This is a working overview of stationary phase, written for readers who want more than a one-paragraph summary but less than a textbook.
This page was last updated on 2026-03-16 and is reviewed periodically as new material appears.
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.
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.
| Property | Value | Notes |
|---|---|---|
| Column particle size | 3–5 µm for conventional HPLC; sub-2 µm for UHPLC | Smaller particles increase backpressure and efficiency. |
| Typical flow rate | 0.5–2.0 mL/min for a 4.6 mm internal diameter column | Flow scales with column diameter and particle size. |
| UV detection wavelength | 190–400 nm | Selection depends on analyte chromophore. |
| Column temperature | 25–40 °C | Temperature affects retention, selectivity, and pressure. |
| Injection volume | 1–20 µL | Larger volumes may distort early-eluting peaks. |
Method validation demonstrates that an HPLC procedure is suitable for its intended purpose. Common validation parameters include accuracy, precision, specificity, linearity, range, limit of detection, limit of quantification, and robustness. Accuracy reflects agreement with a reference value, while precision describes repeatability under defined conditions. Specificity shows whether the method can measure the analyte in the presence of impurities or matrix components. Validation documents are reviewed before a method is used for routine testing or regulatory submissions.
System suitability testing is performed before and during analytical runs to confirm that the instrument and method are working as expected. Typical checks include retention time, peak area precision, resolution between critical pairs, tailing factor, and theoretical plate count. Acceptance criteria are set in the method or pharmacopeial monograph. If a suitability check fails, the run may be rejected and the instrument or sample preparation may need investigation. This practice helps prevent release of data from a system that has drifted out of control.
Quality control samples are inserted at intervals to monitor accuracy and precision throughout a batch. Blank samples detect contamination, while spiked samples assess recovery from the sample matrix. Calibration standards establish the relationship between detector response and concentration, and control samples are prepared independently from them whenever possible. Laboratories also participate in proficiency testing and maintain audit trails, instrument logs, and reagent records. Ongoing review of control charts can reveal trends before they cause out-of-specification results.
Method validation establishes that an HPLC procedure is suitable for its intended use. Key parameters include accuracy, precision, specificity, linearity, range, limit of detection, limit of quantitation, and robustness. Accuracy measures agreement with a true or accepted value, while precision describes repeatability and intermediate precision. Specificity confirms that the method measures the analyte without interference from impurities, degradants, or excipients. Validation is documented in a protocol and report, and acceptance criteria are set before experiments begin. Regulatory guidance varies by region, but the general principles are widely harmonized.
System suitability testing is performed before and during analytical runs to confirm that the instrument and method are working as expected. Common checks include retention time, peak area, resolution between critical pairs, tailing factor, and theoretical plate count. Results are compared with predefined limits, and a failed check requires investigation before sample results are reported. Quality control samples at low, middle, and high concentrations are injected at intervals to monitor accuracy and precision. Blank injections detect carryover and contamination, while control charts track performance over time.
Number average molar mass (Mn), also loosely referred to as number average molecular weight (NAMW). Mass average molar mass (Mw), where w stands for weight; also commonly referred to as weight average or weight average molecular weight (WAMW). Z-average molar mass (Mz), where z stands for centrifugation (from German Zentrifuge). Viscosity average molar mass (Mv).
== External links == LPAR6 human gene location in the UCSC Genome Browser. LPAR6 human gene details in the UCSC Genome Browser. This article incorporates text from the United States National Library of Medicine, which is in the public domain.
== Parent isotope source == 99Mo can be obtained by the neutron activation (n,γ reaction) of 98Mo in a high-neutron-flux reactor. However, the most frequently used method is through fission of uranium-235 in a nuclear reactor. While most reactors currently engaged in 99Mo production use highly enriched uranium-235 targets, proliferation concerns have prompted some producers to transition to low-enriched uranium targets. The target is irradiated with neutrons to form 99Mo as a fission product (with 6.1% yield). Molybdenum-99 is then separated from unreacted uranium and other fission products in a hot cell.
Sources: en.wikipedia.org
In 2015 fire and heat resulted in 67 million injuries. This resulted in about 2.9 million hospitalizations and 238,000 dying. This is down from 300,000 deaths in 1990. This makes it the fourth leading cause of injuries after motor vehicle collisions, falls, and violence. About 90% of burns occur in the developing world. This has been attributed partly to overcrowding and an unsafe cooking situation. Overall, nearly 60% of fatal burns occur in Southeast Asia with a rate of 11.6 per 100,000. The number of fatal burns has decreased from 280,000 in 1990 to 176,000 in 2015. In the developed world, adult males have twice the mortality as females from burns. This is most probably due to their higher risk occupations and greater risk-taking activities. In many countries in the developing world, however, females have twice the risk of males. This is often related to accidents in the kitchen or domestic violence. In children, deaths from burns occur at more than ten times the rate in the developing than the developed world. Overall, in children it is one of the top fifteen leading causes of death. From the 1980s to 2004, many countries have seen both a decrease in the rates of fatal burns and in burns generally.
