Analysis Of Impurities In Pharmaceuticals Research Articles

Pharmaceutical analysis by NMR can accommodate strict impurity thresholds: The case of choline

The ICH guidelines recommend reporting thresholds for regular impurities in drug substances at the level of 0.05% or 0.03% (w/w) depending on the maximum daily intake. Therefore, any instrumental method of analysis applicable to the impurity analysis should be able to detect and quantify the analytes at those levels. This investigation was designed to verify the suitability of 1H NMR spectroscopy for the detection of impurities, as a first step in the process before attempting quantification. In order to minimize demand on equipment, this study employed a 400 MHz instrument for structural confirmation and signal assignments of choline (1) and O-(2-hydroxyethyl)choline (2), a known impurity. The limit of detection (LOD) of 2 in 10 mg of 1 was established as 0.01% on a 400 MHz instrument and 2% on a 60 MHz (benchtop) NMR spectrometer. Thus, impurities for which quantification is required are readily detected at 400 MHz or above. These results are in contrast to the widespread belief that 1H NMR sensitivity is insufficient for pharmaceutical impurity analysis. The choice of solvent was recognized as a critical parameter for 1H NMR LOD analysis. Furthermore, publicly available NMR raw data (HMDB) proved to be valuable for unveiling the otherwise cryptic information hidden in complex signal patterns via 1H NMR iterative Full Spin Analysis. Finally, the study uncovered the less noticed, yet characteristic, 14N–1H coupling in the -N+(CH3)3 groups, adding strong arguments for the Raw NMR Data Initiative. Collectively, the data prove that the analytical capabilities of high-field NMR easily fulfill the ICH requirements for detection of impurity in the presence of an actual substance of interest which makes it a step closer to achieving regulatory standards.

Pharmaceutical impurity analysis by comprehensive two-dimensional temperature responsive × reversed phase liquid chromatography

In this study, the possibilities of temperature responsive × reversed phase liquid chromatography (TRLC × RPLC) are assessed in terms of pharmaceutical impurity analysis. Due to the increased peak capacity per unit time they offer, two-dimensional LC approaches are gaining relevance for the analysis of complex drug formulations. Because the latter depicts a larger predisposition for the occurrence of an increased number of impurities, current 1D-HPLC approaches often prove insufficient. Since many LC × LC methods are limited by modulation, solvent compatibility, orthogonality, and sensitivity issues, the combination of TRLC × RPLC is explored in this work for pharmaceutical impurity analysis. As this combination of a purely aqueous separation with RPLC allows for systematic and optimization-free refocusing in the second dimension, it opens possibilities for generic LC × LC requiring minimal to no method development, in this way overcoming a major perceived contemporary hurdle of LC × LC. The approach is demonstrated with a representative mixture of 17 solutes comprising 11 corticosteroids and 6 progestogens. Orthogonality and peak capacities were assessed on three RP core-shell column selectivities (Poroshell EC-C18, phenyl-hexyl and PFP). Although the TRLC × EC-C18 combination offered somewhat better orthogonality, the combination with the PFP column proved the best for the separation at hand. Depending on the composition of the mixture, the use of full, shifted, or segmented gradients allowed facile optimization of the separation. The developed platform allowed detection of the impurities at the 0.05% level compared to a selected main compound, while also opening up possibilities for analysis of formulations comprising two active ingredients.

Development of an achiral-chiral 2-dimensional heart-cutting platform for enhanced pharmaceutical impurity analysis.

A new broadly applicable achiral-chiral 2-dimensional heart-cutting (LC-LC) platform is designed comprising a multi-column selection approach in the chiral dimension. As both dimensions are operated in a highly aqueous reversed-phase type mode, analysis of a broad range of pharmaceutical solutes (LogP: 1.49-5.7) and their impurities becomes possible, while breakthrough issues arising due to solvent immiscibility and peak broadening phenomena in the second dimension are practically circumvented. These aspects, together with the chromatographic and quantitative performances are systematically assessed for various transfer loop sizes (50, 100, 200 and 500µl), column diameters (2.0 and 4.6mm), and various gradients (10, 20 and 40min) with a mixture of five racemates covering a broad range in polarity. In order to broaden the selectivity of the second dimension, an automated chiral screening is performed comprising six chiral columns, allowing baseline separation for all enantiomers and a chiral resolution up to 17.21 for some of the racemates. The performance of the platform is also assessed in pharmaceutical drug development samples. A hybrid high-resolution (HiRes) sample approach is thereby used, which proves to be effective for precise confirmation of the relative prevalence of the impurities compared to the principal compound in all studied cases. The co-eluted impurities were thereby effectively detected and quantified down to the 0.05% level. The obtained figures of merits indicate the suitability of the platform for implementation in industry.

