Open Access Article
Erwin
Winkler Martinez
a,
Ali
Tfayli
a,
Thomas
Nappez
b,
Jean-Philippe
Michel
c,
Douglas N.
Rutledge
a,
Pierre
Chaminade
a and
Sana
Tfaili
*a
aUniversité Paris-Saclay, Stratégies d'investigations analytiques: Médicaments, bioMatériaux, tissus et Matrices biologiques, 91400, Orsay, France
bConnected Physics, Fabrication d’équipements médicaux, 92220, Bagneux, France
cUniversité Paris-Saclay, CNRS, Institut Galien Paris-Saclay, 91400, Orsay, France
First published on 26th March 2026
The prevalence of substandard and falsified (SF) drugs requires the development of effective identification and differentiation methods to distinguish genuine from SF medications. SF drugs may have harmful consequences, including increased morbidity and mortality. Near-infrared (NIR) spectroscopy is a well-established technique that has demonstrated its effectiveness in detecting poor-quality medicines. It provides a rapid, specific, and non-destructive method for identifying SF. NIR spectroscopy may be applied in hyperspectral imaging, multispectral imaging, or point spectrum recording. Spectral NIR data, advanced data treatment, and chemometrics have proven to be essential in drug analysis. This project aimed to develop a low-cost visible and near-infrared (Vis-NIR) handheld and portable multispectral imager for fast drug analysis and detection of SF drugs. Low-cost spectrophotometers of this sort in low-income countries offer a cost-effective and quicker alternative to traditional laboratory methods (e.g., mass spectrometry). For this, we analyzed paracetamol tablets (test samples) and commercial tablets. The developed device employs multispectral imaging (MSI). We processed data using Principal Components Analysis (PCA) and Independent Components Analysis (ICA). The preliminary results suggest that our prototype has promising potential, with the ability to differentiate effectively between tablets with different compositions.
The MSI approach allows real-time and non-destructive analysis. It provides spatial and spectral information on sample composition, making it suitable for laboratory and on-site applications.5,11 Studies show that MSI is a cost-effective and faster alternative to traditional hyperspectral imaging. Hyperspectral imaging can be used in the pharmaceutical industry to determine the homogeneity of samples.12 Compared to MSI, it provides more information since it covers the entire spectrum, though at the cost of longer data processing times. MSI covers a few wavelengths and requires less time and computing power without compromising the relevance and quality of the data.2 MSI research in the pharmaceutical field remains limited to studies addressing drug quality assessment.1,3,6 The few published studies exploring multispectral imaging were in the ultraviolet (UV) range rather than NIR wavelengths.5,8
As it reveals physico-chemical properties, NIR imaging for drug analysis11,13 is a promising method for ensuring therapeutic efficacy and patient safety. Vibrational spectroscopy techniques, such as NIR and Raman, are well known for the analysis of medicinal herbs and pharmaceuticals.14–21 However, few studies have illustrated imaging applications.11
NIR spectroscopy, spanning wavelengths from 780 to 2500 nm, captures molecular vibrations that offer insights into drug samples’ chemical and physical properties.15,22 By comparing NIR spectra from genuine drugs with those of suspicious samples, chemometric analysis can predict concentration discrepancies and variations in excipients, aiding in the detection of substandard and falsified (SF) drugs.23–26 Infrared spectroscopy – when combined with multispectral imaging – enables more detailed analysis of APIs and excipients, providing robust solutions for drug quality assessment and falsified drug detection.
