. First, traditional AFM geometry only allows a top-down view of the sample surface with optical microscopy OM and so the AFM tip and direct tip interaction with the sample surface cannot be observed easily.

Second, OM in AFM cannot resolve structures below 200 nm very well (as compared to SEM).

Using FusionScope’s SEM-enabled Profile View along with its shared coordinate mapping, both challenges can be resolved simultaneously without having to change sample environment.

This reduces transfer times and contamination exposure while minimising possible timebased degradation of the particles.

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Challenges to Solve When creating tablets for health and pharmaceutical based ingestion, such as Vitamin C, information on several very important features is required. These features include Surface Morphology and Structure, Reaction to External Stimuli, Delivery Rate, and Contamination levels. To understand these features better, the particles/nanoparticles constructing the tablet are most often analysed using a range of different techniques. All in all, this nanoparticle information must provide a clear understanding of the tablet to resolve technical challenges such as the ideal delivery rate to maximise the impact of the particle (in medicinal and supplement applications). However, this process must also be done in the most efficient manner possible, for example, by using minimum resources, maximising throughput, and being very repeatable.

Current Solution Surface roughness has become a more significant characteristic of nanoparticles, including Vitamin C, due to its impact on surface morphology.

Responses and interactions to external stimuli can be understood with greater clarity, providing more precise information on exchange rate and delivery rate. The best way to measure this property, and one that has gained popularity most recently, is Atomic Force Microscopy (AFM). However, while positioning of the tip on Vitamin C particles on the scale of µm is straightforward, Vitamin C particles often can be as small as 60 nm. Therefore, positioning of an AFM using conventional optical microscopy (found on most AFMs) is very difficult to do. Additionally, since the surface of Vitamin C tablets on the nanometer scale as well as micrometer scale is very rough, positioning of the AFM using conventional approaches is very challenging resulting in contamination of the tip as well as being out of position.

Researchers in academia as well as in industry utilise a range of techniques to acquire this nanoparticle based information including Xray Diffraction (XRD), for crystallinity information, Energy Dispersive X-ray Spectroscopy (EDS) for direct chemical information, and Transmission Electron Microscopy (TEM) for porosity information.

At present, a very regularly used technique to acquire morphology and size-based information about the Vitamin C particles is Scanning Electron Microscopy (SEM). As well as direct morphology information about the particle itself, SEM can also be used to monitor morphology changes of the particle upon exposure to external stimuli — such as that of chitosan whereby Vitamin C is encapsulated by TPP-chitosan microspheres or hydroxiapetite based Vitamin C for treating bone infections. While surface structure information can be acquired through observation, size can be measured through extrapolating these images.

Our Approach In this study, we present a comprehensive investigation of Vitamin C particles utilising the FusionScope, a unique correlative analysis platform. FusionScope is a powerful tool that seamlessly combines SEM, AFM, and EDS in a single platform (and within a single user interface) offering highresolution imaging and precise measurements at the nanoscale. By combining SEM imaging, AFM topography measurements, and forcedistance curve analysis, we can extract individual particle morphologies, surface roughness, and potential contaminations on the particle surfaces. The ability to identify specific individual particles with the SEM, guide the cantilever tip to exactly the point of interest, and observe the AFM measurements using SEM-enabled Profile View, paves the way for enhanced analytical methodologies in particle research and analysis.

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Workflow Find the desired particle using the SEM

Navigate the tip using the SEM Profile View

Perform high-resolution AFM measurements

Analyse the correlative data

Figure 1: Workflow to obtain correlative AFM-SEM data on Vitamin C particles using the FusionScope. (a) Identifying target Vitamin C particle. (b) Profile View of cantilever tip positioned on particle. (c) Highresolution AFM image of particle topography. (d) Correlated AFM/SEM image of particle

In the initial phase, the targeted Vitamin C particle can be easily identified and located with the SEM (Figure 1a). Charging often plays a major role in the SEM imaging of organic particles and can impede accurate measurements. To mitigate this challenge, a beam acceleration voltage of 3.5 kV was utilised alongside working in Profile View (an 80° tilt of sample and AFM in respect to the SEM), effectively curbing the charging phenomenon experienced with these particles.

Users benefit from FusionScope’s unified coordinate system of AFM and SEM, which allows for the automated and precise navigation of the AFM tip to the desired particle.

