A practical reference on method validation: what it is, how it behaves, what the literature reports, and where the honest uncertainties sit.
Reviewed 2025-11-18. Anything still debated is marked as such rather than presented as settled.
Developing an HPLC method begins with defining the purpose, such as quantifying a main component, measuring impurities, or confirming identity. Analysts select separation mode, column, mobile phase, detection, and sample preparation based on analyte properties and matrix. Experiments vary solvent strength, pH, buffer type, and temperature to achieve resolution between critical peaks. The goal is a robust method that produces reliable results across instruments and operators. Method development often involves trial runs and statistical optimization.
Validation demonstrates that a method is suitable for its intended use. Typical performance characteristics include accuracy, precision, specificity, linearity, range, limit of detection, limit of quantitation, and robustness. Regulators and standards organizations provide frameworks, but specific requirements depend on the application and jurisdiction. System suitability tests are run before sample analysis to confirm resolution, peak symmetry, retention time repeatability, and sensitivity. A validated method is not permanently fixed; changes may require partial or full revalidation.
Separation in HPLC depends on the chemistry of the stationary phase, the composition of the mobile phase, and the physical properties of the column. Reverse-phase separations use a nonpolar stationary phase and a polar mobile phase, and they are common for many organic compounds. Ion-exchange, size-exclusion, and normal-phase modes serve other classes of analytes. Gradient elution changes solvent strength over time, while isocratic elution holds it constant. Flow rate, temperature, particle size, and column length all influence peak shape and resolution. Detection may use ultraviolet absorbance, fluorescence, refractive index, or mass spectrometry, depending on the analyte and the required sensitivity.
Routine HPLC testing compares a sample result with a calibration curve prepared from known reference standards. Peak area or peak height is plotted against concentration, and the curve is used to estimate unknown amounts. Retention time supports tentative identification when compared with a standard, though mass spectrometry or another confirmatory method may be needed for definitive identification. Pre-run checks verify repeatability, resolution, and peak symmetry before sample analysis. Limits of detection and quantification describe the smallest amounts that can be reliably observed or measured. Sample preparation, filtration, and degassing help prevent column damage and inconsistent results.
| Property | Value | Notes |
|---|---|---|
| Validation parameter | Accuracy | Closeness of measured value to accepted reference value |
| Validation parameter | Precision | Agreement among repeated measurements under specified conditions |
| System suitability check | Resolution ≥ 1.5 | Baseline separation between critical peak pair |
| System suitability check | Tailing factor ≤ 2.0 | Common target for peak symmetry |
| Documentation | Validation report | Summarizes experiments, acceptance criteria, and conclusions |
Quality control laboratories use HPLC to check identity, purity, concentration, and stability of raw materials and finished products. A validated method specifies the column, mobile phase, flow rate, detection wavelength, injection volume, and run time. Samples are prepared and compared against reference standards of known concentration. The resulting chromatogram provides quantitative data, such as assay values and impurity levels. This approach is common in pharmaceutical, food, environmental, and industrial testing where consistent measurements are required.
Method validation demonstrates that an analytical procedure is suitable for its intended purpose. Typical validation characteristics include accuracy, precision, specificity, linearity, range, limit of detection, limit of quantitation, and robustness. Regulatory guidance from bodies such as the International Council for Harmonisation and the United States Pharmacopeia outlines expectations, though specific criteria depend on the product and method. System suitability tests are run before sample analysis to confirm resolution, peak symmetry, column efficiency, and injection repeatability. Failure of these checks can invalidate a batch of measurements.
Practical HPLC testing depends on careful sample preparation and instrument maintenance. Samples may require filtration, dilution, pH adjustment, or extraction to avoid column damage and matrix interference. Mobile phases are degassed and filtered, and columns are equilibrated before injection. Common problems include peak tailing, baseline drift, ghost peaks, carryover, and co-elution of analytes. Documentation of instrument logs, calibration records, and electronic audit trails supports data integrity and traceability. Ongoing training and routine maintenance help reduce variability between analysts and laboratories.
Quality control for HPLC testing combines scheduled checks, documented procedures, and review of results. Before sample analysis, system suitability testing confirms that the instrument, column, and method meet predefined criteria. Common criteria include resolution between critical peaks, retention time precision, peak tailing, and theoretical plate count. Failure triggers investigation before results are reported. Records link raw data, calculations, instrument logs, and analyst identity to each batch, supporting audits and repeat analysis.
Method validation evaluates accuracy, precision, specificity, linearity, range, detection limit, quantitation limit, and robustness. Regulatory guidance for pharmaceuticals, foods, and environmental testing defines expected documentation and acceptance criteria. Verification confirms that a validated method works in a specific laboratory with its own instruments and reagents. Calibration curves use reference standards with known purity and traceability, while measurement uncertainty is estimated from validation data, control charts, and collaborative studies. The scope of validation depends on the method's intended use.