Substandard and falsified drugs frequently appear in the global market, especially in low- and middle-income countries. The World Health Organization estimates that SF drugs constitute more than 10% of the world market, underscoring the need for practical analytical solutions.23,27 Falsified medicines are intentionally misrepresented in composition, identity, or origin, while substandard medicines are legitimate products that fail to meet quality standards due to production errors or negligence.27,28 Both pose serious risks to public health, contribute to drug resistance, and increase morbidity and mortality.16 Bakker et al. showed all the possible analytical methods that can be applied to identify SF drugs, showing their advantages and limitations.29 Accessible, field-deployable technology is vital, particularly for resource-limited areas. Lab-based methods (heavy benchtop devices), such as high-performance liquid chromatography (HPLC) and mass spectrometry, provide high sensitivity and selectivity, but are often not affordable and remain complex to implement in some countries.2,26,29–31
The literature describes the contribution of NIR and Raman vibrational methods in the fight against SF drugs.26 The main advantage of these methods is their speed. In their study in 2005, Rodionova et al., pioneering users of NIR for SF drug analysis, called it an “express method”.32 The instrumental development of handheld and portable devices accompanied the use of vibrational spectroscopy for this purpose.21,23,27,31,33–35 Field studies generally use handheld or portable instruments to acquire point spectra. To our knowledge, no field-study addresses the use of imagers for SF drug identification.
Portable or handheld devices are designed to be simple to operate. NIR portable or handheld devices allow on-site drug analysis and provide quick spectral information that can be easily interpreted to verify authenticity. On-site analysis allows the possibility of bringing the devices to the sample. On-site analysis helps reduce time and make fast decisions for production lines and on-site investigations.29,36 Zambrzycki et al. evaluated six handheld or portable devices for point spectra (NIR or Raman), discussing the limitations and advantages of each.37 They presented an interesting perspective on the cost of devices, as well as all the extra charges that also add up (such as consumables, computers, etc.). None of these devices is currently in use for on-field identification of SF drugs due to their high cost or low performance. In the literature, all current devices being evaluated are for point spectra, with only one imager device, the CDx,38 being identified, which remained at the proof-of-concept stage and was never commercialized.39
In the present work, we aim to develop a low-cost, portable visible and near-infrared (Vis-NIR) multispectral imager for fast drug analysis and identification of SF drugs. The use of low-cost portable spectrophotometers in low- and middle-income countries offers a cost-effective and quicker method than alternative standard laboratory methods. The first tests were conducted on two commercial samples – Doliprane® and Pentasa® –, coated tablets, and paracetamol tablets. Paracetamol samples were produced in the laboratory to ensure precise percentages of API and excipients, following European Pharmacopeia guidelines for pharmaceutical characterization. Two different formulations, 0% API and 100% API, respectively, were prepared to emulate SF products. Six samples were taken for each formulation – A16 to A21 for 0% and E23 to E27 for 100% – to ensure repeatability. The device was also tested to analyze commercial tablets, including Doliprane D01, Pentasa P01, and coated tablets R01. We defined a range of spectral bands using 13 LEDs spanning from 450 nm to 1085 nm to build the first version of our prototype. For data treatment, we used chemometric tools and multivariate analysis. The potential of our portable MSI prototype, as an effective solution for low-cost and rapid drug analysis, was demonstrated.
| Category | API | Excipient | Excipient | Lubricant | Disintegrant |
|---|---|---|---|---|---|
| Supplier | RHODAPAP® | JRS PHARMA® | Cooper® | Cooper® | SPCI® |
| Chemicals | Paracetamol (grams) | Emcompress (grams) | Wheat starch (grams) | Magnesium stearate (%) | Explotab (%) |
| Test tablet 100% API (E) | 100 | 100 | 50 | 1 | 2 |
| Samples E22–E27 | |||||
| Test tablet 0% API (A) | 0 | 100 | 50 | 1 | 2 |
| Samples A16–A21 |
Samples were placed on a plate below, and images were recorded using each LED alone once the camera parameters were set. The exposure time was set via shutter control (acquisition time), and the sensor signal from each pixel is processed to correct the offset and multiply the data by the corresponding gain (intensity of brightness). Each LED has its own combination of acquisition time and gain (see the SI section and Table S1); acquiring each image took between 10 and 15 seconds (an image per LED is acquired). All experiments were conducted inside a lightproof enclosure. Multiple tablets of varying physicochemical properties were also tested (Section S1, SI data). We also recorded images with all LEDs in the visible range turned on, with LEDs in the NIR range turned on, and with all the LEDs on for visual inspection.