Profile view gives direct line of sight to the cantilever tip region and ensures the positioning of the tip at the desired location on the particle (Figure 1b). High-resolution AFM measurements in amplitude modulation (AM) mode then can be performed (Figure 1c), with the SEM providing real-time visualisation of the cantilever tip movement. Both SEM and AFM data can be directly correlated in the FusionScope software, making direct data analysis easy and straightforward (see Figure 1d).

Analysing a Variety of Vitamin C Particles The SEM overview image (as depicted in Figure 2) reveals a variety of different particles exhibiting a diverse range of sizes. From notably sizable particles with diameters > 100 µm (1), to medium-sized particles with diameters > 10 µm (2), down to numerous diminutive particles (3), the spectrum of particle size is extensive.

To provide a comprehensive characterisation of these distinct particles, this study focused on analysing three particles showcasing different shapes and/or sizes to assess their surface roughness. Figure 3 displays the SEM top view of the three individual particles, whereby a different size and structure of the particles can easily be seen.

Figure 2: SEM overview image of different Vitamin C particles. Large (1), medium (2), and small (3) particles can be identified.

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Figure 3: SEM Top View of three different individual Vitamin C particles

Roughness Measurement Using the FusionScope Utilising FusionScope’s unified coordinate system for both SEM and AFM, seamless navigation of the AFM tip toward each individual particle is facilitated (Figure 4). The near-orthogonal perspective of the sample and the tip further facilitates precise positioning, particularly advantageous when approaching rough or small structures, such as Particle 3.

High-resolution AFM measurements in AM mode can then be performed on all three particles (see Figure 5). As could already be guessed from the SEM top view images Figure 3), Particle 2 has an internal structure that is not visible in Particles 1 and 3. The evolution of the AFM data reveals a sample roughness of 600 nm for Particle 1, 420 nm for Particle 2, and 150 nm for Particle 3.

Figure 4: SEM Profile View of the cantilever approaching three different particles.

This demonstrates the capability of the FusionScope to analyse the surface topography and roughness of individual Vitamin C particles easily and quickly, independently of their size or surface morphology.

Figure 5: AFM Topography data recorded in AM mode on each individual particle are shown. The blue rectangle indicates the area in which the roughness measurement was carried out. Particle roughness varies from ~600 nm (Particle 1), to ~420 nm(Particle 2), and ~150 nm (Particle 3).

Using Phase Imaging and Force-Distance Curves to Investigate Possible Contamination A common challenge encountered in particle preparation lies in the identification of specific contaminations. By combining phase imaging and force- distance curves on individual particles, FusionScope offers a solution to identify and analyze potential contaminations of the particle surface. Highresolution AFM measurements of Particle 2 unveil a distinctive cave-like structure within its topography (see Figure 6 top left).

However, it is noticeable that a phase contrast is present within the crater, indicating a different surface material of potential contamination (see Fig 6 top right).

This becomes even more visible in the overlayed image of AFM topography and phase (see Figure 6 middle). Using ForceDistance Curves on the two different areas inside the crater structure reveals that the possible non-contaminated part (Figure 6 bottom left; Position 1) exhibits a greater hardness compared to the contaminated part (Figure 6 bottom right; Position 2). In addition, Position 2 also displays a stronger adhesion compared to Position 1. These two characteristics of the force-distance curves indicate that Particle 2 consists of different materials.

Figure 6: (Top) High resolution AFM topography and phase image of Particle 2 indicates a possible contamination.

(Middle) Overlay of AFM topography and phase image revealing the two different materials in the crater structure. (Bottom) Force-Distance Curves at Position 1 and Position 2 indicating different hardness and adhesion.

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Preventing Misinterpretation As the cantilever approaches the sample of Force-Distance Curves surface (1) it contacts the sample (2). Then the The interpretation of force-distance curves to gain insights into the mechanical properties of a sample presents considerable challenges.

Especially for small non- flat structures (e.g., Particles, Fibres, Nanowires) the cantilever can move or damage the samples during the process of obtaining the force-distance data.

Hence, the ability to observe the cantilever's movement during the force-distance curve becomes indispensable for identifying potential artifacts. Such an artifact is illustrated in Figure 7, where the whole forcedistance curve is observed in Profile View with the SEM.

