If you have been reading about Stationary phase and want a single page that covers the useful parts, this is it: definitions, context, how it is studied, and the questions that come up repeatedly.
Last reviewed on 2026-04-24. Where a claim depends on a specific study, the study is described rather than over-claimed.
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 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.
Several separation modes exist, including reversed-phase, normal-phase, ion-exchange, size-exclusion, and hydrophilic interaction liquid chromatography. Reversed-phase uses a nonpolar stationary phase with a polar mobile phase and is widely applied to small organic molecules. Gradient elution changes mobile phase composition during the run, while isocratic elution keeps it constant. Column chemistry, particle size, temperature, flow rate, and mobile phase pH all influence retention and resolution. Method development selects conditions that separate analytes from matrix components and from each other.
Detection commonly uses ultraviolet-visible absorbance, fluorescence, refractive index, or mass spectrometry. Ultraviolet detection depends on molecular chromophores that absorb light at specific wavelengths. Mass spectrometry provides mass information and sensitive quantification, often after electrospray ionization. Before sample batches, performance checks examine resolution, elution time repeatability, peak symmetry, and plate count. Matrix effects and co-elution remain recognized uncertainties; formal validation studies and orthogonal detection help address them. Detector choice depends on analyte properties and required sensitivity.
| Parameter | Typical acceptance criterion | Notes |
|---|---|---|
| Resolution | ≥ 1.5 | Baseline separation of adjacent peaks |
| Tailing factor | ≤ 2.0 | Peak symmetry measure |
| Theoretical plates | > 2000 | Column efficiency indicator |
| Injection repeatability | ≤ 2% RSD | Relative standard deviation for replicate injections |
| Linearity | r² ≥ 0.995 | Calibration curve over the working range |
HPLC testing is an analytical technique used to separate, identify, and quantify components in a liquid sample. It relies on a pressurized mobile phase that carries the sample through a column packed with stationary phase. Different compounds travel at different rates because of interactions with the stationary and mobile phases. The resulting signal versus time is a chromatogram. Peak position indicates identity under specified conditions, while peak area or height relates to amount.
Laboratories apply HPLC testing across pharmaceutical, food, environmental, and industrial chemistry. The method can measure active ingredients, impurities, additives, preservatives, and degradation products. Sample preparation often includes dilution, filtration, and sometimes extraction or derivatization. The choice of column, mobile phase, pH, temperature, and detector depends on the analytes and matrix. Results are compared with reference standards to assign identity and concentration. Method suitability is judged by resolution, precision, and accuracy.
High-performance liquid chromatography is an analytical technique that separates components in a liquid sample. A pump moves a liquid mobile phase through a column packed with a solid stationary phase. Compounds interact differently with both phases and travel at different rates, leaving the column at distinct retention times. A detector records these arrivals as peaks on a chromatogram. The resulting pattern supports identification and quantification of substances in mixtures. Modern instruments use high pressure to force solvent through small particles, which improves speed and resolution compared with older low-pressure liquid chromatography methods.
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.
ACE in plasma was discovered by Leonard T. Skeggs, Norman Shumway, and their colleagues in 1956, but at that time there was no known physiological basis for its use in heart failure treatment. That changed in 1962, when the physiological rationale for using vasodilators to reduce afterload in heart failure was established by American cardiologist Edmund Sonnenblick through force-velocity studies of isolated cardiac muscle. These studies demonstrated that the resistance against which the ventricle contracts directly determines its performance—providing the theoretical justification for afterload reduction as a therapeutic strategy that ACE inhibitors would later exploit. Around this time, it was also noted that those who worked in banana plantations in South-western Brazil collapsed after being bitten by a pit viper, leading to a search for a blood pressure lowering component in its venom. Brazilian scientist Sérgio Henrique Ferreira reported a bradykinin-potentiating factor (BPF) present in the venom of Bothrops jararaca, a South American pit viper, in 1965. Ferreira then went to John Vane's laboratory as a postdoctoral fellow with his already-isolated BPF. The conversion of the inactive angiotensin I to the potent angiotensin II was thought to take place in the plasma. However, in 1967, Kevin K. F. Ng and John R. Vane showed plasma ACE is too slow to account for the conversion of angiotensin I to angiotensin II in vivo. Subsequent investigation showed rapid conversion occurs during its passage through the pulmonary circulation.
