Quantification Of Active Pharmaceutical Ingredients Research Articles

Qualitative and quantitative analysis of counterfeit fluconazole capsules: A non-invasive approach using NIR spectroscopy and chemometrics

The ultimate goal of the study is to present a method of authentication of hard-shell capsules of medicines packed in polyvinylchloride (PVC) blisters without damaging the primary packaging. This is done by collecting NIR spectra in a non-invasive mode and subsequent analysis of measurements by a one-class classification procedure. The first part of the study demonstrates that NIR spectra collected through a PVC blister and capsule shell do carry information about the medication itself. Firstly, this is done by visual inspection of spectra of the sample, its interfering layers and main pharmaceutical ingredients. Secondly, three regression models for quantification of active pharmaceutical ingredient (API) are built. The possibility of calibration and prediction of API through several nuisance layers using NIR spectroscopy is demonstrated.In order to solve the authentication problem the data driven soft Independent modeling of class analogies method is applied to the collected NIR spectra. The constructed model is validated using laboratory prepared mixtures. Afterwards, the model was applied to real counterfeited samples. Capsules of fluconazole are used for demonstration of the proposed approach."

Downstream processing from hot-melt extrusion towards tablets: A quality by design approach

Since the concept of continuous processing is gaining momentum in pharmaceutical manufacturing, a thorough understanding on how process and formulation parameters can impact the critical quality attributes (CQA) of the end product is more than ever required. This study was designed to screen the influence of process parameters and drug load during HME on both extrudate properties and tableting behaviour of an amorphous solid dispersion formulation using a quality-by-design (QbD) approach. A full factorial experimental design with 19 experiments was used to evaluate the effect of several process variables (barrel temperature: 160–200°C, screw speed: 50–200rpm, throughput: 0.2-0.5kg/h) and drug load (0–20%) as formulation parameter on the hot-melt extrusion (HME) process, extrudate and tablet quality of Soluplus®-Celecoxib amorphous solid dispersions. A prominent impact of the formulation parameter on the CQA of the extrudates (i.e. solid state properties, moisture content, particle size distribution) and tablets (i.e. tabletability, compactibility, fragmentary behaviour, elastic recovery) was discovered. The resistance of the polymer matrix to thermo-mechanical stress during HME was confirmed throughout the experimental design space. In addition, the suitability of Raman spectroscopy as verification method for the active pharmaceutical ingredient (API) concentration in solid dispersions was evaluated. Incorporation of the Raman spectroscopy data in a PLS model enabled API quantification in the extrudate powders with none of the DOE-experiments resulting in extrudates with a CEL content deviating>3% of the label claim. This research paper emphasized that HME is a robust process throughout the experimental design space for obtaining amorphous glassy solutions and for tabletting of such formulations since only minimal impact of the process parameters was detected on the extrudate and tablet properties. However, the quality of extrudates and tablets can be optimized by adjusting specific formulations parameters (e.g. drug load).

Study of the matrix effect on the PIXE quantification of active pharmaceutical ingredients in different formulations

While Particle Induced X-ray Emission technique (PIXE) is an accurate technique to quantify Active Pharmaceutical Ingredients (API’s) via the analysis of their heteroatoms, each drug (formulation) may require a specific quantification procedure due to its distinct matrix composition.The commercial Fludinium® drug, which has two active ingredients Clidinium Bromide (C22H26NO3Br) and Dihydrochloride Trifluoperazine (C21H24N3F3S. 2HCl) has been taken as a case study in this work. Different amounts of its API’s and its placebo were mixed to provide various formulations.The matrix effect on the quantification of the three heteroatoms (chlorine, sulfur and bromine) related to the above API’s in different formulations has been studied. In fact, chlorine which is in its hydrochloride form in the API was not eventually considered for calculation due to its instability under beam. The calculation of bromine amount via its Kα or its Lα rays was found to be independent of the matrix composition and it was rapidly done by simple comparison to an external standard. However, the calculation of sulfur, via its Kα, was highly dependent on the matrix composition. Therefore, to achieve an accurate quantification a more sophisticated calculation method was used by means of the GUPIX code.

