Analytical Method Development Articles
A simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and validated for the estimation of telmisartan and hydrochlorothiazide simultaneously in combined dosage forms. The stationary phase used was precoated silica gel 60F254. The mobile phase used was a mixture of chloroform: methanol: toluene (2:5:5 v/v/v). The detection of spots was carried out at 272 nm. The method was validated in terms of linearity, accuracy, precision and specificity.
A new reversed-phase high performance liquid chromatography method was developed and validated for the simultaneous determination of losartan potassium and atenolol in tablets.
Two simple and sensitive visible spectrophotometric methods (A and B) have been developed for the quantitative estimation of nitazoxanide, in bulk drug and pharmaceutical dosage forms. Methods were based on the formation of reddish yellow coloured and green coloured chromogens, which were measured at 544 nm and 715 nm, respectively. The results obtained with the proposed methods are in good agreement with the labelled amounts when tablet dosage forms were analysed.
A simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and validated for the determination of cefuroxime axetil in dosage form. The stationary phase used was precoated silica gel 60F 254 . The mobile phase used was a mixture of chloroform:methanol:toluene (4:2:2 v/v). The detection of spot was carried out at 290 nm. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found to be linear between 300 to 800 ng/spot for cefuroxime axetil.
Valdecoxib is a nonsteroidal antiinflammatory drug, and it is listed in class 2 of biopharmaceutic classification of drugs. Valdecoxib is a poorly water-soluble and highly permeable drug. In the present study a new dissolution medium was developed, as there is no official dissolution medium available in the literature. The composition of the dissolution medium was selected on the basis of solubility data at 37°. Solubility data revealed that addition of surfactant may be suitable as dissolution medium.
A simple, rapid, selective and sensitive reversed phase HPLC method was developed and validated for the determination of rebamipide from plasma. The drug was extracted with a mixture of chloroform and isopropyl alcohol. Rebamipide was measured in plasma using a validated HPLC method with UV detection at 280 nm. Chromatographic peaks were separated on a 5 µm C-18 silica column using a mixture of acetonitrile, water, methanol and acetic acid as a mobile phase.
Several gaps in current regulatory guidelines that govern the analytical method life cycle for the testing of biopharmaceuticals are identified. Strategic guidance on how to monitor and control the life cycle of an analytical test method is provided in this article. Analytical method transfer, analytical method component equivalency, and analytical method comparability protocols are discussed in light of risk-based strategies for validation extensions. The use of an analytical method maintenance program is suggested to control over time the predictable risk to patients and firm.
In the first part of this article1 we discussed the GAMP Good Practice Guide (GPG) for the Validation of Laboratory Computerized Systems.2 We looked at the advantages offered by the System Implementation Life Cycle (SILC) in contrast to the complexity of the system classification proposed in the GPG.
In this part I'll look at the risk assessment methodology, the new US Pharmacopeia (USP) general chapter <1058>,3 which is based upon the AAPS analytical equipment qualification white paper,4 and suggest a way forward to unite the qualification of equipment with the validation of the controlling laboratory computers.
Risk Assessment Methodology
Figure 1: GAMP GPG risk management process.
The design of pharmaceutical water systems has always been part science and part alchemy, and unfortunately, it is not likely to change in the near future. Daily rhetoric is rife with statements from informed and intelligent people alleging design requirements that do not actually exist.
The problem is that if you do not have documentation from a reliable source that supports or contradicts these statements, they are nearly impossible to refute. As a result, they are often incorporated into system design.
An isocratic HPLC method for the determination of phenol and nitrophenols (4-nitrophenol, 2-nitrophenol, 4,6-dinitro-o-cresol and 2,4-dinitrophenol) has been developed and validated using 2-chlorophenol as internal standard (IS) and a monolithic column in tap water samples. Prior to HPLC, the method requires solid-phase extraction (SPE) using polymeric Lichrolut EN cartridges. The method development involved the study of methanol and acetonitrile as organic modifiers, pH and flow-rate using a Chromolith RP-18e (150 mm × 4.6 mm I.D.) column. After comparing the performance of the separations obtained with both organic modifiers, the optimum separation of these compounds was achieved using 50 mM acetate buffer (pH 5.0)-acetonitrile (80:20, v/v) as mobile phase, 3 mL min-1 flow-rate and UV detection at maximum absorbance wavelength.
Purpose. A sensitive, robust, and selective liquid chromatographic – tandem mass spectrometric method (LC-MS/MS) was developed and validated for paroxetine quantification in human EDTA plasma. Methods. Sample preparation was based on liquid-liquid extraction using a mixture of ethyl acetate/hexane (50/50; v/v) to extract the drug and internal standard from plasma. Chromatography was performed on a C-18 analytical column and the retention times were 1.6 and 1.7 for paroxetine and fluoxetine (IS), respectively.
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Reproducibility? Ruggedness? Robustness? To many people, all of these terms mean the same thing. But in reality, in the words of a popular children's program, "one of these things is not like the other." An earlier "Validation Viewpoint" column (1) touched briefly on this topic, but this column will explore the topic in a little more detail. So, for the purposes of this discussion, the "R-word" we are most interested in is robustness. However, first we need a few clarifications.