Figure 7: SEMimages1-4 show the position of the cantilever during the acquisition of a force-distance curve.

To illustrate the movement of the particle during the forcedistance curve, horizontal dashed lines were added to the SEM images. The forcedistance curve was divided into three sections (a-c). The force-distance curve (shown in (a)) in the range between images 1 and 2 shows constant deflection of the cantilever during the initial approach to the particle surface.

cantilever first pushes the particle down onto the surface (3) before executing the real mechanical probing of the particle (4). This phenomenon culminates in the cantilever bending, as evidenced by the red curve (Figure 7b), a distortion that could inadvertently lead to misinterpretation of the slope as a real mechanical property of the sample.

By combining force-distance measurements with SEM Profile View, FusionScope can circumvent these misleading results and provide a comprehensive analysis of mechanical properties of small objects.

The range between 2 and 3, on the other hand, shows an increase in the deflection (b).

This deflection is because the cantilever pushes the particle downward onto the surface and does not exhibit any real mechanical data. The green curve in (c) shows the sequence of the forcedistance curve between the SEM images 3 and 4. A very steep increase can be noticed since the particle is now completely pushed onto the sample surface. The complete force-distance curve is shown in figure (d).

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FusionScope™ from QD Microscopy Easy to use Correlative AFM with SEM Microscopy Platform Material Characterisation Quality Control learn more Failure Analysis Nanostructures

OUTCOMES We have reduced transfer time between different measurement systems by having all measurements conducted within one sample chamber and user interface.

We have combined different measurement modes into a single system, making it simpler to use and learn, therefore you do not need an expert in each individual technique. A single user can learn fairly quickly how to use all the techniques on one platform.

We have made the acquisition process of data, with AFM, very easy by being able to see the cantilever tip approaching the sample surface– minimising measurement setup time as well as contamination or tip damage inspection time.

We are able to perform measurements correlatively, using a joint coordinate system, thereby getting direct like-for-like data independent of time, at any angle of interest and very localised scales.

We have created an easy to maintain system with strong service and support.

Want to discuss FusionScope and the opportunities it offers your research?

Contact Dr. Luke Nicholls Telephone: (01372) 378822 Email: luke@qd-uki.co.uk

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CHARACTERISE SUBWAVELENGTH PARTICLES USING AFM AND ENERGY DISPERSIVE SPECTROSCOPY (EDS) ALONG WITH PROFILE VIEW Modes: SEM, AFM Topography and Phase, EDS Sample: Superparamagnetic Particles

Easy to use Correlative AFM with SEM Microscopy Platform Fig 1. Topography and phase images of a particle (600 x 600 nm with 600 x 600 pixels in the x- and y-axis). The surface roughness (RMS) shown by the dashed square (250 x 250 nm) on the image above is 25.01 nm

For this example, we use carboxyl (COOH) group functionalised magnetic particles as a model system to demonstrate FusionScope’s imaging capability. The FusionScope allows us to image the particle’s surface with picometer resolution.

The wide field of view of the SEM is employed to localise particles for AFM imaging.

Combine the complementary strengths of AFM and SEM like never before! The FusionScope fully integrates a wide range of advanced AFM measurement techniques with the benefits of SEM imaging.

Seamlessly image your sample, identify areas of interest, measure your sample, and combine your imaging data in real time.

The topography and phase data from the AFM permit high-resolution inspection of the particle surfaces. We then use the correlated coordinate system and correlative in-situ measurement modes to capture relational AFM and SEM data.

Higher-resolution AFM images from a single particle are shown in Figure 1. Finding particular particles of this size to take AFM data would be extremely difficult without FusionScope's correlated coordinate system and Profile View.

Read the Article

Discover the FusionScope

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HOW PIFM OUTPERFORMS CONVENTIONAL TECHNIQUES SURFACE-SENSITIVE CHARACTERISATION OF ATOMIC LAYER DEPOSITION FILMS ON DRUG PARTICLES

INTRODUCTION Coating the surface of acetaminophen particles with Al₂O₃ and ZnO via atomic layer deposition (ALD) has been shown to effectively modulate drug release rates [1].

The success of these approaches fundamentally depends on the uniform and conformal application of ALD coatings [2].