Raman spectroscopy and capillary zone electrophoresis for the analysis of degradation processes in commercial effervescent tablets containing acetylsalicylic acid and ascorbic acid

In order to ensure the stability of pharmaceutical products appropriate manufacturing and storage conditions are required. In general, the degradation of active pharmaceutical ingredients (APIs) and subsequent formation of degradation products affect the pharmaceutical quality. Thus, a fast and effective detection and characterization of these substances is mandatory. Here, the applicability of Raman spectroscopy and CZE for the characterization of the degradation of effervescent tablets containing acetylsalicylic acid (ASA) and ascorbic acid (AA) was evaluated. Therefore, a degradation study was performed analyzing tablets from two different manufacturers at varying conditions (relative humidity (RH) 33%, 52% and 75% at 30°C). Raman spectroscopy combined with principal component analysis could be successfully applied for the fast and easy discrimination of non-degraded and degraded effervescent tablets after a storage period of approximately 24h (RH 52%). Nevertheless, a clear identification or quantification of APIs and degradation products within the analyzed tablets was not possible, i.a. due to missing reference materials. CZE-UV enabled the quantification of the APIs (ASA, AA) and related degradation products (salicylic acid (SA); semi-quantitative also mono- and diacetylated AA) within the complex tablet mixtures. The higher the RH, the faster the degradation of ASA and AA as well as the formation of the degradation products. Mono- and diacetylated AA are major primary degradation products of AA for the applied effervescent tablets. A significant degradation of the APIs was detected earlier by CZE (6–12h, RH 52%) than by Raman spectroscopy. Summarized, Raman spectroscopy is well-suited as quick test to detect degradation of these tablets and CZE can be utilized for further detailed characterization and quantification of specific APIs and related degradation products.

Capillary-induced Homogenization of Matrix in Paper: A Powerful Approach for the Quantification of Active Pharmaceutical Ingredients Using Mass Spectrometry Imaging.

Herein we present a novel approach for the quantification of active pharmaceutical ingredients (APIs) using mass spectrometry imaging. This strategy uses a filter paper previously “eluted” with a MALDI matrix solution as a support for analyte application. Samples are submitted to mass spectrometry imaging (MSI) and quantification through characteristic fingerprints is ultimately performed. Results for the content of rosuvastatin from a known formulation are comparable to those obtained with a validated HPLC method.

Quantitative analysis of topical gels and ointments by FT-Raman spectroscopy

A method for quantitative determination of ibuprofen (IBU), naproxen (NAP), methyl salicylate (MES) and menthol (MNT) in commercial topical gels and ointments using partial least squares (PLS) models based on FT-Raman spectra is described. The calculated relative standard errors of prediction (RSEP) were found to be in the range of 2.1–3.2% for the calibration and validation data sets. Two commercial topical gels containing 5.0% of IBU and 10% of NAP (w/w), as well as one ointment containing 15% of MES and 10% of MNT (w/w) as active pharmaceutical ingredients (APIs), were successfully quantified using the developed models with recoveries in the 99.2–101.5% range. The proposed procedure can be used as a fast, reliable and economic method for the quantification of APIs in topical gels and ointments.

Post-marketing surveillance of anti-malarial medicines used in Malawi.

BackgroundThe growing concern over the extent of anti-malarial medicine resistance in sub-Saharan Africa, driven largely by administration of sub-therapeutic doses derived from falsified and substandard medicines necessitates regular monitoring of the quality of these medicines to avert any potential public health disaster. This study aimed at determining the active pharmaceutical ingredient (API) content of anti-malarial medicines available in Malawi with respect to the manufacturers’ label claim and pharmacopoeia specifications.MethodsSamples of anti-malarial medicines (112) collected from both licensed and unlicensed markets throughout Malawi were subjected to visual inspection of dosage form and packaging, and registration verification with the regulatory body. Basic (colourimetric) tests were employed to establish the presence and identity of the requisite APIs. Semi-quantitative thin layer chromatography (SQ-TLC) was employed as a quick assay for the verification of identity and estimation of the API content while HPLC assays were used to quantify the APIs. The results were compared with pharmacopoeia specifications and manufacturers’ label claims. For combination therapies, a sample was considered to have failed if one or more of its component APIs did not meet pharmacopoeia specifications.ResultsThere was 86.6% registration status and 100% compliance with visual inspection and basic tests confirming the presence of requisite APIs. The identification test was confirmed by the SQ-TLC assay. API quantification by HPLC assay however, showed that 88.4% (99/112) of the samples failed the quality tests due to the presence of either insufficient or excessive API.ConclusionsThe results suggest the existence of substandard anti-malarial medicines in Malawi. The presence of both excessive and insufficient artemisinin-based and non-artemisinin-based API, clearly points to poor adherence to GMP and improper handling during storage or distribution. The country relies heavily on imported anti-malarial medicines so there is an urgent need to carry out regular and thorough post-market surveillance of medicines to ensure better quality health care delivery.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-015-0637-z) contains supplementary material, which is available to authorized users.

