Carrageenan analysis. Part 3: Quantification in swine plasma

Area of Science:

• Food Science and Technology
• Analytical Chemistry
• Biochemistry

Background:

• Carrageenan is used to stabilize infant formulas.
• Detecting low molecular-weight carrageenan fractions (LMT) in biological samples is challenging due to strong protein interactions.
• A validated analytical method is required for safety studies.

Purpose of the Study:

• To develop and validate an analytical method for detecting the low molecular-weight tail (LMT) of carrageenan in swine plasma.
• To support a 28-day piglet feeding study using swine-adapted infant formulations stabilized with carrageenan.
• To assess the potential absorption of LMT carrageenan into the bloodstream.

Main Methods:

• Poligeenan, an acid-hydrolyzed carrageenan, was used as a surrogate analyte due to its weaker protein binding.
• Liquid chromatography-tandem mass spectrometry (LC-MS/MS) coupled with electrospray ionization (ESI) in negative-ion mode was employed.
• Method validation followed USFDA Good Laboratory Practice (GLP) Regulations.

Main Results:

• Poligeenan was successfully separated from plasma proteins using a salting procedure.
• A standard curve for poligeenan in swine plasma was established.
• The method achieved a lower level of quantification (LLOQ) of 10.0 µg/mL for poligeenan, with a quantification range of 10.0-100.0 µg/mL.

Conclusions:

• A robust and validated LC-MS/MS method was developed using poligeenan as a surrogate for carrageenan LMT.
• This method enables the detection of absorbed carrageenan LMT in swine plasma.
• The validated method supports the safety assessment of infant formulas containing carrageenan.