On-line sample cleanup and enrichment chromatographic technique for the determination of ambroxol in human serum |
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Authors: | Emara Samy Kamal Maha Abdel Kawi Mohamed |
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Institution: | Faculty of Pharmacy, Misr International University, Km 28 Ismailia Road, Cairo, Egypt. Emara_miu@yahoo.com |
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Abstract: | A sensitive and efficient on-line clean up and pre-concentration method has been developed using column-switching technique and protein-coated μ-Bondapak CN silica pre-column for quantification of ambroxol (AM) in human serum. The method is performed by direct injection of serum sample onto a protein-coated μ-Bondapak CN silica pre-column, where AM is pre-concentrated and retained, while proteins and very polar constituents are washed to waste using a phosphate buffer saline (pH 7.4). The retained analyte on the pre-column is directed onto a C(18) analytical column for separation, with a mobile phase consisting of a mixture of methanol and distilled deionized water (containing 1% triethylamine adjusted to pH 3.5 with ortho-phosphoric acid) in the ratio of 50:50 (v/v). Detection is performed at 254 nm. The calibration curve is linear over the concentration range of 12-120 ng/mL (r(2) = 0.9995). The recovery, selectivity, linearity, precision, and accuracy of the method are convenient for pharmacokinetic studies or routine assays. |
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