Routine quality control monitors retention time shifts, baseline noise, system pressure, and peak shape. Trends can reveal column aging, mobile phase preparation errors, detector drift, or sample degradation. Corrective actions may include replacing the column, preparing fresh mobile phase, or recalibrating the detector. Stability testing often uses HPLC to measure parent compound loss and degradation product formation. Open questions remain about how accelerated stability results extrapolate to long-term storage under varied conditions.
Kief is a powder, rich in trichomes, that can be sifted from the leaves, flowers and fruits of cannabis plants and either consumed in powder form or compressed to produce cakes of hashish. The term derives from colloquial Arabic كيف kēf/kīf, meaning pleasure.
== Editorship of journals == Editor-in-Chief, Journal of Current Biomedical Research (2017 to date) Associate Editor of African Journal of Pharmaceutical Sciences and Pharmacy Associate Editor, Journal of Pharmaceutical and Allied Sciences Member Editorial Board of American Journal of Pharmacotherapy and Pharmaceutical Sciences (2024)
==== Liquid crystal microcapsules ==== Fabrication of liquid crystals has been a point of intrigue for over 5 decades for their anisotropic properties. Microfluidic devices can be used for the synthesis of confined cholesteric liquid crystals through layering of multiple shell-like layers consisting of varying oil-in-water and water-in-oil emulsions. The construction of encapsulated liquid crystals through microfluidic flow of liquid crystal droplets in an immiscible oil allows for monodispersed emulsion layer thickness and composition previously unseen before the advent of microfluidic techniques. The advent of liquid crystal technology has led to advancements in optical displays used both in research and consumer branded products, but more recent discoveries have opened the door to the combination of photon combination and upconversion as well as optical sensors for biological analytes.
Providing a sense of scale, a forest 62,000 square kilometres (24,000 sq mi) in area, the size of the U.S. state of Pennsylvania, is estimated to emit 3.4 million kg (7.5 million lb) of terpenes on a typical August day during the growing season. Maize produces the VOC (Z)-3-hexen-1-ol and other plant hormones. The taste of bitterness, found in foods such as olives, coffee and dark chocolate is caused by detection of VOCs by taste receptors.
Sources: en.wikipedia.org
=== Spain === All Spanish medical degree holders need to pass a competitive national exam (named 'MIR') in order to access the specialty training program. This exam gives them the opportunity to choose both the specialty and the hospital where they will train, among the hospitals in the Spanish Healthcare Hospital Network. Currently, medical specialties last from 4 to 5 years. There are plans to change the training program system to one similar to the UK's. There have been some talks between Ministry of Health, the Medical College of Physicians and the Medical Student Association but it is not clear how this change process is going to be.
In the United States, 49 states and the District of Columbia have expanded naloxone access at a pharmacy level via standing order, protocol order, naloxone-specific collaborative practice agreement, or pharmacist prescriptive authority.
Irradiation of food is the exposure of food to ionizing radiation. Multiple types of ionizing radiation can be used, including beta particles (high-energy electrons) and gamma rays (emitted from radioactive sources such as cobalt-60 or cesium-137). Irradiation can kill bacteria, molds, and insect pests, reduce the ripening and spoiling of fruits, and at higher doses induce sterility. The technology may be compared to pasteurization; it is sometimes called "cold pasteurization", as the product is not heated. Irradiation may allow lower-quality or contaminated foods to be rendered marketable. National and international expert bodies have declared food irradiation as "wholesome"; organizations of the United Nations, such as the World Health Organization and Food and Agriculture Organization, endorse food irradiation. Consumers may have a negative view of irradiated food based on the misconception that such food is radioactive; in fact, irradiated food does not and cannot become radioactive. Activists have also opposed food irradiation for other reasons, for example, arguing that irradiation can be used to sterilize contaminated food without resolving the underlying cause of the contamination. International legislation on whether food may be irradiated or not varies worldwide from no regulation to a full ban. Approximately 500,000 tons of food items are irradiated per year worldwide in over 40 countries. These are mainly spices and condiments, with an increasing segment of fresh fruit irradiated for fruit fly quarantine.
