Performance Characteristics of Mass Spectrometry-Based Analytical Procedures for Quantitation of Nitrosamines in Pharmaceuticals: Insights from an Inter-laboratory Study.

With the discovery of carcinogenic nitrosamine impurities in pharmaceuticals in 2018 and subsequent regulatory requirements for risk assessment for nitrosamine formation during pharmaceutical manufacturing processes, storage or from contaminated supply chains, effective testing of nitrosamines has become essential to ensure the quality of drug substances and products. Mass spectrometry has been widely applied to detect and quantify trace amounts of nitrosamines in pharmaceuticals. As part of an effort by regulatory authorities to assess the measurement variation in the determination of nitrosamines, an inter-laboratory study was performed by the laboratories from six regulatory agencies with each of the participants using their own analytical procedures to determine the amounts of nitrosamines in a set of identical samples. The results demonstrated that accurate and precise quantitation of trace level nitrosamines can be achieved across multiple analytical procedures and provided insight into the performance characteristics of mass spectrometry-based analytical procedures in terms of accuracy, repeatability and reproducibility.

Inorganic analysis of falsified medical products using X-ray fluorescence spectroscopy and chemometrics.

Falsified medical products are increasingly prevalent on markets, threatening the health of patients. This study describes the benefits of Energy Dispersive X-Ray Fluorescence (ED-XRF) spectroscopy and chemometrics thus highlighting the importance of conducting inorganic analyses on falsified products. The XRF spectrum is a fingerprint containing the contribution of all chemical substances included in a suspect sample's formulation. Multivariate analysis of XRF spectra, using a properly validated classification model, allows for the authentication of suspect samples. The method is rapid, relying on multi-elemental measurements and involving minimal sample preparation. This methodology provided valuable information about samples inorganic composition and enabled the detection of falsifications of several sample types, including medicine, food supplement and cosmetic samples. Five suspect samples of Plavix® were investigated, and their XRF spectra were studied using chemometrics (Principal Component Analysis and Soft Independent Modelling of Class Analogies). A classification model was validated with positive and negative samples, and four suspect samples were identified as being falsified, whilst the fifth was concluded as an authentic medicine. ED-XRF spectroscopy was also applied on another medicine, a food supplement and three cosmetic samples, and high level of zinc was detected in the second sample and mercury was identified in the last. Estimation of the zinc content was possible using the fundamental parameters method. ED-XRF spectroscopy allows the analyst to conclude on the falsification of the samples and then to assess the harm to patient health.

Identification and quantification of ethylene oxide in sterilized medical devices using multiple headspace GC/MS measurement.

This manuscript, based on the ISO 10993-7 approach, describes a multiple HS-GC measurement of residual EO present in sterilized plastic samples. The quantification of EO is done, according to the ISO standard, by addition of EO amounts extracted for each repeated extraction. During the method development, the specificity of the detection of EO regarding acetaldehyde (structural isomer of EO) which may be formed from EO has been ensured and different tests were performed to check a possible influence of the sample preparation. Assays to maximize EO extraction were performed for different materials (Cyclo-olefine Copolymer (COC), Cyclo-olefine Polymer (COP), Silicon, Polyurethane (PUR)) changing extraction temperatures and times for the headspace and the pre-thermal treatment. Results highlight that depending on the material, EO can be more or less retained and thus thermal extraction conditions to maximize the amount of extractible EO from plastics may change accordingly. For COC syringes a validation according to ICH guidelines and an inter-laboratories study were performed. The method has been used for a market survey of EO sterilized medical devices, results obtained are reported in this manuscript.

Raman chemical imaging for spectroscopic screening and direct quantification of falsified drugs.

