If you have been reading about HPLC 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 2026-03-15. Where a claim depends on a specific study, the study is described rather than over-claimed.
HPLC testing separates dissolved compounds by passing a liquid sample through a column packed with stationary phase. A pump delivers mobile phase at controlled flow, and the sample components interact differently with stationary and mobile phases. Compounds that spend more time in mobile phase elute earlier; those retained by stationary phase elute later. Detectors record elution as peaks, and peak area or height relates to amount. This mechanism underpins quantitative analysis of mixtures.
Most routine HPLC testing uses reversed-phase columns, where the stationary phase is nonpolar and the mobile phase is a polar mixture such as water with an organic solvent. Analytes partition between the two phases according to polarity, size, and charge. Gradients that change solvent composition over time can separate compounds with broad retention ranges. Isocratic conditions keep solvent composition constant and suit simpler mixtures. The choice of column chemistry, pH, and temperature affects selectivity and peak shape.
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.
| Property | Value | Notes |
|---|---|---|
| Separation mode | Reversed-phase | Nonpolar stationary phase with polar mobile phase |
| Typical column particle size | 3–5 µm | Smaller particles improve resolution but raise pressure |
| Typical flow rate | 0.5–2.0 mL/min | Depends on column dimensions and pressure limits |
| Common detection | UV-Vis absorbance | Requires analytes with chromophores |
| Typical run time | 5–30 min | Varies with method, gradient, and sample complexity |
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.
A characterization of these fibers was given, as well as hypotheses for their growth in a nitrogen atmosphere at low pressures. In 1981, a group of Soviet scientists published the results of chemical and structural characterization of carbon nanoparticles produced by a thermocatalytic disproportionation of carbon monoxide. Using TEM images and XRD patterns, the authors suggested that their "carbon multi-layer tubular crystals" were formed by rolling graphene layers into cylinders. They speculated that via this rolling, many different arrangements of graphene hexagonal nets are possible. They suggested two such possible arrangements: a circular arrangement (armchair nanotube); and a spiral, helical arrangement (chiral tube). In 1987, Howard G. Tennent of Hyperion Catalysis was issued a U.S. patent for the production of "cylindrical discrete carbon fibrils" with a "constant diameter between about 3.5 and about 70 nanometers..., length 102 times the diameter, and an outer region of multiple essentially continuous layers of ordered carbon atoms and a distinct inner core...." Helping to create the initial excitement associated with carbon nanotubes were Iijima's 1991 discovery of multi-walled carbon nanotubes in the insoluble material of arc-burned graphite rods; and Mintmire, Dunlap, and White's independent prediction that if single-walled carbon nanotubes could be made, they would exhibit remarkable conducting properties. Nanotube research accelerated greatly following the independent discoveries by Iijima and Ichihashi at NEC and Bethune et al.
=== 15 June === Three people were killed in a Russian cluster munitions attack in Ulakly, Donetsk Oblast. In Russia, the governor of Belgorod Oblast claimed that five people were killed by Ukrainian shelling in Shebekino. The US government announced $1.5 billion in humanitarian aid for Ukraine, mostly for its energy sector. The June 2024 Ukraine peace summit opened in Switzerland with 92 countries participating. Russia and China did not attend the meeting.
== See also == Anatomical pathology – Medical specialty Blood test – Laboratory analysis performed on a blood sample Clinical Laboratory Improvement Amendments – United States federal regulatory standards Diagnostics – Identification of the nature and cause of a certain phenomenonPages displaying short descriptions of redirect targets Healthcare – Prevention of disease and promotion of well-beingPages displaying short descriptions of redirect targets Histology – Study of the microscopic anatomy of cells and tissues of plants and animals Laboratory stewardship – Optimizing use of laboratory tests Pathology – Study of disease Point-of-care testing – Diagnostic testing performed at or near the point of care Associations American Society for Clinical Pathology – Medical association Association for Diagnostics & Laboratory Medicine – Global scientific society Journals Laboratory medicine journals
Sources: en.wikipedia.org
== Nuclear fuel cycle == The nuclear fission properties of berkelium are different from those of the neighboring actinides curium and californium, and they suggest berkelium to perform poorly as a fuel in a nuclear reactor. Specifically, berkelium-249 has a moderately large neutron capture cross section of 710 barns for thermal neutrons, 1200 barns resonance integral, but very low fission cross section for thermal neutrons. In a thermal reactor, much of it will therefore be converted to berkelium-250 which quickly decays to californium-250. In principle, berkelium-249 can sustain a nuclear chain reaction in a fast breeder reactor. Its critical mass is relatively high at 192 kg, which can be reduced with a water or steel reflector but would still exceed the world production of this isotope. Berkelium-247 can maintain a chain reaction both in a thermal-neutron and in a fast-neutron reactor, however, its production is rather complex and thus the availability is much lower than its critical mass, which is about 75.7 kg for a bare sphere, 41.2 kg with a water reflector and 35.2 kg with a steel reflector (30 cm thickness).