This, in turn, raises a critical question: how can we rigorously confirm the successful formation of an ultrathin layer? Addressing this question necessitates a technique capable of providing surface-sensitive chemical information with high spatial resolution.

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CURRENT METHODS While surface-sensitive techniques such as TOF-SIMS can detect the presence of Al₂O₃, they are limited to identifying its existence rather than providing comprehensive information on its distribution or coverage across the particle surface.

Merely establishing the presence of Al₂O₃ at isolated locations is insufficient for validating uniform ALD deposition, thereby excluding TOF-SIMS and similar methods from consideration for this application.

Techniques such as SEM/EDS and TEM-EDS may also be considered; however, the integration of their signals over significant sample thicknesses results in a lack of true surface sensitivity, especially for characterising ultrathin or single-molecule layers.

In practice, SEM/EDS and TEM-EDS are far more likely to yield dominant drug substrate and elemental signals, with little to no sensitivity to the ALD overlayer.

Figure 1: Top left and right are EDX images of the sample after ALD. Bottom spectra are ATR spectra from untreated acetaminophen sample and 50 cycles ALD Al2O3 on an acetaminophen sample [1].

ATR-FTIR is another candidate technique, utilising an evanescent wave generated at the interface of a high-refractive-index crystal to probe the sample. Figure 1 demonstrates the application of ATR-FTIR to Al₂O₃ ALD on acetaminophen. However, as evident from the spectra, ATR-FTIR lacks the required surface sensitivity for this application. The spectra of untreated acetaminophen and that of acetaminophen after 50 ALD cycles are essentially indistinguishable.

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UNIQUE PIFM APPROACH Clearly, the techniques discussed in the previous section do not have the spatial resolution or surface sensitivity to investigate ALD films. In contrast, photo-induced force microscopy (PiFM) offers a uniquely suitable solution.

PiFM’s capability to fix the excitation laser at a specific wavenumber and raster scan the sample enables the collection of chemical maps with exceptional sub 5 nm spatial resolution.

Once regions of interest are identified, single-point, surface-sensitive wavenumber sweeps can provide detailed chemical spectra from the very surface.

Figure 2 presents PiFM data for Al₂O₃ films on an unnamed drug molecule.

Samples A and F possess unknown levels of Al₂O₃ ALD film on the drug particles. AFM topography images for both samples reveal surface roughness or aggregation features approximately 10 nm in height.

Because the surfaces look so similar, the topograhy cannot provide any hints to determine if an ALD monolayer has been formed. Therefore, we can apply PiFM to gain chemical data on the sample. By tuning the PiF excitation laser to 850 cm−1 (the Al₂O₃ signature), PiFM imaging reveals a strong Al₂O₃ signal in sample F and a weak signal in sample A. Conversely, imaging at 1235 cm−1 (the drug IR signature) yields a strong drug signal in sample A and a weak signal in sample F. These results indicate that sample F is coated with a uniform ALD monolayer, whereas sample A exhibits none or very little ALD coverage. Therefore, PiFM allows us to gain localised chemical information about these surfaces, which is not something that any of the other analytical techniques considered could provide for this application.

Figure 2: AFM topography images, 850 cm−1 PiFM chemical absorption maps, and 1235 cm−1 PiFM chemical absorption maps for samples A and F

In addition to the PiFM chemical maps, we can corroborate these findings using PiF-IR point spectra and comparing them to various references as shown in Figure 3. Each PiF-IR spectrum in Figure 3 is an average of 6 spectra acquired with a ~150 nm pitch. The FTIR control spectrum for Al₂O₃ (grey) displays no distinct peaks and a gradually increasing signal starting at 1100 cm−1.

The control uncoated drug particle spectra, acquired via ATR FTIR (pink), shows the IR peaks associated with the drug molecule. The averaged PiFIR spectrum taken on the uncoated drug particle (blue) shows peaks that agree well with the peaks from the pink reference spectrum on the uncoated drug material ATR FTIR spectrum. The PiF-IR spectrum taken on Sample A (green spectrum) exhibits features matching the uncoated drug particles, with no increase in peak strength below 1100 cm−1.

Figure 3: Averaged PiF-IR spectra from an Al2O3 substrate (grey), an ATF FTIR spectrum from the uncoated drug (pink), Averaged PiF-IR spectra from an uncoated drug particle(blue), and Averaged PiF-IR spectra from Sample A (green) and Sample F (red).

