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Determination of l ‐hydroxyproline using hydrophilic interaction chromatography coupled to tandem mass spectrometry with lyophilized concentrated extraction in milk and dairy products
Author(s) -
Zhu Ailing,
Peng Tao,
Chen Dongdong,
Wang Ping,
Wang Guomin,
Wang Jinhua,
Jiang Haiyang,
Fan Chunlin,
Chen Ying
Publication year - 2014
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201400071
Subject(s) - chemistry , chromatography , extraction (chemistry) , mass spectrometry , tandem mass spectrometry
In this study, a screening and confirmation method for the determination of l ‐hydroxyproline (Hyp) as a target compound in milk and dairy products using high‐performance liquid chromatography with tandem mass spectrometry was developed. The samples were lyophilized after acidic hydrolysis, followed by cleanup with graphitized carbon black to remove pigments. Hyp was separated by a hydrophilic interaction chromatographic column and analyzed by high‐performance liquid chromatography with tandem mass spectrometry working with multiple reaction monitoring mode using an electrospray ionization interface in a positive‐ion mode. Average recoveries in spiked milk and dairy products ranged from 68.0 to 101.1% with relative standard deviations between 2.0 and 11.7% ( n = 7). A reagent‐matched standard calibration curve was used for quantification of Hyp, with linear correlation coefficient ( R 2 ) > 0.99 in the concentration range of 0.1–100 μg/mL. The LOQs were from 0.25 to 5 mg/kg, which were usually sufficient to verify the Hyp in samples. The confirmation concentration of Hyp ranged from 10 to 50 mg/kg.