Falsified drugs are a threat to the health of patients. The analytical control of such products contributes to the fight against this global issue. Raman chemical imaging is a method that relies on consecutive measurements at the surface of a sample, combining spectroscopy, microscopy and chemometrics. This article explores the capabilities of this analytical technique proposing an innovative methodology with spectroscopic screening for the identification of chemical compounds and the direct quantification of the active substance (without prior calibration). Two chemometric methods were used: Multivariate Curve Analysis - Alternate Least Squares for the qualitative analysis and Direct Classical Least Squares for the quantitative analysis. The methodology was optimized with samples prepared in the laboratory and validation parameters were studied. The methodology was then applied to real (authentic and falsified) samples of Viagra® and Plavix®. Despite the presence of fluorescence emission in some samples, the methodology succeeded in the detection of active pharmaceutical ingredients, and in the discrimination of three salts of clopidogrel (in generic formulations of Plavix®). The quantitative deviation from the reference method ranged from -15% to +24% of the active substance content. This deviation may be considered to be acceptable since it is sufficient for assessing the risk to the health of patients and for quickly alerting the health authorities.

Fighting falsified medicines: The analytical approach.

Given the harm to human health, the fight against falsified medicines has become a priority issue that involves numerous actors. Analytical laboratories contribute by performing analyses to chemically characterise falsified samples and assess their hazards for patients. A wide range of techniques can be used to obtain individual information on the organic and inorganic composition, the presence of an active substance or impurities, or the crystalline arrangement of the formulation's compound. After a presentation of these individual techniques, this review puts forward a methodology to combine them. In order to illustrate this approach, examples from the scientific literature (products used for erectile dysfunction treatment, weight loss and malaria) are placed in the centre of the proposed methodology. Combining analytical techniques allows the analyst to conclude on the falsification of a sample, on its compliance in terms of pharmaceutical quality and finally on the safety for patients.

Investigation of the composition of anabolic tablets using near infrared spectroscopy and Raman chemical imaging.

The use of performance enhancing drugs is a widespread phenomenon in professional and leisure sports. A spectroscopic study was carried out on anabolic tablets labelled as 5 mg methandienone tablets provided by police departments. The analytical approach was based on a two-step methodology: a fast analysis of tablets using near infrared (NIR) spectroscopy to assess sample homogeneity based on their global composition, followed by Raman chemical imaging of one sample per NIR profile to obtain information on sample formulation. NIR spectroscopy assisted by a principal components analysis (PCA) enabled fast discrimination of different profiles based on the excipient formulation. Raman hyperspectral imaging and multivariate curve resolution - alternating least square (MCR-ALS) provided chemical images of the distribution of the active substance and excipients within tablets and facilitated identification of the active compounds. The combination of NIR spectroscopy and Raman chemical imaging highlighted dose-to-dose variations and succeeded in the discrimination of four different formulations out of eight similar samples of anabolic tablets. Some samples contained either methandienone or methyltestosterone whereas one sample did not contain an active substance. Other ingredients were sucrose, lactose, starch or talc. Both techniques were fast and non-destructive and therefore can be carried out as exploratory methods prior to destructive screening methods. Copyright © 2015 John Wiley & Sons, Ltd.

HPLC-UV Method for the Identification and Screening of Hydroquinone, Ethers of Hydroquinone and Corticosteroids Possibly Used as Skin-Whitening Agents in Illicit Cosmetic Products.

Corticosteroids, hydroquinone and its ethers are regulated in cosmetics by the Regulation 1223/2009. As corticosteroids are forbidden to be used in cosmetics and cannot be present as contaminants or impurities, an identification of one of these illicit compounds deliberately introduced in these types of cosmetics is enough for market survey control. In order to quickly identify skin-whitening agents present in illegal cosmetics, this article proposes an HPLC-UV method for the identification and screening of hydroquinone, 3 ethers of hydroquinone and 39 corticosteroids that may be found in skin-whitening products. Two elution gradients were developed to separate all compounds. The main solvent gradient (A) allows the separation of 39 compounds among the 43 compounds considered in 50 min. Limits of detection on skin-whitening cosmetics are given. For compounds not separated, a complementary gradient elution (B) using the same solvents is proposed. Between 2004 and 2009, a market survey on "skin-whitening cosmetic" was performed on 150 samples and highlights that more than half of the products tested do not comply with the Cosmetic Regulation 1223/2009 (amending the Council Directive 76/768/EEC).

Identification and quantification of bisphenol A and bisphenol B in polyvinylchloride and polycarbonate medical devices by gas chromatography with mass spectrometry.