==== Betaferon (interferon beta-1b) ==== Closely related to interferon beta-1a is interferon beta-1b, which is also indicated for MS, but is formulated with a different dose and administered with a different frequency. Each drug has a different safety/efficacy profile. Interferon beta-1b is marketed only by Bayer in the US as Betaseron, and outside the US as Betaferon.
=== Mechanistic vs. empirical models === A main difference in evolutionary models is how many parameters are estimated every time for the data set under consideration and how many of them are estimated once on a large data set. Mechanistic models describe all substitutions as a function of a number of parameters which are estimated for every data set analyzed, preferably using maximum likelihood. This has the advantage that the model can be adjusted to the particularities of a specific data set (e.g. different composition biases in DNA). Problems can arise when too many parameters are used, particularly if they can compensate for each other (this can lead to non-identifiability). Then it is often the case that the data set is too small to yield enough information to estimate all parameters accurately. Empirical models are created by estimating many parameters (typically all entries of the rate matrix as well as the character frequencies, see the GTR model above) from a large data set. These parameters are then fixed and will be reused for every data set. This has the advantage that those parameters can be estimated more accurately. Normally, it is not possible to estimate all entries of the substitution matrix from the current data set only. On the downside, the parameters estimated from the training data might be too generic and therefore have a poor fit to any particular dataset. A potential solution for that problem is to estimate some parameters from the data using maximum likelihood (or some other method).
Sources: en.wikipedia.org
Marion Walwyn, a cousin who first met Banting in 1901, recalled that "we sat together in the swing in our yard. In an hour he didn't say one word." He continued to struggle in school and stubbornly resisted being disciplined there. After one incident, he resolved never to continue his education but was convinced otherwise by his father. Banting's grandfather, John Banting, had urged his own children to be educated; the philosophy had influenced William, who offered to provide a fund to his sons when they turned twenty-one. In contrast to his brothers, who spent the inheritance towards their own farms, Frederick would use it towards matriculation. In his late teenage years, Banting grew into a tall man with engagements in school football and baseball teams. Both his mother and father hoped that he would find a vocation in the Methodist ministry. He passed physics and chemistry during junior matriculation examinations in 1909, but repeated English and was required to undertake French and Latin. The next year, he narrowly passed Latin but failed French and, for a second time, English composition. The principal later remembered his repeated efforts: "We would not have picked him for one on whom fame should settle. He was a white boy, a right boy."
== Football career == He made his debut in 2001 with the Essendon Football Club and proved to be a solid ruckman for a number of years. In 2006, Hille was appointed stand-in captain after Matthew Lloyd suffered a season-ending hamstring injury in round three. He captained his first game in Round 6, 2006. From 2007 to 2012, he held the role of vice-captain of the Essendon Football Club which he shared with Mark McVeigh for the most part. In the Round 17, 2006 clash between Essendon and Brisbane Lions (the first match Essendon won under his captaincy, and since the beginning of April that year), Brisbane ruckman Jamie Charman fell heavily in a contest, seriously injuring his shoulder. The image of David Hille comforting his opponent whilst medical help arrived was often replayed by the football media as a rare example of sportsmanship in such a competitive game. At the 2008 Essendon Best and Fairest, Hille swept the floor by taking the most prestigious award of the night, the 'Best and Fairest'. Hille also came away with the 'Cheersquad award'. Hille suffered a knee injury in the second minute of the Bombers' annual ANZAC Day match against Collingwood in Round 5, 2009. Scans confirmed he had ruptured his anterior cruciate ligament, requiring a full knee construction and ending his season.
==== Molecular interactions ==== As MALS can provide molar mass and size of molecules, it permits study into protein-protein binding, oligomerization and the kinetics of self-assembly, association and dissociation. By comparing the molar mass of a sample to its concentration, one can determine the binding affinity and stoichiometry of interacting molecules.
Sources: en.wikipedia.org
It separates components in a liquid sample and measures their amounts using a detector. Results can indicate concentration, purity, or identity based on retention time and detector response. The technique works for mixtures that can be dissolved and filtered.
It offers high resolution, reproducibility, and compatibility with many sample types. A single run can separate and quantify multiple analytes. It is common in pharmaceutical, food, environmental, and industrial laboratories.
Samples must be soluble in a suitable mobile phase and free of particles that can block the column. Detector response depends on analyte structure, so some compounds need derivatization or alternative detection. Complex matrices may require extensive sample preparation.
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.