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Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
41, Number 9, October 2003, pp. 460-466
HPLC Separation of Tetracycline Analogues: Comparison Study
of Laser-Based Polarimetric Detection with UV Detection
Karno Ng[1], and Sean W. Linder[2]
[1]Department of Chemistry & Biochemistry, California State University
at San Marcos, San Marcos, CA 92096 and
[2]Department of Chemistry & Biochemistry, University of Arkansas, Fayetteville,
AR 72701
A sensitive and specific high-performance liquid chromatography
(HPLC) method based upon laser-based polarimetric detection is developed for
the determination of six tetracycline analogues. By interfacing the laser-based
polarimeter online with an HPLC system, the specific rotation of each analogue
is obtained as compounds elute from the separation system. The six structurally
similar tetracycline analogues exhibit significant differences in specific
rotations. The experiments suggest that specific rotation can be useful in
identifying closely related tetracycline analogues. Linear relationships are
found to be in the range of 0.342–0.0043 mg for the tetracycline analogues.
Five of the six analogues exhibit excellent linearity (R2 value ≥ 0.99).
The polarimetric results are compared with UV detection. The HPLC–laser-based
polarimetric detection instrument is able to quantitate the studied tetracycline
analogues with high precision, accuracy, and sensitivity, which make it useful
for the development of a standard method for the determination of tetracyclines
in biological specimens. The performance of the HPLC–polarimetric system
for the analysis of tetracyclines in a biological matrix is evaluated. The
selectivity of polarimetric detection provides a distinct advantage in the
analysis of tetracycline analogues in milk. The HPLC–polarimetric system
provides a rapid and sensitive technique that involves minimal sample cleanup
and pretreatment for the analysis of tetracyclines in milk.
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