Sources: en.wikipedia.org
== New roles == Recent findings suggest that StAR may also traffic cholesterol to a second mitochondrial enzyme, sterol 27-hydroxylase. This enzyme converts cholesterol to 27-hydroxycholesterol. In this way it may be important for the first step in one of the two pathways for the production of bile acids by the liver (the alternative pathway). Evidence also shows that the presence of StAR in a type of immune cell, the macrophage, where it can stimulate the production of 27-hydroxycholesterol. In this case, 27-hydroxycholesterol may by itself be helpful against the production of inflammatory factors associated with cardiovascular disease. No study has yet found a link between the loss of StAR and problems in bile acid production or increased risk for cardiovascular disease. Recently StAR was found to be expressed in cardiac fibroblasts in response to ischemic injury due to myocardial infarction. In these cells it has no apparent de novo steroidogenic activity, as evidenced by the lack of the key steroidogenic enzymes cytochrome P450 side chain cleavage (CYP11A1) and 3 beta hydroxysteroid dehydrogenase (3βHSD). StAR was found to have an anti-apoptotic effect on the fibroblasts, which may allow them to survive the initial stress of the infarct, differentiate and function in tissue repair at the infarction site.
Osedax antarcticus Glover, Wiklund & Dahlgren, 2013 Osedax bozoi Berman, Hiley, Read & Rouse, 2024 Osedax braziliensis Fujiwara, Jimi, Sumida, Kawato, Kitazato Osedax bryani Rouse, Goffredi, Johnson & Vrijenhoek Osedax byronbayensis Georgieva, Wiklund, Ramos, Neal, Glasby & Gunton, 2023 Osedax craigmcclaini Berman, Hiley, Read, Rouse, 2024 Osedax crouchi Amon, Wiklund, Dahlgren, Copley, Smith, Jamieson & Glover, 2014 Osedax deceptionensis Taboada, Cristobo, Avila, Wiklund & Glover, 2013 Osedax docricketts Rouse, Goffredi, Johnson & Vrijenhoek Osedax estcourti Berman, Hiley, Read & Rouse, 2024 Osedax fenrisi Eilertsen, Dahlgren & Rapp, 2020 Osedax frankpressi Rouse, Goffredi & Vrijenhoek, 2004 Osedax jabba Rouse, Goffredi, Johnson & Vrijenhoek Osedax japonicus Fujikura, Fujiwara & Kawato, 2006 Osedax knutei Rouse, Goffredi, Johnson & Vrijenhoek Osedax lehmani Rouse, Goffredi, Johnson & Vrijenhoek Osedax lonnyi Rouse, Goffredi, Johnson & Vrijenhoek Osedax mucofloris Glover, Kallstrom, Smith & Dahlgren, 2005 Osedax nataliae Gularte, Sumida, Bergamo & Rouse, 2024 Osedax nordenskjoeldi Amon, Wiklund, Dahlgren, Copley, Smith, Jamieson & Glover, 2014 Osedax priapus Rouse et al., 2014 Osedax packardorum Rouse, Goffredi, Johnson & Vrijenhoek Osedax randyi Rouse, Goffredi, Johnson & Vrijenhoek Osedax rogersi Amon, Wiklund, Dahlgren, Copley, Smith, Jamieson & Glover, 2014 Osedax roseus Rouse, Worsaae, Johnson, Jones & Vrijenhoek, 2008 Osedax rubiplumus Rouse, Goffredi & Vrijenhoek, 2004 Osedax ryderi Rouse, Goffredi, Johnson & Vrijenhoek Osedax sigridae Rouse, Goffredi, Johnson & Vrijenhoek Osedax talkovici Rouse, Goffredi, Johnson & Vrijenhoek Osedax tiburon Rouse, Goffredi, Johnson & Vrijenhoek Osedax traceyae Berman, Hiley, Read & Rouse, 2024 Osedax ventana Rouse, Goffredi, Johnson & Vrijenhoek Osedax waadjum Georgieva, Wiklund, Ramos, Neal, Glasby & Gunton, 2023 Osedax westernflyer Rouse, Goffredi, Johnson & Vrijenhoek
Pizza Hut also offers a chicken wing menu. The menu was launched in 2003 under the WingStreet branding in existing Pizza Hut franchises, with the plan to turn WingStreet into a standalone chain. Yum! Brands predicted aggressive growth, adding more than 4,000 locations by 2010. In 2012, the company opened a standalone pilot store in Denton, Texas. The store was unsuccessful and closed the following year.
Sources: en.wikipedia.org
It is a set of checks performed before or during an HPLC run to confirm the system works as expected. Parameters may include resolution, tailing factor, theoretical plates, and retention time precision. Failure can trigger maintenance, method adjustment, or repeat analysis.
Validation follows a planned protocol that tests accuracy, precision, specificity, linearity, range, detection limits, quantitation limits, and robustness. Results are compared against predefined acceptance criteria. The validation report supports regulatory filing or routine use.
Revalidation may be needed after changes to column chemistry, mobile phase, detection, sample preparation, or instrument type. It can also follow a pattern of out-of-specification results. The scope depends on whether the change affects method performance.
HPLC testing measures the presence and amount of one or more compounds in a liquid sample. It separates mixture components and records detector responses as peaks, which are compared with reference standards. Results are usually reported as concentrations or relative percentages.