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 2025-07-30. Where a claim depends on a specific study, the study is described rather than over-claimed.
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, or HPLC, separates dissolved compounds by passing a liquid mobile phase through a packed column. Components distribute differently between the stationary phase and the moving liquid, so they travel at different speeds and exit at different times. A detector records these eluting bands as peaks, and peak area or height relates to amount. The technique supports testing in pharmaceuticals, foods, environmental samples, and industrial chemicals. Quantification usually depends on calibration with known standards.
High-performance liquid chromatography is an analytical technique that separates components in a liquid sample by passing them through a packed column under pressure. A pump delivers a mobile phase at a controlled flow rate, and an injector introduces the sample into the stream. Differences in how analytes partition between the mobile phase and the stationary phase cause them to exit the column at different times. Detection then records a signal proportional to the amount of each separated substance. The resulting chromatogram provides retention times and peak areas for identification and quantification.
Instrumentation includes a solvent delivery system, an autosampler, a column oven, and one or more detectors. Reversed-phase columns with chemically modified silica are widely used, but normal-phase, ion-exchange, size-exclusion, and affinity modes exist for specific separations. Detectors may rely on ultraviolet absorbance, fluorescence, refractive index, or mass spectrometry. Column temperature, mobile phase composition, and flow rate are adjusted to improve resolution. System pressure is monitored because rising pressure can indicate column blockage or deteriorating packing.
| 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. |
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.
HPLC testing is not a single fixed procedure; it is a family of separation modes. Reversed-phase, normal-phase, ion-exchange, size-exclusion, and affinity chromatography each suit different analyte properties. Reversed-phase methods dominate because they handle many neutral and moderately polar compounds. Detection can be optical, electrochemical, or mass spectrometric, and the detector dictates what information is available. Coupling with mass spectrometry increases selectivity and enables identification when standards are unavailable. The technique cannot separate every mixture without adjustment.
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.
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.
High-performance liquid chromatography is an analytical technique that separates components of a liquid sample by passing it through a packed column under pressure. A pump delivers a mobile phase at a controlled flow rate, and an injector introduces the sample into the stream. Differences in interaction with the stationary phase cause analytes to migrate at different rates. Detectors record elution as peaks, and a data system converts signals into a chromatogram. The method is suited to compounds that dissolve in a liquid and are not volatile enough for gas chromatography.
Reversed-phase chromatography dominates modern HPLC testing, using a nonpolar stationary phase such as chemically bonded octadecyl groups and a polar mobile phase of water mixed with organic solvent. Analytes partition between the mobile and stationary phases according to hydrophobicity. Gradient elution changes the mobile phase composition over time to separate compounds with a wide range of retention. Isocratic elution keeps the composition constant and is simpler for routine assays. Column temperature, pH, and flow rate influence selectivity, peak shape, and retention time, so these parameters are controlled during a validated method.
Much of the literature, however, suggests a curvilinear or even quadratic relationship between spatial performance and circulating testosterone, where both hypo- and hypersecretion (deficient- and excessive-secretion) of circulating androgens have negative effects on cognition.
=== Drug repurposing === Chemoproteomics is at the forefront of drug repurposing. This is particularly relevant in the era of COVID-19, which saw a dire need to rapidly identify FDA approved drugs that have antiviral activity. In this context, a phenotypic screen is usually employed to identify drugs with a desired effect in vitro, such as inhibition of viral plaque formation. If a drug produces a positive test, the next step is to determine whether it is acting on a known or novel target. Chemoproteomics is thus a follow-up to phenotypic screening. In the case of COVID-19, Friman et al investigated off-target effects of the broad-spectrum antiviral Remdesivir, which was among the first repurposed drugs to be used in the pandemic. Remdesivir was tested via thermal proteome profiling in a HepG2 cellular thermal shift assay, along with the controversial drug hydroxychloroquine, and investigators discovered TRIP13 as a potential off-target of Remdesivir.
From the same toxin family of bungarotoxins, κ-BTX was shown to act postsynaptically on α3 and α4 neuronal nicotinic receptors with little effect on the muscular nAChRs, targeted by α-BTX. In contrast, β- and γ-BTX act presynaptically by reducing ACh release. Neurotoxins are named based on the receptor type they target. The nicotinic receptors are made up of five subunits each and contain two binding sites for snake venom neurotoxins. The α7-nAChR is a homopentamer consisting of five identical α7 subunits. The α7 receptor is known to have a higher Ca2+ permeability compared to other nicotinic receptors. Changes in Ca2+ intracellularly can activate important cellular pathways such as the STAT pathway or the NF-κB signalling. Consistency with experimental data on the amount of toxin per receptor is evident in the observation that a lone molecule of the toxin is adequate to inhibit channel opening. Some computational studies of the mechanism of inhibition using normal mode dynamics suggest that a twist-like motion caused by ACh binding may be responsible for pore opening and that this motion is inhibited by toxin binding.
Sources: en.wikipedia.org
==== Tax credit for seniors ==== The law permanently eliminates the personal exemption, which had been temporarily eliminated by the Tax Cuts and Jobs Act of 2017. It offers a temporary tax deduction, set to expire in 2028, of up to $6,000 for seniors. The deduction phases out for individuals with modified adjusted gross income (MAGI) exceeding $75,000 (or $150,000 for married couples). According to the Council of Economic Advisors, this would result in 88% of seniors being able to claim enough deductions to clear their Social Security tax burden, up from 64% under prior law. A taxpayer is not required to itemize their tax deductions in order to take the tax deduction.
