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doi:10.1016/j.jchromb.2007.04.043 
Copyright © 2007 Elsevier B.V. All rights reserved.
Improved and simplified liquid chromatography/electrospray ionization mass spectrometry method for the analysis of underivatized glucosamine in human plasma
Received 15 January 2007;
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Abstract
Glucosamine is an amino monosaccharide reagent. It is difficult to assay using typical reversed-phase column due to the early elution, by optimizing the chromatographic conditions, especially the analytical column and the mobile phase composition, an improved analytical method was developed and validated, which offers rapid, sensitive and specific determination of glucosamine in human plasma. Following protein precipitation, the analyte and internal standard (valibose) were separated using an isocratic mobile phase on an Inertsil CN-3 column and detected by mass spectrometry in the multiple reaction monitoring mode using the respective precursor to product ion combinations of m/z 180/72 for glucosamine and m/z 252/198 for valibose. The chromatographic time was just 4.2 min for each sample, which made it possible to analyze more than 120 human plasma samples per day. The method exhibited a linear dynamic range of 4.00–4000 ng/mL for glucosamine in human plasma. The lower limit of quantification (LLOQ) was 4.00 ng/mL with a relative standard deviation of less than 10.9%. Acceptable precision and accuracy were obtained for the plasma concentrations over the standard curve range. By monitoring the two different MRM transitions, it was proved that no endogenous glucosamine was found in human plasma. The validated method has been successfully used to analyze human plasma samples for application in a bioequivalence study.
Keywords: Glucosamine; Chromatography optimization; LC/MS/MS; Plasma concentration
Article Outline
- 1. Introduction
- 2. Experimental
- 2.1. Chemicals
- 2.2. LC/MS/MS instruments and conditions
- 2.3. Preparation of calibration standards and quality control samples
- 2.4. Sample preparation
- 2.5. Bioanalytical method validation
- 2.6. Application to bioequivalence study
- 3. Results and discussion
- 3.1. Mass spectrometry conditions
- 3.2. Chromatographic conditions
- 3.3. Internal standard selection
- 3.4. Method validation
- 3.4.1. Selectivity and matrix effect
- 3.4.2. Linearity of calibration curves and lower limit of quantification
- 3.4.3. Precision and accuracy
- 3.4.4. Recovery and stability
- 3.5. Application
- 4. Conclusion
- References
