Perchlorate

HPLC Method for Separation and Analysis of Acidic Toxin Metabolites with an MS-Compatible Mobile Phase on a Newcrom B Column by SIELC Technologies.

Separation type: Liquid Chromatography Mixed-mode

Metabolites and biomarkers are key terms in the fields of biochemistry, pharmacology, and medicine.

1. Metabolites:
   • Definition: These are small molecules produced during metabolism, which is the set of chemical reactions that occur within cells to maintain life. Metabolites can be either products of metabolism or the substrates used in metabolic reactions.
   • Types: There are primary metabolites, which are directly involved in normal growth, development, and reproduction (like amino acids and sugars), and secondary metabolites, which often have specialized roles such as defense mechanisms in plants (like alkaloids or antibiotics).
   • Uses: In the context of drug testing, for instance, the presence of certain metabolites can reveal whether someone has taken a specific drug, even if the drug itself is no longer present in the person’s system.
2. Biomarkers:
   • Definition: A biomarker, or biological marker, is a measurable indicator of some biological state or condition. Biomarkers are often used to assess the risk or presence of disease, to monitor the progress of disease, or to see how the body responds to a treatment.
   • Types: Biomarkers can be specific cells, molecules, genes, enzymes, hormones, or metabolites. They can also be measurements like heart rate or blood pressure.
   • Uses: In medicine, biomarkers can be used for early disease detection (like elevated PSA levels indicating potential prostate cancer), monitoring disease progression (like HIV viral load tests to see how well antiretroviral treatment is working), or understanding disease mechanisms. They can also be used to see if a body has been exposed to certain harmful substances, indicating potential harm or risk.

N-Acetyl-S-(3-hydroxypropyl)-L-cysteine (3HPMA) is a common metabolite of the herbicide and pollutant Acrolein. 2-Hydroxyisobutyric Acid (2HIB) is a neuro and adrenal toxin that is a common metabolite of the fuel additives Methyl tert-butyl ether (MTBE) and Ethyl tert-butyl ether (ETBE). N-Acetyl(Carbomylethyl)cysteine (NAE) is a metabolite of Acrylamide which can act as a neuro toxin in cases of long term exposure. N-Acetyl(propyl)cysteine (NAPR) is a metabolite of the popular industrial organic solvent 1-bromopropane (1-BP). Perchlorate (PERC) inhibits thyroid activity and can be used to treat hyperthyroidism, but can be toxic in large amounts or in regularly-functioning thyroids. Diphenylphosphate (DPP) is a metabolite of the popular flame retardant triphenyl phosphate (TPHP). These acidic toxin metabolites can be retained, separated, and analyzed on a mixed-mode Newcrom B column with a mobile phase consisting of water, Acetonitrile (MeCN), and Ammonium Acetate (AmAc). This analytical method can be detected with high resolution and peak symmetry with many evaporative detection methods, including Evaporative Light Scattering Detection (ELSD), Charged Aerosol Detector (CAD), and Electrospray Ionization (ESI) for Mass Spectrometry (MS).

High Performance Liquid Chromatography (HPLC) Method for Analyses of Michler’s ketone

Condition

Description

Pesticide is a more generic term that includes herbicides, fungicides and insecticides in its definition. Herbicides are used to control unwanted plants, they are also known as weedkillers. Insecticides are used to kill insects. Fungicides are used to kill parasitic fungi. All are heavily used in agriculture. By using HPLC, many different pesticides can be separated and their retention characteristics controlled using the Newcrom B mixed-mode column.

The ionic forms of Chlorate, Perchlorate, and Phosphonates are useful in many industries including medicine, paper and use in explosives. Due to their lack of UV activity, an ELSD was used to detect both the anions and cations of all three sodium salts. The ions were retained on both Primesep D and Obelisc R columns. Primesep D is a reverse phase column with embedded basic ion-pairing groups. Obelisc R is also a reverse phase column, but can be additionally tuned due to embedded ionic groups and a hydrophobic chain.

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Main source of contamination of environment by perchlorates are rocket fuels, car airbags, and fireworks.
EPA developed regulation of the perchlorate level in drinking water which already adapted by several states.
The most convenient, universal and very sensitive way to measure perchlorate is chromatography.
SIELC developed a simple, rugged, and selective HPLC method which allows to measure perchlorate in different matrices including drinking water.
This method allows to measure simultaneously other chloro containing ions such as chloride, and chlorate.
The method shows high selectivity and specificity.
The mobile phase is a simple mixture of water acetonitrile and ammonium formate.
The column used in the analysis has advanced surface chemistry with long chain holding a terminal positively changed functional group.

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The ionic forms of Chlorate, Perchlorate, and Phosphonates are useful in many industries including medicine, paper and use in explosives. Due to their lack of UV activity, an ELSD was used to detect both the anions and cations of all three sodium salts. The ions were retained on both Primesep D and Obelisc R columns. Primesep D is a reverse phase column with embedded basic ion-pairing groups. Obelisc R is also a reverse phase column, but can be additionally tuned due to embedded ionic groups and a hydrophobic chain.

The majority of drugs in the pharmaceutical industry are administered in salt form. The presence of two counter-ions very often necessitates the use of two methods. The nature of these counterparts in drugs can be an inorganic cation and organic acid, inorganic anion and organic base, and organic cation and organic anion. Furthermore, the properties of the molecules will result in a differing stoichiometry. The task of simultaneous quantitation of counter-ions can be achieved by using mixed-mode columns. The general approach for analysis is based on properties of corresponding counter-ions. Hydrophobic basic drugs, like dextromethorphan, verapamil, trimipramine, and corresponding acidic counter-ions (chloride, chlorate, bromide, bromate, perchlorate, maleate, fumarate,tartrate, succinate, phosphate, citrate, benzosulfonate, toleuensulfonate) can be separated and quantitated in the same run on reversed-phase anion-exchange column. Basic hydrophobic drugs are retained by the reversed-phase mechanism, and counter-ions are retained by the reversed-phase and anion-exchange mechanism. Some polar counter-ions are retained only by the anion-exchange mechanism. Retention time and selectivity of HPLC separation of drugs and counter-ions can be achieved by changing the amount of acetonitrile and the amount of ions in the mobile phase. The detection technique depends on the properties of the counter-ions. In case of low or no UV activity, ELSD can be employed if the counter-ion forms a non-volatile salt with the mobile phase additive (ammonium formate). This HPLC method can be used for simultaneous quantitation of other basic drugs and counter-ions. The presence of two mechanisms of retention allows control over retention times of drug and counter-ion independently, and even allows a change of order of elution when necessary.