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This article is the second of a series of articles detailing the development of near-infrared (NIR) methods for solid dosage-form analysis. Experiments were conducted at the Duquesne University Center for Pharmaceutical Technology to demonstrate a method for developing and validating NIR models for the analysis of active pharmaceutical ingredient (API) content and hardness of a solid dosage form. Robustness and cross-validation testing were used to optimize the API content and hardness models.
When you write up your observations and notes in a traditional laboratory notebook, there are a number of features that help to ensure data integrity. The notebook pages are numbered sequentially and they are bound together. Therefore, if a page is removed, it is obvious immediately. When recording observations in the notebook, the order of the write-up is important and is enforced by the sequential or linear pagination of the notebook. The author signs and dates the recorded information and this is witnessed by a second person, who signs and also dates when they reviewed the work. Note that it is not usual to time and date signatures in a laboratory notebook.
Working in an Electronic Environment
Many industry professionals know that analytical testing for biopharmaceuticals for all raw materials, production in-process stages, and final containers must be validated, and they generally understand how this can be achieved. Many of us even understand the basic concepts of laboratory compliance and production process quality. However, how exactly are analytical test method performance and process robustness related and how do they depend on each other? Furthermore, how do we monitor and maintain the accuracy and reliability of analytical methods long after validation completion to ensure the suitability of these methods for measuring process quality?
During pharmaceutical development and postapproval, it is often necessary to change analytical methods to ensure they remain stability-indicating, take advantage of improved analytical technology, monitor new related substances as a result of changes in synthetic or formulation processes, and improve analytical effeciency (e.g.,through automation).
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This article describes a software solution for automating the chromatographic method validation process starting from experimental planning, data acquisition and processing, through final report generation in a seamless manner. All experimental planning and calculations are accomplished within the chromatography data software and, thus, are structurally validated, secure, and audit trailed. Highlights of the software are provided, including benefits to the analyst. The analysis of important method validation characteristics such as linearity, accuracy, and precision is automated. These characteristics and their acceptance criteria can be captured in a method template, which adheres to the company's standard operating procedure. This template method can then be used repeatedly by other scientists in the organization, hence, eliminating the need to create a new experimental plan each time a new validation is conducted
This article provides guidance for minimally acceptable method validation practices and a foundation for assessing the risks and benefits associated with method validation programs.
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The chromatographic analytical method validation process involves a series of activities that are currently conducted in separate “technology islands” using tools that exist for each activity.This article describes a software program that provides an overarching automation technology for analytical method validation and brings together the individual activities under one integrated-technology platform that is adapted to multiple instruments and data systems.
Dissolution testing of poorly soluble compounds in immediate- release
(IR) solid dosage forms poses many challenges. These challenges include
developing and validating the test method, ensuring that the method is
appropriately discriminatory, and addressing the potential for an in
vivo–in vitro relationship (IVIVR) or correlation (IVIVC). The
objectives of dissolution testing, in general, vary during the life
cycle of a dosage form. The primary objective during Phases
0 and I is to develop a method to clearly establish the mechanism of in
vitro drug release and solubilization.During Phases II and III, the
objective shifts to identifying a test method that can provide an
IVIVR, IVIVC, or other biorelevant information. At registration and
goal is to identify a quality control (QC) dissolution test method to verify process and product consistency.
To ensure compliance with quality and safety standards, the United States, Europe,Japan, and other countries have published compendia, or pharmacopeias, that describe official test methods for many marketed drug products. For example, compendial analytical methods found in United States Pharmacopeia 25 (USP 25) are legally recognized analytical procedures under section 501 (b) of the Federal Food, Drug, and Cosmetic Act. For these compendial methods, USP provides regulatory guidance for method validation (1). In addition, validation of analytical methods is covered by the United States Code of Federal egulations (CFR). Specific references are 21 CFR 211.165 (e) and 21 CFR 211.194 (a).
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Changes were made to the USP method for the preparation of assay samples of ibuprofen (IBP) tablets (1). These changes included extraction and filtration techniques during sample preparation and required validation for the quantitation of IBP in assay samples. In this article, which is the first in a series about the validation of changes to the USP IBP assay method, we describe the validation of sample extraction and filtration techniques. Specifically, the extraction and filtration validation addresses the following:
● the effect of direct extraction (versus the powdering of tablets
and extraction or the shaking of coated tablets with glass beads
as per USP method) and shaking time on the disintegration
of tablets in extraction solvent and the solubilization of the
active ingredient as determined by the recovery of IBP from
● the effect of filtration (versus a centrifugation technique as
The sophistication of laboratory analysis instrumentation has advanced steadily over the past few decades.Much to the dismay of the manufacturing industry, this growth has tended to stay in the research arena. Recently, though, instrumentation companies have made strides to bring analytical technology from the laboratory into the pharmaceutical processing line. From the receipt of raw materials to in-process quality control testing, companies are developing unique analytical tools that are intended for easy integration into the pharmaceutical production environment.
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