Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume 45,
Number 4, April 2007, pp.195-199
Quantitation of Glucosamine From Shrimp Waste Using
HPLC
J. López-Cervantes, D.I. Sánchez-Machado,
and K.E. Delgado-Rosas
Departamento de Biotecnología y Ciencias Alimentarias,
Instituto Tecnológico de Sonora, P.O. Box 541, Cd. Obregón,
Sonora, México
This work presents a high-performance liquid chromatography
(HPLC) method for the quantitation of glucosamine in chitin. The
method includes an acid hydrolysis of chitin. The chromatographic
separation is achieved using a Hypersil ODS 5-µm column
(250 ¥ 4.6 mm) at 38°C, with precolumn derivatization
with 9-fluorenylmethyl-chloroformate and UV detection (l = 264
nm). The mobile phase is a mixture of mobile phase A [30mM ammonium
phosphate (pH 6.5) in 15:85 methanol–water (v/v)], mobile
phase B [15:85 methanol–water (v/v)], and mobile phase C
[90:10 acetonitrile–water (v/v)], with a flow rate of 1.2
mL/min. The HPLC method proposed showed adequate repeatability
(relative standard deviation, 5.8%), accuracy (92.7% recovery),
and sensitivity, with a detection limit of 2 µg/mL. The
method is successfully applied to the quantitation of glucosamine
for the determination of the purity of chitin from shrimp waste.
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