Hypoxanthine

HPLC Method for Analysis of Uracil, Thymine, Hypoxanthine, Guanine on Newcrom AH Column by SIELC Technologies

Separation type: Liquid Chromatography Mixed-mode

Uracil, Thymine, Hypoxanthine, and Guanine are all nitrogenous bases, each playing distinct roles in the biochemistry of nucleic acids.

Uracil (U):

• Found in: RNA
• Pairs with: Adenine (A)
• Structure: Pyrimidine
• Role: Uracil replaces thymine in RNA. In DNA, adenine pairs with thymine, but in RNA, adenine pairs with uracil.

Thymine (T):

• Found in: DNA
• Pairs with: Adenine (A)
• Structure: Pyrimidine
• Role: Thymine is specific to DNA, distinguishing it from RNA. It’s the base that pairs with adenine through two hydrogen bonds.

Hypoxanthine:

• A naturally occurring purine derivative. It’s not directly a base in standard DNA or RNA, but it plays a significant role in the metabolism of purines.
• It is the base form of the nucleoside inosine, which can be found in certain tRNAs and plays a role in wobble base pairing.
• Hypoxanthine is also an intermediate in the purine degradation pathway, leading to the production of uric acid.

Guanine (G):

• Found in: Both DNA and RNA
• Pairs with: Cytosine (C)
• Structure: Purine
• Role: Guanine is one of the four main nucleobases in the nucleic acids DNA and RNA. It forms three hydrogen bonds with cytosine, contributing to the stability of the nucleic acid structures.

These nitrogenous bases are crucial for the structure, replication, and function of nucleic acids. Understanding their properties and interactions is fundamental to molecular biology and genetics.

Nucleotides can be retained and analyzed on a mixed-mode Newcrom AH column with a mobile phase consisting of water, Acetonitrile (MeCN), and ammonium formate. This analytical method can detect compounds with high resolution and peak symmetry using UV detection at 260 nm

High Performance Liquid Chromatography (HPLC) Method for Analyses of Uracil, Thymine, Hypoxanthine and Guanine

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HPLC Method for Analysis mixture of Xanthines and Uric Acid BIST B+ by SIELC Technologies.

Separation type: Hydrophilic interaction liquid chromatography (HILIC)

Xanthines and uric acid are related compounds in the body and both are involved in the metabolism of purines.

Xanthines are a group of alkaloids that are widely distributed in plants, and also occur in the tissues and fluids of animals. They are known to stimulate the central nervous system and cardiac muscle, and also have diuretic effects. Some commonly known xanthines include caffeine (found in coffee, tea, and chocolate), theobromine (found in cocoa and chocolate), and theophylline (used as a drug in the treatment of respiratory diseases like asthma).

In the body, xanthines are intermediates in the degradation of adenosine monophosphate to uric acid. This metabolic pathway starts with adenosine monophosphate (AMP), which is deaminated to form inosine monophosphate (IMP). IMP is then converted into a xanthine known as hypoxanthine. Hypoxanthine is then oxidized to xanthine, and finally, xanthine is further oxidized to uric acid. Both of the oxidation steps are catalyzed by the enzyme xanthine oxidase.

Uric acid and Xanthines can be retained, analyzed, and separated using an isocratic analytical method on a BIST B+ column. The simple mobile phase for this method comprises water, acetonitrile (MeCN), and formic acid as an ionic modifier. The analytical method can be monitored with UV detection at 260 nm, an Evaporative Light Scattering Detector (ELSD), or any other evaporative detection method such as Charged Aerosol Detection (CAD) or Electrospray Ionization Mass Spectrometry (ESI-MS)

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