== Culture == Most of the district's news media has been either weekly or monthly publications. The earliest newspaper was the Scarborough News and Advertiser which was published weekly starting in September 1921, which lasted until the 1930s. Other short-lived papers and magazines included The Enterprise (1945–1966), Scarborough Mail (1946–1955) and The News (1952–1996) and 54east magazine (2005–2009). The last remaining English-language local newspaper was the Scarborough Mirror, which started publication in 1962 and was later acquired by the Toronto Star's community news division, Metroland. In 2023, it became an online-only publication as part of Toronto.com. A Scarborough edition of the Toronto-wide photography publication SNAP Scarborough was launched in 2009. Ming Pao Daily News is a Chinese-language newspaper whose headquarters is in the district. They started publication in 1993. In 1961, the CTV Toronto (CFTO) television channel began operating from the 9 Channel Nine Court studios at the intersection of McCowan Road and Highway 401. Today, the studio also is the headquarters of The Sports Network (TSN). Both companies produce programming at the studio. In 1970, Trillium Cable started to provide cable TV service to Scarborough. It was purchased by Shaw Cable in 1995; but later swapped with Rogers Cable in 2001. During the early days of the company, they produced several local shows for their own cable channel. These shows were produced by volunteers and showed a wide variation in quality. These shows were satirized by Mike Myers in his comedy film Wayne's World.
February 6: Law modifying the rules for legal separation. July 15: Law on free medical assistance for people without means: women giving birth are considered equivalent to sick persons. August 19: A law concerning fire safety measures in certain regions of France holds the husband civilly liable for offenses and violations committed by his wife. July 20, 1895: Law on savings banks. December 7, 1897: Law granting women the right to be witnesses in civil status acts and in official documents in general. 1898
On 20 March, Lula and health minister, Nísia Trindade, announced the re-creation of the Mais Médicos programme, first created in 2013 with the purpose of expanding the number of health professionals in less economically developed areas and in the interior of the country. The programme had been partially replaced by Bolsonaro's "Doctors for Brazil"; the "Mais Médicos para o Brasil", as it is now called, should prioritize Brazilian professionals and, according to Paulo Pimenta, chief minister of the Secretary of Social Communication, increase the number of health professionals and improve the Sistema Único de Saúde (SUS).
Sources: en.wikipedia.org
== History of the term == The term "RuBisCO" was coined humorously in 1979, by David Eisenberg at a seminar honouring the retirement of the early, prominent RuBisCO researcher, Sam Wildman, and also alluded to the snack food trade name "Nabisco" in reference to Wildman's attempts to create an edible protein supplement from tobacco leaves. The capitalization of the name has been long debated. It can be capitalized for each letter of the full name (Ribulose-1,5 bisphosphate carboxylase/oxygenase), but it has also been argued that is should all be in lower case (rubisco), similar to other terms like scuba or laser.
According to Ukraine, 55 Russian missiles were fired at targets in Ukraine, along with another 24 Shahed-136 drones. The Ukrainian Air Force claimed to have shot down all of the drones and 47 of the missiles. Among the weapons used in the attack was a Kh-47 Kinzhal hypersonic missile. Kyiv's mayor said one person died and two were wounded when an apartment block was hit in the Holosiivskyi District. Across the country, 11 people were killed and 11 more injured, according to the Emergency Service. This was the 13th mass attack since the invasion began.
