retention time raises a handful of sensible questions. This page answers them in order, starting with the fundamentals and moving to applications.
This page was last updated on 2026-04-04 and is reviewed periodically as new material appears.
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.
Routine quality control uses system suitability, blank injections, check standards, and control samples to detect drift or contamination. System suitability criteria may specify minimum resolution, maximum tailing factor, and a permitted range for repeated injections. Blank injections reveal carryover or solvent contamination, while check standards confirm calibration accuracy over a batch. Control samples with known analyte levels can show whether results remain within statistical limits. When a control result falls outside limits, the analyst investigates the cause and may invalidate affected results before repeating the batch.
Documentation and traceability are central to regulated HPLC testing. Records typically include instrument logs, column history, mobile-phase preparation, sample preparation, injection sequences, raw chromatograms, and audit trails. Electronic systems may require user access controls, time-stamped changes, and backup procedures. Training records show that analysts are qualified for assigned methods. Audits and inspections check whether written procedures match actual practice and whether deviations are documented. These controls support reproducibility and allow results to be reconstructed if questions arise later.
| 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 |
Key performance measures include retention time, peak area, peak height, resolution, tailing factor, and plate count. Retention time helps identify a peak under fixed conditions, but confirmation often requires a second method or detector. Peak area and height relate to concentration through calibration curves, which may be linear or nonlinear depending on the detector response. Resolution describes separation between adjacent peaks, while tailing factor and plate count describe peak shape and column efficiency. Performance checks verify these values before and during a run to confirm that the instrument is performing within limits.
High-performance liquid chromatography testing separates components of a liquid sample by forcing a mobile phase through a packed column. The stationary phase inside the column interacts with analytes to different degrees, so each compound exits at a characteristic retention time. A pump delivers solvent at controlled flow and pressure, while an injector introduces a precise sample volume. Detectors such as ultraviolet-visible, fluorescence, refractive index, or mass spectrometric instruments record the separated bands. The resulting chromatogram provides qualitative and quantitative information about the mixture.
Separation modes differ by the chemistry of the stationary phase and the composition of the mobile phase. Reversed-phase testing uses a nonpolar column and polar solvents, making it common for pharmaceutical, environmental, and food analytes. Normal-phase testing uses a polar column and nonpolar solvents for compounds that are poorly retained in reversed-phase systems. Ion-exchange and ion-pair methods separate charged species, while size-exclusion methods sort molecules by hydrodynamic volume. Gradient elution changes solvent strength over time to resolve complex mixtures, and isocratic elution holds solvent composition constant for simpler assays.
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.
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.
When fomepizole is not available, ethanol can be used to treat or prevent methanol or ethylene glycol poisoning. The rate-limiting steps for the elimination of ethanol are in common with these substances, so it competes with other alcohols for the alcohol dehydrogenase enzyme. Methanol itself is not highly toxic, but its metabolites formaldehyde and formic acid are; therefore, to reduce the rate of production and concentration of these harmful metabolites, ethanol can be ingested or injected. This avoids the production of toxic aldehyde and carboxylic acid derivatives, and reduces the more serious toxic effects of the glycols when crystallized in the kidneys. Ethylene glycol poisoning can be treated in the same way.
The very success of modernization encourages "utopian" hopes that all problems can be solved via the "spirit of science" that are inevitably dashed. Modern society causes unavoidable "irritations" which led to people looking backwards to "traditions" and/or a "clean" modernity where the state would attempt to solve social problems via radical means. The "demystification of the world" leads people to seek faith and self-validation either via irrational theories such as "race" and/or a charismatic leader who would revitalize society. Modernity creates a mass society that can be more easily manipulated and mobilized to ends that can be either moral or amoral. Peukert argued that starting in 1929 that the disjoint between Weimar democracy vs. the problems of "classical modernity" started to fell apart when faced with the Great Depression. Peukert maintained that the Weimar Republic was a muddled system built out of the compromises between so many different interests with for instance Weimar Coalition consisting of the left-wing SPD, the liberal DDP, and the centre-right Zentrum being the only political parties wholeheartedly committed to the Weimar republic. Other competing interests in Germany included the struggle between men vs. women, farmers vs. towns, Catholics vs. Protestants, and unions vs. business.