== Early life == Rosanna Jeanne Reardon was born on June 8, 1985, in Seattle, Washington, where she was raised alongside her younger sister Molly Lu. She is of Italian, Croatian, German, and Irish ancestry. She was baptized Catholic. Throughout her childhood, she was known by friends and family as a "nerdy", "awkward" kid, and she struggled with dyslexia and ADHD in school. She originally wanted to become an actress, and after attending West Seattle High School and Pacific Lutheran University, she moved to Los Angeles to pursue acting. Prior to her YouTube career, Pansino worked as a teacher in China and as a Methodist church secretary. Pansino's interest in cooking was inspired by her father and grandmother, and she stated in 2014 that "...baking has always been a hobby of mine. I have always loved being creative with my food and making themed sweets for my friends."
==== Image analysis ==== Both art-historical digital image processing and analog techniques have been applied to the shroud images. In 1976 scientists used imaging equipment from the American National Aeronautics and Space Administration (NASA) to analyze a photograph of the Shroud image and decoded the shroud image into a three-dimensional image. The optical physicist and former STURP member John Dee German has noted that it is not difficult to make a photograph which has 3D qualities. If the object being photographed is lit from the front, and a non-reflective "fog" of some sort exists between the camera and the object, then less light will reach and reflect back from the portions of the object that are farther from the lens, thus creating a contrast which is dependent on distance. The front image on the shroud is 1.95 metres (6 ft 5 in) long, and is not exactly the same size as the rear image, which is 2.02 metres (6 ft 8 in) long. Analysis of the images found them to be compatible with the shroud having been used to wrap a body 1.75 metres (5 ft 9 in) long. The image could be compared to oshiguma, the making of face-prints as an artform, in Japan. Furthermore, the subject's physical appearance corresponds to Byzantine iconography. The Shroud cloth is composed of threads of a nominal diameter of 0.15 mm, woven with fibers of linen with a diameter of about 10-20 μm. The Shroud image is a faint and superficial image caused by a translucent and discontinuous yellow discoloration of the fibers.
Sources: en.wikipedia.org
Quantitative studies of enzymatic reactions often found that the acceleration of chemical reaction speed cannot be fully explained by existing theories like the approximation, acid/base catalysis and electrophile/nucleophile catalysis. And there is an obvious paradox: in reversible enzymatic reaction if the active site perfectly fits the substrates then the backward reaction will be slowed since products cannot fit perfectly into the active site. So conformational distortion was introduced and argues that both active site and substrate can undergo conformational changes to fit with each other all the time.
The neptunium antimonide compound NpSb was produced in 1971 by placing equal quantities of both elements in a vacuum tube, heating them to the melting point of antimony, and then heating it further to 1000 °C for sixteen days. This procedure also produced trace amounts of an additional antimonide compound Np3Sb4. One neptunium-bismuth compound, NpBi, has also been reported. The neptunium carbides NpC, Np2C3, and NpC2 (tentative) have been reported, but have not characterized in detail despite the high importance and utility of actinide carbides as advanced nuclear reactor fuel. NpC is a non-stoichiometric compound, and could be better labelled as NpCx (0.82 ≤ x ≤ 0.96). It may be obtained from the reaction of neptunium hydride with graphite at 1400 °C or by heating the constituent elements together in an electric arc furnace using a tungsten electrode. It reacts with excess carbon to form pure Np2C3. NpC2 is formed from heating NpO2 in a graphite crucible at 2660–2800 °C.
Parental concern about vaccines has led to a decreasing uptake of childhood immunizations and an increasing likelihood of measles outbreaks. The refrigerator mother theory has been conclusively refuted by scientific evidence.
Sources: en.wikipedia.org
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.
UHPLC uses columns with smaller particles and operates at higher pressures than conventional HPLC. These conditions can improve speed, resolution, and sensitivity. Both techniques use the same fundamental separation principles.
Validation shows that a method performs reliably for its intended purpose across a defined range. It assesses accuracy, precision, specificity, linearity, and robustness. Regulated testing often requires documented validation before routine use.
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.