Development and validation of a stability indicating HPLC method for the simultaneous analysis of lopinavir and ritonavir in fixed-dose combination tablets

Abstract Objectives A simple, accurate, precise and rapid stability indicating HPLC method for simultaneous determination of Lopinavir and Ritonavir in combined dosage forms. Methods A validated stability indicating reversed phase high-performance liquid chromatographic method was developed for the quantitative determination of two antiviral drugs viz. lopinavir (LPV) and ritonavir (RTV) on Agilent TC C18 (2) 250 × 4.6 mm, 5 μ column using mobile phase composition of acetonitrile: 0.05 M phosphoric acid (55: 45, v/v) at a flow rate of 1.2 ml/min. Results Quantification was achieved with ultraviolet detection at 240 nm. The retention time obtained for ritonavir was at 4.35 min and for lopinavir was at 6.68 min. The result obtained with the detector response was found to be linear in the concentration range of 8–48 μg/ml for lopinavir and 2–12 μg/ml for ritonavir. This method has been validated and shown to be specific, sensitive, precise, linear, accurate, rugged, robust and fast. LPV and RTV were subjected to different accelerated stress conditions. The degradation products, were well resolved from the pure drug with significantly different retention time values. Conclusion It is concluded that this method can be applied for routine quality control of LPV and RTV in tablet dosage forms as well as in bulk drug. Hence the proposed method is highly sensitive, precise and accurate and it successfully applied for the reliable quantification of active pharmaceutical ingredient (API) content in the commercial formulations of lopinavir and ritonavir.

Quantification of active pharmaceutical ingredient and impurities in sildenafil citrate obtained from the Internet.

The accessibility of prescription drugs produced outside of the United States, most notably sildenafil citrate (innovator product, Viagra®), has been made much easier by the Internet. Of greatest concern to clinicians and policymakers is product quality and patient safety. The US Food and Drug Administration (FDA) has issued warnings to potential buyers that the safety of drugs purchased from the Internet cannot be guaranteed, and may present a health risk to consumers from substandard products. The objective of this study was to determine whether generic sildenafil citrate tablets from international markets obtained via the Internet are equivalent to the US innovator product regarding major aspects of pharmaceutical quality: potency, accuracy of labeling, and presence and level of impurities. This will help identify aspects of drug quality that may impact public health risks. A total of 15 sildenafil citrate tablets were obtained for pharmaceutical analysis: 14 generic samples from international Internet pharmacy websites and the US innovator product. According to US Pharmacopeial guidelines, tablet samples were tested using high-performance liquid chromatography for potency of active pharmaceutical ingredient (API) and levels of impurities (impurities A, B, C, and D). Impurity levels were compared with International Conference on Harmonisation (ICH) limits. Among the 15 samples, 4 samples possessed higher impurity B levels than the ICH qualification threshold, 8 samples possessed higher impurity C levels than the ICH qualification threshold, and 4 samples possessed more than 1% impurity quantity of maximum daily dose (MDD). For API, 6 of the samples failed to fall within the 5% assay limit. Quality assurance tests are often used to detect formulation defects of drug products during the manufacturing and/or storage process. Results suggest that manufacturing standards for sildenafil citrate generic drug products compared with the US innovator product are not equivalent with regards to potency and levels of impurities. These findings have implications for safety and effectiveness that should be addressed by clinicians to safeguard consumers who choose to purchase sildenafil citrate and foreign-manufactured drugs, in general, via the Internet.

Method Development and Validation for Simultaneous Determination of Ezetimibe and Simvastatin In Combined Pharmaceutical Dosage Form By RP-HPLC Method

