title: Carrageenan analysis. Part 3: Quantification in swine plasma.
abstract: Development and validation of this method was conducted to support a 28-day piglet feeding study of swine-adapted infant formulations stabilised with carrageenan. The validation was performed in accordance with USFDA Good Laboratory Practice (GLP) Regulations and associated current bioanalytical guidelines. Separation of carrageenan from plasma protein was unsuccessful using saturated sodium chloride due to the extremely strong cross-linking interactions between carrageenan and protein. Poligeenan is the deliberately acid-hydrolysed low molecular weight polygalactan non-food product produced from carrageenan. Poligeenan molecules are nearly identical to carrageenan molecules with respect to molecular structure, the primary difference being molecular weight. These poligeenan molecules have similar molecular weight when compared with the lowest molecular weight fraction of carrageenan called the low molecular-weight tail (LMT). Poligeenan was separated from plasma protein using the salting procedure, this being due to the significantly weaker interaction with protein caused by its shorter molecular chain length. Thus, poligeenan was applied as a chemical analyte surrogate for the LMT of carrageenan solely for the development and validation of the method. This method was used to try to detect the LMT of the carrageenan test material during the 28-day piglet feeding study, and if such was absorbed into the bloodstream. Successful development and validation of the method was achieved using LC-MS/MS coupled with ESI in negative-ion mode. A standard curve of instrument response versus poligeenan concentration was developed using swine plasma spiked with a range of poligeenan concentrations. The lower level of quantification (LLOQ) of poligeenan was 10.0 µg ml⁻¹, and the quantification range was 10.0-100.0 µg ml⁻¹. No animals were fed poligeenan.
authors.affiliation: a Celtic Colloids Inc ., Topsham , ME , USA.
authors.forename: William R
authors.initials: WR
authors.lastname: Blakemore
authors.forename: Ashley F
authors.initials: AF
authors.lastname: Brant
authors.forename: Jonathan G
authors.initials: JG
authors.lastname: Bissland
authors.forename: Natalie D
authors.initials: ND
authors.lastname: Bissland
mesh_terms: D000821:Animal Feed; D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers / Q000097:blood / Q000737:chemistry; D002351:Carrageenan / Q000097:blood* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D005520:Food Preservatives / Q000032:analysis* / Q000737:chemistry; D006358:Hot Temperature; D006801:Humans; D006868:Hydrolysis; D041943:Infant Formula / Q000737:chemistry; D007231:Infant, Newborn; D057230:Limit of Detection; D008970:Molecular Weight; D009994:Osmolar Concentration; D011134:Polysaccharides / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D034421:Sus scrofa; D053719:Tandem Mass Spectrometry
chemical_list: D015415:Biomarkers; D005520:Food Preservatives; D011134:Polysaccharides; C014157:poligeenan; D002351:Carrageenan
keywords: ESI; LC-MS/MS; carrageenan; poligeenan; swine plasma