A gas chromatography with mass spectrometry method has already been developed and published for the identification and quantification of 14 phthalates and five nonphthalate plasticizers in polyvinylchloride medical devices. In order to assay, in addition to plasticizers, bisphenols A and B possibly present in polyvinylchloride samples, this previous method was extended to the assay of these additional potential endocrine disruptors. Furthermore, as bisphenol A could also be present in polycarbonate samples, the method used for the polyvinylchloride sample was tested and validated for the assay of bisphenols A and B in polycarbonate medical devices. The separation of all compounds, including bisphenols A and B, is obtained on a cross-linked 5%-phenyl/95%-dimethylpolysiloxane capillary column using a temperature gradient. For both plastics, samples are dissolved in tetrahydrofuran followed by a precipitation of the plastic by addition of ethanol. Results obtained point out residual bisphenol A amounts for polycarbonate samples ranging from 0.6 to 0.8% and for polyvinylchloride samples less or equal to 5 ppm. No bisphenol B was detected in the samples tested. For bisphenols A and B, mean recoveries obtained on spiked polyvinylchloride or polycarbonate sample preparations ranged from 87 to 108% in accordance with in-house specification (80-110%).

High-performance liquid chromatography method for the determination of hydrogen peroxide present or released in teeth bleaching kits and hair cosmetic products.

This manuscript presents an HPLC/UV method for the determination of hydrogen peroxide present or released in teeth bleaching products and hair products. The method is based on an oxidation of triphenylphosphine into triphenylphosphine oxide by hydrogen peroxide. Triphenylphosphine oxide formed is quantified by HPLC/UV. Validation data were obtained using the ISO 12787 standard approach, particularly adapted when it is not possible to make reconstituted sample matrices. For comparative purpose, hydrogen peroxide was also determined using ceric sulfate titrimetry for both types of products. For hair products, a cross validation of both ceric titrimetric method and HPLC/UV method using the cosmetic 82/434/EEC directive (official iodometric titration method) was performed. Results obtained for 6 commercialized teeth whitening products and 5 hair products point out similar hydrogen peroxide contain using either the HPLC/UV method or ceric sulfate titrimetric method. For hair products, results were similar to the hydrogen peroxide content using the cosmetic 82/434/EEC directive method and for the HPLC/UV method, mean recoveries obtained on spiked samples, using the ISO 12787 standard, ranges from 100% to 110% with a RSD<3.0%. To assess the analytical method proposed, the HPLC method was used to control 35 teeth bleaching products during a market survey and highlight for 5 products, hydrogen peroxide contents higher than the regulated limit.

Identification and quantification of 14 phthalates and 5 non-phthalate plasticizers in PVC medical devices by GC-MS.

A GC/MS method was developed for the identification and quantification of 14 phthalates: 8 phthalates classified H360 (DBP, DEHP, BBP, DMEP, DnPP, DiPP, DPP and DiBP), 3 phthalates proposed to be forbidden in medical devices (DnOP, DiNP and DiDP) and 3 other phthalates none regulated (DMP, DCHP and DEP) which may interfere with hormone function. In order to identify and quantify other plasticizers that are commonly used in PVC medical devices such as DEHP substitute, 5 non-phthalate plasticizers (ATBC, DEHA, DEHT, TOTM, and DINCH) were included in this study. Analyses are carried out on a GC/MS system with electron impact ionization mode (EI). The separation of plasticizers is obtained on a cross-linked 5%-phenyl/95%-dimethylpolysiloxane capillary column 30m×0.25mm (i.d.)×0.25µm film thickness using a gradient temperature. Compounds quantification is performed by external calibration using an internal standard. Validation elements on standard solutions were determined using the ISO 12787 standard approach. Plasticizers are extracted from PVC medical devices using THF for dissolving the PVC part of the sample followed by precipitation of the PVC by addition of ethanol. The supernatant is injected into a GC/MS system after dilution in ethanol. Different validation elements, including extraction recoveries for all compounds or for DEHP a cross-validation of the extraction process using the European pharmacopoeia monograph 3.1.14 as reference method, are discussed. Results obtained on 61 medical devices in PVC and 12 raw materials used as plasticizers are given.