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Quantification of 5‐aminolevulinic acid by CE using dynamic p H junction technique
Author(s) -
Hsieh BoChuan,
Chen Richie L. C.,
Tsai Tsuimin
Publication year - 2013
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.201200889
Subject(s) - reproducibility , chromatography , chemistry , calibration curve , analytical chemistry (journal) , matrix (chemical analysis) , phosphate buffered saline , high performance liquid chromatography , resolution (logic) , detection limit , artificial intelligence , computer science
An online dynamic p H junction preconcentration method was developed for quantification of 5‐aminolevulinic acid (ALA) by CE with the separation time less than 6 min. The optimal dynamic p H junction of ALA was carried out between p H 9.3 borate buffer (BGE, 40 mM) and p H 2.5 phosphate buffer (sample matrix, 40 mM) when 4.1 cm of sample plug was hydrodynamically injected into an uncoated fused‐silica capillary (48.5 cm in length, id of 50 μm). If a 24 kV separation voltage was applied, the calibration curve of ALA peak area (200 nm) showed good linearity ( R 2 = 0.9991) ranging from 0.01 to 6.5 mg/mL. The reproducibility of this system was excellent with RSD s ( n = 10) of 2.5% for peak area response and 0.6% for migration time at ALA concentration of 0.5 mg/mL. The LOD was evaluated as 1.0 μg/mL ( S / N > 3). Compared to conventional CE procedure, the sensitivity was successfully improved over 50‐fold. The analytical results of pharmaceutical formulations show a good agreement with those by HPLC ( r = 0.94).