Development of a screening method for the determination of carotenoids in ham sausage, juice, and cookies by an improved Bligh-Dyer method and LC-MS/MS.

A screening method of 23 carotenoids in foods, such as ham sausage, juice, and cookies, was proposed using an improved Bligh-Dyer method that can satisfy the extraction requirements of common carotenoids with different physicochemical properties, including free and esterified carotenoids, by collecting the aqueous and organic phases simultaneously. Purification was then performed by loading the aqueous phase onto a hydrophile-lipophile balance (HLB) column; a methanol-water solution was used for washing, and the organic phases and additional chloroform were used for elution. By optimizing this step, interference from hydrophilic compounds and neutral triglycerides was effectively eliminated, and the matrix suppression effect after purification was greater than -16.3%. Finally, the extract was analyzed by ultrahigh performance liquid chromatography-quadrupole-orbitrap high-resolution mass spectrometry. The results showed that the 23 carotenoids showed a good linear relationship in the range of 0.01-0.2 µg/mL, and the limits of quantification (signal-to-noise ratio ≥10) were from 0.02 to 0.05 mg/kg. The average recoveries were 80.1%-98.7%, with relative standard deviation ≤10%. The proposed method can simultaneously identify and quantify 23 carotenoids in foods with high throughput, sensitivity, and reliability.