Analysis of impurities in pharmaceuticals by LC-MS with cold electron ionization.

Pharmaceuticals require careful and precise determination of their impurities that might harm the user upon consumption. Although today, the most common technique for impurities identification is liquid chromatography-mass spectrometry (LC-MS/MS), it has several downsides due to the nature of the ionization method. Also, the analyses in many cases are targeted thus despite being present, some of the compounds will not be revealed. In this paper, we propose and show a new method for untargeted analysis and identification of impurities in active pharmaceutical ingredients (APIs). The instrument used for these analyses is a novel electron ionization (EI) LC-MS with supersonic molecular beams (SMB). The EI-LC-MS-SMB was implemented for analyses of several drug samples spiked with an impurity. The instrument provides EI mass spectra with enhanced molecular ions, named Cold EI, which increases the identification probabilities when the compound is identified with the aid of an EI library like National Institute of Standards and Technology (NIST). We analyzed ibuprofen and its impurities, and both the API and the expected impurity were identified with names and structures by the NIST library. Moreover, other unexpected impurities were found and identified proving the ability of the EI-LC-MS-SMB system for truly untargeted analysis. The results show a broad dynamic range of four orders of magnitude at the same run with a signal-to-noise ratio of over 10 000 for the API and almost uniform response.

The use of glutathione as a derivatising agent for the analysis of Michael acceptor genotoxic impurities in pharmaceuticals

This work describes the development strategy and validation of an analytical method to determine trace levels of a Michael acceptor containing genotoxic impurity in a pharmaceutical intermediate.

Extending (Q)SARs to incorporate proprietary knowledge for regulatory purposes: A case study using aromatic amine mutagenicity

Statistical-based and expert rule-based models built using public domain mutagenicity knowledge and data are routinely used for computational (Q)SAR assessments of pharmaceutical impurities in line with the approach recommended in the ICH M7 guideline. Knowledge from proprietary corporate mutagenicity databases could be used to increase the predictive performance for selected chemical classes as well as expand the applicability domain of these (Q)SAR models. This paper outlines a mechanism for sharing knowledge without the release of proprietary data. Primary aromatic amine mutagenicity was selected as a case study because this chemical class is often encountered in pharmaceutical impurity analysis and mutagenicity of aromatic amines is currently difficult to predict. As part of this analysis, a series of aromatic amine substructures were defined and the number of mutagenic and non-mutagenic examples for each chemical substructure calculated across a series of public and proprietary mutagenicity databases. This information was pooled across all sources to identify structural classes that activate or deactivate aromatic amine mutagenicity. This structure activity knowledge, in combination with newly released primary aromatic amine data, was incorporated into Leadscope's expert rule-based and statistical-based (Q)SAR models where increased predictive performance was demonstrated.

Use of a Field-Portable XRF Instrument To Facilitate Metal Catalyst Scavenger Screening

The analysis of elemental impurities in pharmaceuticals has traditionally been performed using plasma-based instrumental techniques, such as inductively coupled plasma-optical emission spectroscopy or inductively coupled plasma-mass spectrometry, due to their inherent sensitivity, selectivity, and precision; however, during development activities, there are a variety of sample types that do not require the sensitivity that these expensive and time-consuming techniques provide. Pharmaceutical process chemists often need a rapid and relative determination of the elemental content to direct them in optimizing the effectiveness or efficiency of their synthetic process. This article will provide the application of a field-portable X-ray fluorescence instrument as a hands-on screening tool for chemists to determine the effectiveness of metal catalyst scavenger studies. Three case studies will be provided to illustrate the usefulness of the technique.

Identification, control strategies, and analytical approaches for the determination of potential genotoxic impurities in pharmaceuticals: a comprehensive review.

Potential genotoxic impurities in pharmaceuticals at trace levels are of increasing concern to both pharmaceutical industries and regulatory agencies due to their possibility for human carcinogenesis. Molecular functional groups that render starting materials and synthetic intermediates as reactive building blocks for small molecules may also be responsible for their genotoxicity. Determination of these genotoxic impurities at trace levels requires highly sensitive and selective analytical methodologies, which poses tremendous challenges on analytical communities in pharmaceutical research and development. Experimental guidance for the analytical determination of some important classes of genotoxic impurities is still unavailable in the literature. Therefore, the present review explores the structural alerts of commonly encountered potential genotoxic impurities, draft guidance of various regulatory authorities in order to control the level of impurities in drug substances and to assess their toxicity. This review also describes the analytical considerations for the determination of potential genotoxic impurities at trace levels and finally few case studies are also discussed for the determination of some important classes of potential genotoxic impurities. It is the authors' intention to provide a complete strategy that helps analytical scientists for the analysis of such potential genotoxic impurities in pharmaceuticals.