This is consistent with the low signal intensity in the PiFM image at 850 cm−1 in Figure 2. Sample F’s PiF-IR spectrum (red) displays a pronounced Al₂O₃ signal, as indicated by the broad, rising baseline from 1100 cm−1.

This is indicative of a more continuous Al2O3 layer, as supported by Figure 2.

Notably, faint drug-related peaks are still observable in sample F, demonstrating PiFM’s ability to probe through ultrathin ALD films.

Ultimately, these point spectra are consistent with Figure 2, strengthening the spotty and complete monolayer diagnoses we made using the single wavenumber PiFM images.

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Therefore, combining PiFM imaging with PiF-IR point spectroscopy allows detailed and accurate chemical characterisation at points of interest determined from topographical images.

In summary, PiFM stands out as the only technique capable of simultaneously delivering surfacesensitive, spatially resolved, and chemically specific data on ALD coatings.

The integration of these three critical dimensions of information is essential for rigorously validating the uniformity and integrity of ALD surfaces.

While the data presented in this application note used an unknown drug molecule as the substrate, a similar analysis can be performed for any combination of substrate and thin film (for example, inorganic or organic substrate covered with inorganic or organic film in any combination).

Want to discuss PiFM imaging and pharmaceutical applications?

Contact Dr. Shayz Ikram Telephone: (01372) 378822 Email: shayz@qd-uki.co.uk

Molecular Vista “Vista 200”

Molecular Vista “Vista 300”

For nano‑IR chemical analysis

Industry-leading automation

With a 200 mm × 200 mm sample stage Vista 200 is perfect for analysing wafers or photo masks. Can be configured with either a vacuum chuck with magnets or a specialised EUV mask holder.

This instrument allows access to features on samples as large as 300 mm with PiFM & PiF-IR. AutoPiFM and AutoAlign make Vista 300 the most advanced nano IR instrument ever made.

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PiF microscope

PiF microscope

Precision in Powder Analysis: Electron Diffraction for Milled Pharmaceuticals

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SOLVE PROBLEMS

BY SAVING TIME, RESOURCES AND MATERIAL

MILLING Milling is widely utilised in the pharmaceutical industry, serving several critical applications, including enhancing solubility and bioavailability, particle size reduction, controlled release formulations. The product of milling can sometimes be difficult to analyse for traditional techniques such as PXRD or X-ray. We show here how electron diffracion is ideal for analysis of milled products.

Milling enhances the solubility and bioavailability of poorly soluble drugs by producing amorphous materials that dissolves more readily than their crystalline counterparts.

Milling also plays a key role in particle size reduction, ensuring uniformity in active pharmaceutical ingredients (APIs) and excipients, which is vital for consistent drug formulation and enhanced drug absorption. Additionally, milling is crucial for developing controlled-release formulations, as it helps achieve the desired particle sizes and distributions necessary to maintain therapeutic drug levels over extended periods.

The ELDICO ED-1 device is dedicated solely to electron diffraction ). It is a user-friendly device that investigates crystalline samples with particle sizes below 1000 nm

Overall, milling not only improves the physical and chemical properties of pharmaceutical compounds but also supports the development of advanced drug delivery systems, ultimately contributing to more effective and reliable medications.

Analysing the powder using X-ray powder diffraction (XRPD) is vital for milled samples for several reasons: 1. Confirming Amorphous State 2. Detecting Residual Crystallinity 3. Ensuring Batch Consistency 4. Regulatory Compliance However, XRPD has a detection limit of 1-2%, which can sometimes overlook low levels of residual crystallinity or impurity phases.

To address this limitation, electron diffraction has emerged as a promising solution. With its higher sensitivity, electron diffraction can detect even minimal crystalline residues, ensuring a more accurate characterisation of the milled materials. This advancement supports the development of more effective and reliable pharmaceutical formulations.

The ELDICO ED-1 from ELDICO Scientific is optimised for this purpose as it has the ability to analyse powders fully automated using the so called electron diffraction crystal mapping method.

Carvedilol, comparison ball milling and melt quenching In collaboration with the Research Center Pharmaceutical Engineering GmbH (RCPE) in Graz, Eldico Scientific conducted a case study to compare different sample preparation methods.