Blue tangs experience three different social modes: territorial, schooling, and wandering. Blue tangs in non-territorial modes form schools and wander, while territorial blue tangs do not. Territoriality reduces competition for food resources because one individual claims both a territory and its resources. Additionally, schooling allows fish to better overcome food defense by others, and wandering allows for individual movement to feeding areas, cleaning stations, and other resources. Social behaviors are affected by outside conditions such as damselfish density, conspecific population density, and life history stage. Those in the territorial mode are aggressive and actively chase intruding blue tangs. They swim slower and feed more frequently than non-territorial blue tangs. Schooling fish form compact groups with other blue tangs in addition to other species. Territorial blue tangs are found most often in the reef flat zone (sandy-bottomed with rocks or coral) and are rarely found in the spur zone (low coral ridges) or groove zone (sandy bottom channels). Territoriality prevalence decreases with increasing damselfish population, and the time of day also does not affect territoriality prevalence. Fish in schooling mode swim fast, eat at intermediate rates, and are not aggressive, even though they are often chased by damselfish. Schooling is most prevalent in areas with high damselfish densities such as the groove zone, spur zone, and crest zone (shallow reef area), while schools are most prevalent near midday. Blue tangs are often seen schooling with midnight parrotfish.
Sources: en.wikipedia.org
A Komodo dragon may attempt to speed up the process by ramming the carcass against a tree to force it down its throat, sometimes ramming so forcefully that the tree is knocked down. A small tube under the tongue that connects to the lungs allows it to breathe while swallowing. After eating up to 80% of its body mass in one meal, it drags itself to a sunny location to speed digestion, as the food could rot and poison the dragon if left undigested in its stomach for too long. Because of their slow metabolism, large dragons can survive on as few as 12 meals a year. After digestion, the Komodo dragon regurgitates a mass of horns, hair, and teeth known as the gastric pellet, which is covered in malodorous mucus. After regurgitating the gastric pellet, it rubs its face in the dirt or on bushes to get rid of the mucus.
=== New generation fuel === The aim of Guy Reinaud's project is to reply to the needs, in renewable domestic energy, of a population of 2 billion individuals living in the tropical regions of Africa, Latin America and Asia, and at the same time fighting against deforestation and climatic changes. The proposed solution is biomass charcoal as a source of renewable energy - patented and developed by Pro-Natura International. Biomass charcoal produces domestic energy from non-upgraded biomass, other than wood, in particular agricultural residues non-consumed by animals. By reducing the dependence on wood, biomass charcoal protects forests and as a consequence fights against desertification. Moreover, the energy produced by biomass charcoal is equal to that of wood charcoal for a sales price per kilo 50% cheaper.
With the widespread interest in birds, use of a large number of people to work on collaborative ornithological projects that cover large geographic scales has been possible. These citizen science projects include nationwide projects such as the Christmas Bird Count, Backyard Bird Count, the North American Breeding Bird Survey, the Canadian EPOQ or regional projects such as the Asian Waterfowl Census and Spring Alive in Europe. These projects help to identify distributions of birds, their population densities and changes over time, arrival and departure dates of migration, breeding seasonality, and even population genetics. The results of many of these projects are published as bird atlases. Studies of migration using bird ringing or colour marking often involve the cooperation of people and organizations in different countries.
Sources: en.wikipedia.org
Method validation is the documented process of confirming that an HPLC procedure is suitable for its intended use. It evaluates accuracy, precision, specificity, linearity, range, detection limits, and robustness. Validation criteria depend on the regulatory context and the sample type.
System suitability tests are short checks performed before or during an HPLC run to verify instrument and method performance. They often include resolution, tailing factor, theoretical plates, and injection precision. Results must meet predefined limits for sample data to be accepted.
HPLC retention time alone cannot definitively identify an unknown substance. A match with a reference standard under identical conditions provides supporting evidence. Confirmation typically requires mass spectrometry, nuclear magnetic resonance, or another orthogonal technique.
HPLC separates and quantifies compounds in a liquid sample. Detectors produce a response proportional to the amount of a compound passing through the flow cell. Identification by retention time requires comparison with a known standard.