A few spectra originating from the 0% API tablet formed a separate cluster in the PCA plot (the region between 0 and 200 in PC1). This cluster corresponded to spectra located at the tablet borders. By isolating these outlier pixels, we generated an image and a corresponding mean spectrum. This analysis confirmed that the outlier pixels were associated with the tablet edges (Fig. 2). These pixels were subsequently identified and removed from all images before starting the processing. PCA was used for both pre-processing and processing.
Statistical analyses including histograms for data distribution visualization, t-tests for group comparisons (each group included 0% and 100% API test tablets), and Cohen's d to measure the distance in each group were performed.
We also ran PCA and ICA on reconstructed images for LEDs in the visible range alone, and then on reconstructed images for LEDs in the NIR range alone. Similarly, we extracted the loadings of the principal component showing the separation in PCA results, and of the independent component showing the separation in ICA results. This time, the loadings of these PC and IC covered the LEDs in visible or in NIR ranges. We applied also statistical analyses including histograms for data distribution visualization, t-tests for group comparisons (each group included 0% and 100% API test tablets), and Cohen's d to measure the distance in each group.
Data processing was done on six tablets with 0% API (labelled A and numbered A16–A21) and six tablets with 100% (labelled with the letter E and numbered E22–E27), using only the images with the LEDs from 450 to 970 nm.
Since the image captured by the device is larger than the tablet and thus contains useless information, we cropped the images to the tablet's dimensions. Cropping the images allows us to focus on the region where the sample is located and extract relevant intensity data. All the images were cropped to the same dimensions. This pre-processing was followed by PCA to remove outliers. As shown in Fig. 3, each image provides spectral and spatial information, capturing variations in intensity across wavelengths and space. The multispectral image of each tablet was generated by overlaying the images recorded from each LED alone. This created a data cube. Images from each data cube were concatenated before PCA and ICA processing. PCA and ICA were conducted on data cubes in pairs.
The optimal number of principal components (PCs) was found to be 5, using the random PCA method.31Fig. 4 presents the scatter plot for the first and second component (samples A19 and E25), showing a clear separation between 100% (Yellow) and 0% (Blue) tablets. PC2 separates almost completely the two groups while PC1 also gives a partial separation; this is corroborated with the 1D score plot of component 2 (Fig. 4). As illustrated in the plot images in the supplementary data (Fig. S12) and the score plots (Fig. S14). This indicates that these two PCs contain information that helps us distinguish between both samples. Fig. 5 presents the PC2 image plots, visualizing the intensity difference between the two samples. The bar plot in Fig. 5 shows the contribution of each LED in the selected range. The highest contributions are obtained with positive values from 450 nm (blue light), 610 nm (red light) and 810 nm (NIR) light, indicating a higher signal from the 0% tablets for these three wavelengths, followed by negative values for 851 nm, 940 nm and 970 nm (NIR) light, indicating a higher contribution from the 100% API tablet (SI Fig. S12 shows PC1–PC5). Fig. 6 and 7 show the comparison between a Doliprane® tablet and a 100% API tablet. The scatter plot (Fig. 6) presents the first and second components, showing a clear separation between 100% API test (green) and Doliprane® (red) tablets. For Doliprane® compared to the 100% API test sample, the results showed the highest positive contributions at values 450 nm (blue light), 810 nm (NIR) and 870 nm (NIR), indicating a higher signal from the Doliprane® tablet for these three wavelengths, followed by negative values for 851 nm, 940 nm and 970 nm (NIR), indicating a higher contribution from the 100% API tablet (Fig. 7).
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| Fig. 6 Scatter plot illustrated for PC1 and PC2 for Doliprane® (D01) and 100% API test tablets (E25). Each color represents a tablet: red (Doliprane®) and green (100% API test tablet). | ||
Fig. 10 and 11 show the comparison between a Doliprane® tablet and a 100% API test tablet. The scatter plot (Fig. 10) presents the sixth and seventh components, showing a clear separation between 100% API test tablets (green) and Doliprane® (red) tablets. The highest positive contributions at values 450 nm (blue light), 810 nm (NIR), 870 nm (NIR) and 890 nm (NIR) indicate a higher signal from the Doliprane® tablet for these four wavelengths (Fig. 7).