Carvedilol was used as an example to assess two different preparation techniques for amorphous APIs, melt quenching and ball milling. To investigate the resulting crystallinity from both preparation techniques, samples were prepared on a TEM grid. For each sample about 100 particles were tested using the automated crystal mapping method.

Fig. 1: Results electron diffraction mapping

CASE STUDY Whereas the sample from melt quenching was fully amorphous, crystallinity was still found to be present in samples prepared by ball milling (Fig. 1). Continuous rotation data collection allowed the determination and matching of the unit cell with Carvedilol.

The results indicate that ball milling time would need to be increased to ensure 100% amorphisation.

Conclusion Electron diffraction crystal mapping emerges as a powerful tool for precise characterisation of milled pharmaceutical samples. This technique excels in detecting crystalline phases even at extremely low levels of detection, making it invaluable for assessing the amorphous nature of milled materials. By combining particle screening with continuous rotation diffraction, electron diffraction crystal mapping identifies and identifies crystalline impurities and unknown crystal forms within pharmaceutical formulations. This capability is crucial for ensuring product quality and efficacy, as well as meeting stringent regulatory standards in the pharmaceutical industry.

MOLECULAR VISTA

Offering the highest resolution nanoscale chemical analysis instruments available

FEATURES Sub-5 nm IR spatial resolution Unique chemical identification capability

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Built for industry Maximise efficiency, minimise disruption The 300 mm wide sample access door minimises thermal disturbances that cause drift while allowing you to maximise throughput. With Molecular Vista’s forward moving stage and user friendly head mount, both the tip and sample can be easily swapped without opening the entire enclosure.

Self-contained system No need for a special environment. Vista 200 is complete with built in vibration isolation, a temperature controlled acoustic enclosure with 0.1 ºC precision, and a fully enclosed beam path with dry air.

Streamlined processes Vista’s included data analysis software allows your team to go from measurements to presentation-ready results quickly. Tight integration with this data acquisition software allows you to get the answers you need in real time. The scripting API allows custom recipes, and files are open so integration with 3rd party software is easy.

Exceptional AFM performance With a 120 µm xy-scanner, 12 µm sample z-scanner, and integrated vibration isolation, the AFM is top notch. The highbandwidth dual-z feedback system allows true non-contact AFM for IR PiFM.

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MicroCT of Pharmaceuticals and Pharmaceutical Packaging MICROCT AND MICROXRF HAVE BECOME INTEGRAL TOOLS FOR INSPECTING PHARMACEUTICAL PACKAGING AND DEVELOPING NEW DRUG DELIVERY METHODS

Pore network of the polymer sustained release coating for a pill. Imaged by Sigray’s patented multi-spectral x-ray source in EclipseXRM.

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Sigray’s EclipseXRM enables rapid non-destructive 3D analysis of pharmaceutical formulations and their packaging. The AttoMap XRF microscope provides mapping of inorganic materials within a tablet, aiding in the understanding of active ingredient distribution and the detection of contaminants.

3D x-ray microscopy (also known as microCT) is increasingly recognised as an essential tool for both failure analysis and the R&D of new pharmaceuticals. For example, ultrahigh-resolution imaging (<300nm spatial) of pore networks and active ingredients provides insight into drug release mechanisms.

Feedback on internal structure can accelerate time-to-market during the research and development phase.

Even after a product has been successfully developed, microCT remains valuable for identifying failures in pharmaceutical packaging.

EclipseXRM provides the highest-resolution and highestcontrast images for the wide range of samples encountered in a pharmaceutical central laboratory—from polymeric drugs to packaged lyophilised (“lyo”) cakes.

3D view and cross-sections of a Pepcid AC within its packaging

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CASE STUDY: Next Level of Pharmaceutical Inspections

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Q

uality assurance is of utmost importance in pharmaceutical products. Striving for the world where drugs are reliably monitored and tracked, SEA Vision has introduced hyperspectral inspection solution HARLENIR, the combination of SEA Vision software and Specim camera technology which reveals the chemical composition of a pharmaceutical product and guarantees 100% inspection coverage, accurately and reliably. SEA Vision chose Specim FX17 since it was the only hyperspectral camera on the market to meet their requirements of speed and spectral resolution.

SEA Vision is a leader in the pharma industry and a pioneer in applying artificial vision in pharmaceutics quality inspection.

