Question 1

What is the Meaning of C18 Column?

Accepted Answer

A C18 Column is a specific type of stationary phase utilized in High-Performance Liquid Chromatography (HPLC), an essential analytical technique in clinical laboratories for measuring hormones. The column packing material is silica chemically bonded with octadecyl (18-carbon) chains, creating a highly non-polar, hydrophobic surface. This configuration is critical for separating complex biological mixtures, enabling the precise isolation of various steroid hormones and their metabolites from a patient's sample matrix.

Question 2

What is the Origin of C18 Column?

Accepted Answer

This term originates from the field of analytical chemistry, specifically reversed-phase chromatography, which was developed to separate compounds based on their differential polarity. The designation 'C18' is a chemical shorthand for the 18-carbon alkane chain, octadecyl, covalently attached to the silica substrate. The adoption of this technology revolutionized clinical endocrinology by providing the necessary specificity to accurately measure multiple hormones simultaneously, overcoming the limitations of older immunoassay methods.

Question 3

What is the Mechanism of C18 Column?

Accepted Answer

The separation process is driven by the differential hydrophobic partitioning of the analytes between the non-polar C18 stationary phase and the polar mobile phase. Less polar compounds, such as many lipophilic steroid hormones, are retained longer due to stronger hydrophobic interactions with the C18 chains. Conversely, more polar metabolites pass through the column more quickly. This precise mechanism allows for the isolation of structurally similar hormonal compounds before their quantification by detectors like a mass spectrometer.

C18 Column

Meaning

Origin

Mechanism

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