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| Fig. 10 Scatter plot illustrated for IC4 and IC3 for Doliprane® (D01) and 100% API test tablet (E25). Each color represents a tablet: red (Doliprane®) and green (100% API test tablet). | ||
The presented home-built prototype – which uses LEDs and a monochromatic camera to capture multispectral images – demonstrated its potential (to differentiate samples) through a multivariate analysis. Both methods, PCA and ICA, helped to differentiate between 0% API (A16–A21) and 100% API (E22–E27) formulations. Moreover, the comparison between Doliprane® (D01), Pentasa® (P01), in SI Fig. S4, S5, S8 and S9, and red-coated tablet (R01), in SI Fig. S6, S7, S10 and S11, with our 100% API test tablet (E25) formulation showed promising results. While both PCA and ICA have their mathematical foundation, both provided relevant information. First, to discriminate between the samples and second, to understand which LEDs provided the most relevant information. Given the similar outcomes, using a single chemometric approach may be sufficient in future applications, especially when it comes to fast drug analysis. It is worth mentioning that when using ICA, a higher number of LEDs and stronger intensity signals were involved in discrimination. Comparing the signals from both methods could guide future optimization of LED selection and system configuration.
Overall, integrating a multivariate analysis, employing only a few wavelengths, a low-cost, image-based MSI camera offers a practical fast solution for drug control. Notably, while many commercial systems are limited to point spectra or designed for different types of samples, our setup enables direct image capture (MSI). With minimal spectral input and straightforward chemometric techniques, we could differentiate between tablets with close but different formulations.
Fig. 12A and B illustrate the histograms and Cohen's d-test values using the full dataset covering all LEDs. The pixel intensities for PC2 or for IC3 loadings are shown in these figures. The analysis of the outputs – for the second principal component (PC2) and third independent component (IC3) – highlighted the separation between samples, with d-values equal to 1.9466 (pixel intensities for PC2 loading) and 1.9565 (pixel intensities for IC3 loading).
Fig. 12C and D illustrate the histograms and Cohen's d-test values using the visible range (450, 525, 610, and 750 nm LEDs). When running PCA and ICA on data in the visible range, PC2 and IC1 loadings appeared to be the components separating between the groups. As illustrated, pixel intensities for PC2 or for IC1 loadings are separated between samples, with d-values equal to 1.7191 (pixel intensities for PC2 loading) and 1.7392 (pixel intensities for IC1 loading).
Fig. 12E and F illustrate the histograms and Cohen's d-test values using the NIR range (810, 851, 870, 890, 930, 940 and 970 nm LEDs). When running PCA and ICA on data in the NIR range, PC2 and IC1 loadings were found to be the components separating between the groups. As illustrated, pixel intensities for PC2 or for IC1 loadings are separated between samples, with d-values equal to 1.8002 (pixel intensities for PC2 loading) and 1.8089 (pixel intensities for IC1 loading).
The distance measurement with Cohen's d-test revealed significant separation between the two groups in all pixel's intensities for PCA and ICA, having the highest value in the whole data set and the lowest value of separation in the visible range.
The t-test (p-value) showed a significant difference for the whole data set (pixel intensities for PC2 and IC3 loadings), visible range (pixel intensities for PC2 and IC1 loadings), and NIR range (pixel intensities for PC2 and IC1 loadings).
The results support the idea that recording multispectral data provides more discriminative information than working only in the visible range. It is important to note that our device's camera is not a standard RGB camera. Our camera covers a wider range of the spectrum, allowing us to extract spectral information beyond the visible range. We need to continue working on our portable prototype: optimizing, improving and acquiring images with multiple samples.
These initial findings highlight the novelty of the device and its potential for further development in identifying SF drugs. The optimization and development of this low-cost device could provide valuable support in detecting substandard and falsified drugs in low- and middle-income countries.
Additional data are available from the corresponding author upon request.
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