A simple, rapid reverse phase high-performance liquid chromatographic method was developed and validated for the simultaneous estimation of Ezetimibe and Simvastatin in bulk and pharmaceutical dosage forms. Chromatography was carried out by using Chromosil C-18,column having 250 x 4.6mm internal diameter with a mixture of Methanol:Acetonitrile:0.1%Orthophosphoric Aid in the ratio of 75:20:05 (v/v/v) as mobile phase. Determination of the different analytical parameters such as linearity, precision, accuracy, and specificity, limit of detection (LOD) and limit of quantification (LOQ) was done. The calibration curve was found to be linear for each analyte in the desired concentration range. The average recovery was found to be 99.88 and 100.12 for Ezetimibe and Simvastatin respectively. The proposed method is highly sensitive, precise and accurate, which was evident from the LOD value of 1.2ppm and 0.25ppm for Ezetimibe and Simvastatin respectively and hence the present method can be applied successfully for the quantification of active pharmaceutical ingredient (API) content in the combined formulations of Ezetimibe and Simvastatin. DOI: http://dx.doi.org/10.3329/ijpls.v2i2.15450 International Journal of Pharmaceutical and Life Sciences Vol.2(2) 2013: 60-69

A quantitative method using near infrared imaging spectroscopy for determination of surface composition of tablet dosage forms: an example of spirolactone tablets

In this work, near infrared (NIR) imaging spectroscopy was employed in the study of the distribution of the active pharmaceutical ingredient (API) spironolactone and its excipients in tablets. Analyses were performed using 50 µm spatial resolution to analyze 16 mm² of each standard tablet. Interval partial least squares models were used for API and excipients quantification in every pixel in order to obtain concentration maps for each compound. It was obtained errors of quantification between 0.49 and 1.26% when performed the cross-validation using spectral average of each tablet. These calibration models were used to predict individual concentration of each compound in the tablet, in every pixel. The average concentration of all pixels, for each compound yield errors, was between 0.05 and 1.06%. These results indicate that the models were able to quantify all compounds in each pixel. This approach is necessary since it is not possible to know the real concentration of each compound in the pixels.

Thin-layer Chromatography–Nuclear Magnetic Resonance Spectroscopy – A Versatile Tool for Pharmaceutical and Natural Products Analysis

Thin-layer chromatography (TLC) is a mature and very established technique, frequently used in many fields of applications ranging from natural product analysis to chemical or pharmaceutical applications. The introduction of a commercially available TLC-MS interface was a major step complementing the ease of use of TLC with structural elucidation power of mass spectrometry (MS). The TLC-MS interface simplifies the workflow dramatically to gain structural information directly from TLC separations. This article describes the potential of TLC-nuclear magnetic resonance spectroscopy (NMR) utilizing the TLC-MS interface to straightforwardly characterize zones of interest by NMR spectroscopy with a focus on quantification of active pharmaceutical ingredients (API) in formulations and identification of active principles in plant extracts.

DEVELOPMENT AND VALIDATION OF RP-HPLC AND ULTRAVIOLET SPECTROPHOTOMETRIC METHODS OF ANALYSIS FOR SIMULTANEOUS DETERMINATION OF PARACETAMOL AND LORNOXICAM IN PHARMACEUTICAL DOSAGE FORMS

A high performance liquid chromatographic and an UV spectrophotometric method were developed and validated for the simultaneous determination of paracetamol and lornoxicam in pharmaceutical formulations. The different analytical performance parameters such as linearity, precision, accuracy, specificity, limit of detection (LOD), and limit of quantification (LOQ) were determined according to International Conference on Harmonization (ICH) guidelines. HPLC was carried out by isocratic method on a reversed-phase C18 column and potassium dihydrogen phosphate buffer (pH adjusted to 3.5): acetonitrile (55:45, v/v) as eluent, at a flow rate of 1.0 mL/min. The UV spectrophotometric determinations were performed at 273 and 405 nm for paracetamol and lornoxicam, respectively. The linearity of the calibration curves for each analyte in the desired concentration range is good (r2 > 0.9997) by both the HPLC and UV methods. Both the methods were accurate and precise with recoveries in the range of 98 and 101% for the drugs and relative standard deviation (RSD) <5%. Moreover, the accuracy and precision obtained with HPLC correlated well with the UV method. However, UV spectroscopy can be an inexpensive, reliable, and less time consuming alternative for chromatographic analysis. The proposed methods are highly sensitive, precise, and accurate; hence, they were successfully applied for the reliable quantification of Active Pharmaceutical Ingredient (API) content in the commercial formulations of paracetamol and lornoxicam. Though the UV method is more practicable, the HPLC method shows application in stability indicating studies in addition to their assay.