Supercritical fluid chromatography and its application to analysis and preparation of high-purity compounds

The principles of supercritical fluid chromatography (SFC) and its application in various fields are reviewed. A comparison of high-performance liquid chromatography (HPLC) and SFC is performed. In discussing the applications, emphasis is placed on the possibility of using SFC in analysis of impurities in pharmaceuticals and biologically important substances and in preparation of high-purity substances. In describing detectors for SFC, special attention is paid to mass-spectrometric detectors.

Development of an orthogonal method for mometasone furoate impurity analysis using supercritical fluid chromatography

While supercritical fluid chromatography (SFC) has received great popularity in chiral separation and purification, it has rarely been used for trace level pharmaceutical impurity analysis, partially due to the limitation of instrument sensitivity. In this study, a packed column SFC method has been developed for the quantitative analysis of mometasone furoate and its trace level impurities. The UV detection was optimized to improve the sensitivity by 2–4 fold. In combination with an increased sample concentration, this SFC method is capable of trace level (0.05% of the active) analysis of the impurities. The SFC method used a silica column and a mobile phase consisting of CO 2 and methanol. The new method provides an orthogonal selectivity complementary to the reversed phase HPLC (RP-HPLC) method. All of the impurities and the active were baseline separated within 12 min on SFC, which is less than one third of the RP-HPLC method run time. The method was also partially validated for linearity, accuracy, precision (repeatability), and limit of quantitation. This study demonstrated that the SFC method, with improved sensitivity, can be a valuable tool to provide orthogonal selectivity for trace level impurity separation. With further validation, the method may be suitable for release testing and stability testing for mometasone furoate drug substance.

Analysis of potential genotoxic impurities in pharmaceuticals by two-dimensional gas chromatography with Deans switching and independent column temperature control using a low-thermal-mass oven module

The analysis of potential genotoxic impurities (PGIs) in active pharmaceutical ingredients (APIs) is a challenging task. The target limit of detection for a PGI in an API is typically 1 ppm (1 microg/g API). This is about 500 times lower than for classical impurity analysis. Consequently, analytical methods for trace analysis, mostly in combination with MS detection, need to be applied for the qualitative and quantitative determination of these impurities. A two-dimensional capillary GC method is presented that can be used for the determination of some target PGIs. A concentrated solution of the API sample is directly introduced in the GC-MS system, using an apolar column for first-dimension separation. The fraction (heart-cut) containing the PGIs is transferred to a second capillary column, installed in a low-thermal-mass oven (LTM). The LTM focuses the heart-cut(s) and allows independent temperature-programmed analysis with a polar second-dimension column. The API, solvent, and derivatization agents are not introduced in the second column or in the MS detector, avoiding contamination, column degradation, and target analyte peak detection/integration issues. The performance of this set-up is illustrated by the analysis of some Michael-reactive acceptor PGIs and haloalcohols in carbamazepine as test matrix. Excellent reproducibility (<10% RSD) at the low parts per million level and low detection limits (<1 ppm) were obtained.

A comparison study of sample dissolution media in headspace analysis of organic volatile impurities in pharmaceuticals

Abstract Four sample dissolution media, N , N -dimethylacetamide (DMA), N , N -dimethyl-formamide (DMF), 1,3-dimethyl-2-imidazolidinone (DMI) and water were studied and compared in gas chromatographic headspace analysis of organic volatile impurities (OVIs) in drug substances. Emphasis was placed on fundamental parameters of headspace analysis such as maximum equilibrium temperature, precision, sensitivity and accuracy. The impacts of equilibrium time, temperature and salting out effect were determined and discussed in detail. It is found that the partition of an analyte in the sample solution influence not only the sensitivity but also the precision and accuracy of the analysis. Although the boiling point of DMI was much higher than those of DMF and DMA, they show the same maximum equilibrium temperature (110 °C). Salting out effect in aqueous sample solution diminishes the matrix effect of headspace analysis and increases the sensitivity of polar analytes. It is concluded that: (1) Water and DMI are the best hydrophilic and hydrophobic medium, respectively; (2) Anti-polarity effect is an approach to enhance the sensitivity; (3) Polar and high boiling point OVIs tend to involve problems of analysis precision, sensitivity and accuracy.