=== Less common causes === In primary biliary cholangitis (previously known as primary biliary cirrhosis), the bile ducts become damaged by an autoimmune process. This leads to liver damage. Some people may have no symptoms, while others may present with fatigue, pruritus, or skin hyperpigmentation. The liver is typically enlarged which is referred to as hepatomegaly. Rises in alkaline phosphatase, cholesterol, and bilirubin levels occur. Patients are usually positive for anti-mitochondrial antibodies. Primary sclerosing cholangitis is a disorder of the bile ducts that presents with pruritus, steatorrhea, fat-soluble vitamin deficiencies, and metabolic bone disease. A strong association with inflammatory bowel disease is seen, especially ulcerative colitis. Autoimmune hepatitis is caused by an attack of the liver by lymphocytes. This causes inflammation and eventually scarring as well as cirrhosis. Findings include elevations in serum globulins, especially gamma globulins. Hereditary hemochromatosis usually presents with skin hyperpigmentation, diabetes mellitus, pseudogout, or cardiomyopathy. All of these are due to signs of iron overload. Family history of cirrhosis is common as well. Wilson's disease is an autosomal recessive disorder characterized by low ceruloplasmin in the blood and increased copper of the liver. Copper in the urine is also elevated. People with Wilson's disease may also have Kayser–Fleischer rings in the cornea and altered mental status.
Progesterone is marketed under a large number of brand names throughout the world. Examples of major brand names under which progesterone has been marketed include Crinone, Crinone 8%, Cyclogest, Endogest, Endometrin, Estima, Geslutin, Gesterol, Gestone, Luteina, Luteinol, Lutigest, Lutinus, Microgest, Progeffik, Progelan, Progendo, Progering, Progest, Progestaject, Progestan, Progesterone, Progestin, Progestogel, Prolutex, Proluton, Prometrium, Prontogest, Strone, Susten, Utrogest, and Utrogestan.
Sources: en.wikipedia.org
Ohnologous genes are paralogous genes that have originated by a process of whole-genome duplication. The name was first given in honour of Susumu Ohno by Ken Wolfe. Ohnologues are useful for evolutionary analysis because all ohnologues in a genome have been diverging for the same length of time (since their common origin in the whole genome duplication). Ohnologues are also known to show greater association with cancers, dominant genetic disorders, and pathogenic copy number variations.
== Pharmacokinetics == Abacavir is given orally and is rapidly absorbed with a high bioavailability of 83%. Solution and tablet have comparable concentrations and bioavailability. Abacavir can be taken with or without food. Abacavir can cross the blood–brain barrier. Abacavir is metabolized primarily through the enzymes alcohol dehydrogenase and glucuronyl transferase to an inactive carboxylate and glucuronide metabolites. It has a half-life of approximately 1.5-2.0 hours. If a person has liver failure, abacavir's half-life is increased by 58%. Abacavir is eliminated via excretion in the urine (83%) and feces (16%). It is unclear whether abacavir can be removed by hemodialysis or peritoneal dialysis.
Valve was named as a defendant in two lawsuits in June and July 2016 related to third-party gambling sites that use the Steamworks API to allow betting with the virtual currency of cosmetic weapon replacement textures, better known as "skins", from Counter-Strike: Global Offensive, which through these sites can be converted from or to real-world money. Both suits assert Valve aiding in underaged gambling. Valve subsequently stated it has no commercial ties with these sites, and that it would demand these sites cease their use of the Steamworks API as they violate the authorized use policies. In October 2016, the Washington State Gambling Commission required Valve to stop the use of virtual skins for gambling on Steam, stating they would face legal repercussions if they failed to co-operate. On October 17, 2016, Valve sent a letter to the Washington State Gambling Commission stating that it had "no business relationship with such gambling sites", asserting that they come into existence, operate, and go out of existence without its knowledge and consent, adding that it was not aware of any such law that Steam or any of its games were violating.
==== Absorption and bioavailability ==== The oral bioavailability of estradiol is very low. This is due to the fact that estradiol is poorly soluble in water, which limits its dissolution and absorption, and is additionally subject to extensive metabolism during the first pass through the intestines and liver. Estradiol is micronized and/or conjugated with an ester, as in estradiol valerate or estradiol acetate, to improve its oral bioavailability and potency. Micronization decreases the particle size of estradiol crystals and hence increases the surface area for absorption, thereby improving the rate and extent of absorption. In addition, there is an improvement in metabolic stability. Oral micronized estradiol consists of more than 80% of estradiol particles micronized to a size smaller than 20 μm in diameter, or to about 1 to 3 μm on average. All oral formulations of estradiol available today are micronized, and oral estradiol valerate tablets also seem to be micronized. Oral non-micronized estradiol and oral micronized estradiol do not appear to have ever been directly compared in a study. Both have been assessed independently however, and have been found to produce significant estrogenic effects. Micronization of other poorly water-soluble steroids such as spironolactone and norethisterone acetate has been found to increase their potency by several-fold.
