HPLC-MS Method for Separation of Metabolites and Biomarkers on Newcrom B Column

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

HPLC-MS Method for Separation of Metabolites and Biomarkers on Newcrom B Column by SIELC Technologies
HPLC Method for Analysis of Metabolites and Biomarkers on Newcrom B Column by SIELC Technologies

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


ColumnNewcrom B, 4.6 x 150 mm, 5 µm, 100 A
Mobile PhaseGradient MeCN/H2O – 5-50%, 20 min
BufferGradient Ammonium acetate pH 4.0 –10-25 mM, 20 min
Flow Rate1.0 ml/min
DetectionESI- and ESI


Class of CompoundsMetabolites and Biomarkers
Analyzing CompoundsMetabolites and Biomarkers

Application Column

Newcrom B

Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:
2-Hydroxyisobutyric acid
Diphenyl phosphate
N-Acetyl (Carbamoylmethyl) cysteine
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.