==== Film criticism ==== A significant number of users on TikTok, such as Juju Green, create content surrounding film criticism and easter eggs. However, as reported by The New York Times, these people often do not see themselves necessarily as film critics. These creators would often attend red carpet premieres of movies and interview the celebrities in attendance, which was the subject of significant debate as some considered the questions the creators asked to be disrespectful. By 2022, TikTok released a Showbiz List, highlighting individuals who were having a larger impact on the film industry. During the 2023 SAG-AFTRA strike, such influencers were told that they would be denied future entry into the union if they partnered with struck studios. This led many creators to stop creating new content which they were not already contractually obligated to create. Creators who posted saying that they would not be changing their content, such as Green, were met with significant criticism.
The prefix bi- (for joining of ring systems) is translated as 联 (lián, 'join', 'couple'), as in 联苯 for biphenyl. The stereochemical descriptors cis- and trans- are translated as 顺 (shùn, 'along') and 反 (fǎn, 'against'). The relational prefixes ortho-, meta-, and para- are translated as 邻 (lín, 'neighboring'), 间 (jiàn, 'between'), and 对 (duì, 'opposing'), respectively. The structural modification descriptors cyclo-, nor- and homo- are translated as 环 (huán, 'ring'), 降 (jiàng, 'lowered'), and 高 (gāo, 'high'). For example, norbornene is translated as 降冰片烯, in which the trivial name (冰片) for bornyl [literally, camphor] is used. When substitutive nomenclature is used for naming heterocycles, the suffix 杂 (zá, 'mixed') is used in the same way as '-a' in English (as in aza, thia, oxa, etc.). As an example, DABCO (1,4-diazabicylo[2.2.2]octane) is named 1,4-二氮杂二环［2.2.2］辛烷. The common unsaturated groups allyl and propargyl are translated as 烯丙(基) (xībǐng(jī), 'alkene-prop-(yl)') and 炔丙(基) (qūebǐng(jī), 'alkyne-prop-(yl)'). Thus, using 高 for homo- and 烯丙 for allyl, 3-buten-1-ol is also called 高烯丙醇 (i.e., homoallyl alcohol) in Chinese via semisystematic nomenclature. The Chinese Wikipedia page may be consulted for further details.
Sources: en.wikipedia.org
The following is a list of fentanyl analogues (sometimes referred to as fentalogs), and includes both compounds developed by pharmaceutical companies for legitimate medical use, and those which have been sold as designer drugs. The latter have been reported to national drug control agencies such as the DEA, and some to transnational agencies such as the EMCDDA and UNODC. This is not a comprehensive or exhaustive list of fentanyl analogues, as more than 1400 compounds from this family have been described in the scientific and patent literature. However, this list does include many notable compounds that have reached late-stage human clinical trials, and compounds which have been sold as designer drugs, as well as representative examples of significant structural variations reported in the scientific and patent literature. The structural variations among fentanyl analogues can impart profound pharmacological differences between each other, especially regarding potency and efficacy. In the United States, the Drug Enforcement Administration (DEA) placed the class of "Fentanyl-Related Substances" on the list of Schedule I drugs in 2018, making it illegal to manufacture, distribute, or possess fentanyl analogs, with very broad terminology being used in its scheduling. Regarding the temporary control of fentanyl-related substances, Schedule I was extended through December 31, 2024 by Public Law 117-328.
=== Biological === An MRI study conducted on Price provides a plausible argument as to the neurological foundation of her superior memory. Both the temporal lobe and the caudate nucleus were found to be enlarged. Parker and colleagues speculated that a defective frontostriatal circuit could be responsible for the observed executive function deficits in hyperthymesia. This circuit plays a crucial role in some neurodevelopmental disorders including obsessive–compulsive disorder and Alzheimer's. Given the parallels in some aspects of behavior, Price's hyperthymestic abilities possibly stem from atypical neurodevelopment. Scientists now need to ascertain if and how these brain areas are connected to establish a coherent neurological model for superior autobiographical memory. For autobiographical memory, the hippocampus, located in the medial temporal lobe, is involved in the encoding of declarative memory (memory for facts and events), while the temporal cortex is involved in the storage of such memory. The caudate nucleus is primarily associated with procedural memory, in particular habit formation, and is, therefore, intrinsically linked to obsessive-compulsive disorder. A 2018 clinical trial published that there were higher levels of activation in the medial prefrontal cortex and temporoparietal junction along with heightened connection between the prefrontal cortex and the hippocampus in hyperthymesiacs, suggesting that these regions may play a role in the enablement of the condition.
Often rules apply to all goods neutrally, but may have a greater practical effect on imports than domestic products. For such "indirect" discriminatory (or "indistinctly applicable") measures the Court of Justice has developed more justifications: either those in article 36, or additional "mandatory" or "overriding" requirements such as consumer protection, improving labour standards, protecting the environment, press diversity, fairness in commerce, and more: the categories are not closed. In the noted case Rewe-Zentral AG v Bundesmonopol für Branntwein, the Court of Justice found that a German law requiring all spirits and liqueurs (not just imported ones) to have a minimum alcohol content of 25 per cent was contrary to TFEU article 34, because it had a greater negative effect on imports. German liqueurs were over 25 per cent alcohol, but Cassis de Dijon, which Rewe-Zentrale AG wished to import from France, only had 15 to 20 per cent alcohol. The Court of Justice rejected the German government's arguments that the measure proportionately protected public health under TFEU article 36, because stronger beverages were available and adequate labelling would be enough for consumers to understand what they bought. This rule primarily applies to requirements about a product's content or packaging. In Walter Rau Lebensmittelwerke v De Smedt PVBA the Court of Justice found that a Belgian law requiring all margarine to be in cube shaped packages infringed article 34, and was not justified by the pursuit of consumer protection.
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 separates and detects individual compounds in a liquid sample, producing peaks at characteristic retention times. Peak area or height can be used to estimate concentration when calibrated with known standards. It does not identify unknown compounds with certainty unless additional detectors or reference materials are used.