Raman spectroscopy for the in-line polymer–drug quantification and solid state characterization during a pharmaceutical hot-melt extrusion process

The aim of this study was to evaluate the suitability of Raman spectroscopy as a Process Analytical Technology (PAT) tool for the in-line determination of the active pharmaceutical ingredient (API) concentration and the polymer–drug solid state during a pharmaceutical hot-melt extrusion process. For in-line API quantification, different metoprolol tartrate (MPT) – Eudragit® RL PO mixtures, containing 10%, 20%, 30% and 40% MPT, respectively, were extruded and monitored in-line in the die using Raman spectroscopy. A PLS model, regressing the MPT concentrations versus the in-line collected Raman spectra, was developed and validated, allowing real-time API concentration determination. The correlation between the predicted and real MPT concentrations of the validation samples is acceptable ( R 2 = 0.997). The predictive performance of the calibration model is rated by the root mean square error of prediction (RMSEP), which is 0.59%. Two different polymer–drug mixtures were prepared to evaluate the suitability of Raman spectroscopy for in-line polymer–drug solid state characterization. Mixture 1 contained 90% Eudragit® RS PO and 10% MPT and was extruded at 140 °C, hence producing a solid solution. Mixture 2 contained 60% Eudragit® RS PO and 40% MPT and was extruded at 105 °C, producing a solid dispersion. The Raman spectra collected during these extrusion processes provided two main observations. First, the MPT Raman peaks in the solid solution broadened compared to the corresponding solid dispersion peaks, indicating the presence of amorphous MPT. Second, peak shifts appeared in the spectra of the solid dispersion and solid solution compared to the physical mixtures, suggesting interactions between Eudragit® RS PO and MPT, most likely hydrogen bonds. These shifts were larger in the spectra of the solid solution. DSC analysis confirmed these Raman solid state observations and the interactions seen in the spectra. Raman spectroscopy is a potential PAT-tool for in-line determination of the API concentration and the polymer–drug solid state during pharmaceutical hot-melt extrusion.

Acoustic-resonance spectrometry as a process analytical technology for the quantification of active pharmaceutical ingredient in semi-solids.

The purpose of this study was to demonstrate acoustic resonance spectrometry (ARS) as an alternative process analytical technology to near infrared (NIR) spectroscopy for the quantification of active pharmaceutical ingradient (API) in semi-solids such as creams, gels, ointments, and lotions. The ARS used for this research was an inexpensive instrument constructed from readily available parts. Acoustic-resonance spectra were collected with a frequency spectrum from 0 to 22.05 KHz. NIR data were collected from 1100 to 2500 nm. Using 1-point net analyte signal (NAS) calibration, NIR for the API (colloidal oatmeal [CO]) gave anr 2 prediction accuracy of 0.971, and a standard error of performance (SEP) of 0.517%CO. ARS for the API resulted in anr 2 of 0.983 and SEP of 0.317%CO. NAS calibration is compared with principal component regression. This research demonstrates that ARS can sometimes outperform NIR spectrometry and can be an effective analytical method for the quantification of API in semi-solids. ARS requires no sample preparation, provides larger penetration depths into lotions than optical techniques, and measures API concentrations faster and more accurately. These results suggest that ARS is a useful process analytical technology (PAT).

Quantitative determination of diclofenac sodium and aminophylline in injection solutions by FT-Raman spectroscopy

The FT-Raman quantification of diclofenac sodium and aminophylline commercial injection solutions was performed. The efficiency of various spectra treatment procedures including classical univariate intensity ratio and multivariate partial least squares (PLS) and principal component regression (PCR) methods was compared. First, the calibration models were built using unnormalised spectra. Next, spectra normalised by the intensity of a selected band of CH 3CN added as an internal standard to the studied samples were utilised. To compare the predictive ability of the models constructed, the relative standard error of prediction (RSEP) was calculated. The errors found for multivariate calibrations were a few times smaller than those for the univariate ones. Usually, the most effective was the PLS method, for which RSEP values of the order of 1–2% for calibration and 2–3% for testing data sets were obtained. Four commercial preparations of diclofenac sodium and one of aminophylline containing by weight, 2.4% of the active pharmaceutical ingredient (API) were quantified applying the developed models. Concentrations found from the Raman data analysis correlate with the declared values and the results of reference analyses. For the studied diclofenac sodium solutions they amount to 99.2–101.2% of the former and 101.2–102.4% of the latter quantities for the PLS models optimised for each medicine based on unnormalised spectra. These values for the aminophylline preparation were found to be 101.0 and 99.1%, respectively. It shows that the proposed procedure based on the chemometric treatment of FT-Raman spectra can be a fast and convenient alternative to the standard pharmacopoeial procedures of API quantification even in relatively diluted injection solutions.