It was founded in Italy, Pavia as a university spin-off in 1995, and produces top-quality vision systems solutions to Pharmaceutical companies and packaging machine manufacturers.

Throughout their existence, SEA Vision has been striving to provide the highest quality vision systems for the pharmaceutical industry. They rely on the latest innovations and the best technologies to improve their product portfolio for quality inspection and to comply with regulatory guidelines. This goal is supported by their technical and R&D department’s high competence in software algorithms which meet the unique needs of Pharmaceutical customers and OEMs business.

All this has made SEA Vision the leader of their field.

QUALITY BEYOND THE VISIBLE INSPECTION To provide the pharmaceutic industry with the highest quality best solutions, SEA Vision has introduced a new generation of blister inspection technology that goes beyond visible inspection: hyperspectral imaging.

Aerial shot of Pavia, Italy

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Hyperspectral imaging provides a reliable way to perform a chemical composition inspection of pharmaceutical products during the production process. This is particularly important with products that are visually similar but have different proportions of active pharmaceutical ingredient (API).

With hyperspectral imaging, each tablet or capsule can be quickly and reliably identified from their active ingredients.

100% of the products can be inspected, online, in real time.

The chemical analysis allows: qualitatively to identify pharmaceutical products with different active principle. quantitatively to identify pharmaceutical products with a different dosage, and to measure the uniformity of the active ingredient distribution

“In an era of increasing regulation and quality control, the industry needs to improve inspection methods. Systems like HARLENIR provide extra peace of mind for drug manufacturers” Vittorio Calbucci from SEA Vision R&D Department

“To this aim,” continues Vittorio Calbucci, “we installed HARLENIR system on a blister machine provided by one of the most important Italian OEM. The idea was to check the presence of foreign tablets with different API respect the tablets with the right chemical compounds, and tablets with the same API but different dosages.”

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“ We always choose the best

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technologies for our products, and Specim camera fits our needs.”

Vittorio Calbucci

WHY SPECIM? Inside SEA Visions HARLENIR solution is Specim FX17 camera.

“We chose the system which best satisfied the customers’ requirements,” says Calbucci.

Before deciding on Specim FX17, SEA Vision evaluated several different technologies versus hyperspectral imaging, such as THz spectroscopy or the combination of a standard vision system with a fibre optic spectrometer.

However, with the first technology, the resolution and speed were too low while with the second solution it was not possible to inspect the whole sample.

Hyperspectral imaging was chosen as it is possible to perform chemical analysis in a fast way and with high spectral resolution in combination with colour camera imaging which provides an accurate check of pictorial and geometrical parameters of the objects.

Since the acquisition rate is so high, the hyperspectral camera can be installed directly over the production line. It will image the targets as they pass the camera on the conveyor and collects data of the whole surface of the target, covering every pixel of every inch.

The production line where HARLENIR with Specim FX camera is used produces up to 3000 tablets per minute, which requires an acquisition frequency of close to 500Hz. Specim FX17 frame rate of 640 Hz is good enough to cover the whole production. Specim FX17 compact size also allowed it to be installed in a small space without compromising the technical quality.

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The Specim FX camera is ready-to-use, and SEA Vision was able to perform the first chemical analysis already on the first days after installation. For the few technical glitches, there was good and timely support available both directly from Specim as well as from their Italian distributor.

SPECIM FX17 cameras are designed for industrial use, and every camera is calibrated to provide the same output.

Thus if there is a need for maintenance, the camera can be swapped to a replacement unit on the fly without compromising the production.

Hyperspectral cameras let you see beyond the visual inspection - they reveal the invisible

WHAT IS TO COME? In addition to the tablet inspections, SEA Vision has already tested the Specim FX17 camera and hyperspectral imaging on other applications, where it worked as expected. In the future, they expect hyperspectral imaging to help to widen their product portfolio and develop new solutions.

To discuss your application or area of research, please get in touch with our Sales Manager, Dr. Luke Nicholls.

Call (01372) 378822 Email luke@qd-uki.co.uk

Non-destructive, rapid, and reliable inspection with close to 100% accuracy Inspect 100% of the product stream in real-time Measure the presence, amount, and distribution of API Avoid mix-ups and improve product quality and safety High frame rate High spectral resolution Small size Ready-to-use Good and timely support

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