The androgen backdoor pathways are vital for creating androgens from 21-carbon (C21) steroids, known as pregnanes. A 21-carbon steroid is a steroid molecule with 21 carbon atoms, hence, their chemical formula contains C21. For example, the chemical formula of progesterone is C21H30O2. For this reason, 21-carbon steroids are denoted as C21-steroids, 19-carbon steroids are denoted as C19 steroids, and so on. The androgen backdoor pathways occur without the involvement of testosterone (T) and/or androstenedione (A4), which are part of the conventional, canonical (classic) androgenic pathway. In the canonical pathways of androgen biosynthesis, DHT is synthesized from T via 5α-reduction, so that 5α-reduction of T, a C19 steroid, is the last step of the pathway (see Dihydrotestosterone § Biosynthesis). In the backdoor pathways, to the contrary, 5α-reduction of C21 steroids is the first step. The 5α-reduction is a chemical reaction where a functional group attached to the carbon in position 5α of the steroid nucleus is reduced, and a double bond between carbon atoms numbered 4 and 5 in the steroid molecule is replaced to the single bond in a chemical reaction catalyzed by the SRD5A1 enzyme. The androgen backdoor pathways can be also activated in pathologic conditions (diseases), such as congenital adrenal hyperplasia (CAH), leading to hyperandrogenism.
Sources: en.wikipedia.org
Blood smear examination is usually performed in conjunction with a complete blood count in order to investigate abnormal results or confirm results that the automated analyzer has flagged as unreliable. Microscopic examination of the shape, size, and coloration of red blood cells is useful for determining the cause of anemia. Disorders such as iron deficiency anemia, sickle cell anemia, megaloblastic anemia and microangiopathic hemolytic anemia result in characteristic abnormalities on the blood film. The proportions of different types of white blood cells can be determined from the blood smear. This is known as a manual white blood cell differential. The white blood cell differential can reveal abnormalities in the proportions of white blood cell types, such as neutrophilia and eosinophilia, as well as the presence of abnormal cells such as the circulating blast cells seen in acute leukemia. Qualitative abnormalities of white blood cells, like toxic granulation, are also visible on the blood smear. Modern complete blood count analyzers can provide an automated white blood cell differential, but they have a limited ability to differentiate immature and abnormal cells, so manual examination of the blood smear is frequently indicated. Blood smear examination is the preferred diagnostic method for certain parasitic infections, such as malaria and babesiosis. Rarely, bacteria may be visible on the blood smear in patients with severe sepsis.
These results highlight the clinical translation of α-amanitin’s cell-cycle-independent mechanism, which effectively targets both proliferating tumor cells and quiescent cancer stem cells. The ATAC platform is also advancing into solid tumors. TROP2-targeted conjugates based on the hRS7 antibody achieved complete tumor eradication in patient-derived xenograft models of pancreatic cancer and triple-negative breast cancer, outperforming the approved ADC sacituzumab govitecan. In January 2026, Takeda initiated a Phase I/II clinical trial of an ATAC candidate for solid tumors.
In mammals, the genome only contains one gene for tropoelastin, called ELN. The human ELN gene is a 45 kb segment on chromosome 7, and has 34 exons interrupted by almost 700 introns, with the first exon being a signal peptide assigning its extracellular localization. The large number of introns suggests that genetic recombination may contribute to the instability of the gene, leading to diseases such as SVAS. The expression of tropoelastin mRNA is highly regulated under at least eight different transcription start sites. Tissue specific variants of elastin are produced by alternative splicing of the tropoelastin gene. There are at least 11 known human tropoelastin isoforms. These isoforms are under developmental regulation, however there are minimal differences among tissues at the same developmental stage.
In 2015, in the Journal of Molluscan Studies, Puillandre, Duda, Meyer, Olivera & Bouchet presented a new classification for the old genus Conus. Using 329 species, the authors carried out molecular phylogenetic analyses. The results suggested that the authors should place all cone snails in a single family, Conidae, containing four genera: Conus, Conasprella, Profundiconus and Californiconus. The authors group 85% of all known cone snail species under Conus. They recognize 57 subgenera within Conus, and 11 subgenera within the genus Conasprella.
Botulinum toxin, botulinum neurotoxin, or botox® is a neurotoxic protein produced by the bacterium Clostridium botulinum and related species, and it is considered the deadliest known natural substance ever recorded in chemical literature. It prevents the release of the neurotransmitter acetylcholine from axon endings at the neuromuscular junction, thus causing flaccid paralysis. The toxin causes the disease botulism. Botulinum toxin is an acetylcholine release inhibitor and a neuromuscular blocking agent. Botulinum toxin was developed as a biological agent by the Soviet, United States, and Iraqi biological weapons programs. In strongly diluted form, the toxin is also used for medical and cosmetic purposes. The seven main types of botulinum toxin are named types A to G (A, B, C1, C2, D, E, F and G). New types are occasionally found. Types A and B are capable of causing disease in humans, and are also used commercially and medically. Types C–G are less common. In 2025, the structure of the complete 14 subunit botulinum neurotoxin complex (L-PTC) was solved.
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.
Validation establishes suitability for a new method, while verification confirms that a method works in a specific laboratory. Verification is often used when a validated method is adopted with existing equipment and staff. Both rely on documented acceptance criteria.