A simple, rapid and specific method using high performance liquid chromatography with fluorescence detector (HPLC-FLD) was developed for quantification of glucosamine in supplements after derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). The chromatographic separation was achieved on a C18 column (150 mm × 4.6 mm × 5 µm) using a gradient mobile phase containing acetonitrile and water at a flow rate of 1.0 mL/min. The FLD detector was operated at excitation and emission wavelengths of 265 and 315 nm, respectively. The analytical method was validated for specificity, linearity, accuracy, and precision. The detector response for glucosamine was linear over the selected concentration range from 6.7 to 135 ppm with a correlation coefficient of 0.9988. The repeatability (RSD, n = 6) and reproducibility (RSD, n = 4) were from 2.54 to 3.23 % and from 2.24 to 3.07 %, respectively. The recovery was between 98.0 and 101.5 %. In addition, we also joined the interlaboratory proficiency testing program (code: H21.11) organized by the National Institute of Food Control with good result (z-score value was - 0.89 < 2). The method was successfully applied for the analysis of glucosamine in 15 supplement samples.
Glucosamine, HPLC-FLD, supplements.
[1]. J. W. Anderson, R. J. Nicolosi, and J. F. Borzelleca, “Glucosamine effects in humans: a review of effects on glucose metabolism, side effects, safety considerations and efficacy,” Food and Chemical Toxicology, vol. 43, no. 2, pp. 187-201, 2005.
[2]. J. Y. Reginster, R. Deroisy, L. C. Rovati, R. L. Lee, E. Lejeune, O. Bruyere, G. Giacovelli, Y. Henrotin, J. E. Dacre and C. Gossett, “Long-term effects of glucosamine sulphate on osteoarthritis progression: a randomised, placebo-controlled clinical trial,” The Lancet, vol. 357, no. 9252, pp. 251-256, 2001.
[3]. R. Muniyappa, R. J. Karne, G. Hall, S. K. Crandon, J. A. Bronstein, M. R. Ver, G. L. Hortin and M. J. Quon, “Oral Glucosamine for 6 Weeks at Standard Doses Does Not Cause or Worsen Insulin Resistance or Endothelial Dysfunction in Lean or Obese Subjects,” Diabetes, vol. 55, no. 11, pp. 3142-3150, 2006.
[4]. K. Vongnam, C. Muangnoi, P. Rojsitthisak, M. Sukwattanasinitt and P. Rashatasakhon, “A highly selective turn-on fluorescent sensor for glucosamine from amidoquinoline-napthalimide dyads,” Biosensors and Bioelectronics, vol. 86, pp. 472-476, 2016.
[5]. E. Václavíková and F. Kvasnička, “Isotachophoretic determination of glucosamine and chondroitin sulphate in dietary supplements,” Czech Journal of Food Sciences, vol. 31, pp. 55-65, 2013.
[6]. P. Chaisuwan, T. Kongprasertsak, A. Sangcakul, N. W. Smith, D. Nachapricha, P. Wilairat and K. Uraisin, “Direct injection of human serum and pharmaceutical formulations for glucosamine determination by CE-C4D method,” Journal of Chromatography B, vol. 879, no. 23, pp. 2185-2188, 2011.
[7]. S. Akamatsu and T. Mitsuhashi, “Development of a simple capillary electrophoretic determination of glucosamine in nutritional supplements using in-capillary derivatisation with o-phthalaldehyde,” Food Chemistry, vol. 130, no. 4, pp. 1137-1141, 2012.
[8]. J. Z. Zhou, T. Waszkuc and F. Mohammed, “Determination of glucosamine in raw materials and dietary supplements containing glucosamine sulfate and/or glucosamine hydrochloride by high-performance liquid chromatography with FMOC-Su derivatization: collaborative study,” Journal of AOAC International, vol. 88, no. 4, pp. 1048-58, 2005.
[9]. AOAC Official Method 2005.01, Glucosamine in Raw Materials and Dietary Supplements Containing Glucosamine Sulfate and/or Glucosamine Hydrochloride. High-Performance Liquid Chromatography with FMOC-Su Derivatization. First Action, 2005.
[10]. Y. Shao, R. Alluri, M. Mummert, U. Koetter and S. Lech, “A stability-indicating HPLC method for the determination of glucosamine in pharmaceutical formulations,” Journal of Pharmaceutical and Biomedical Analysis, vol. 35, no. 3, pp. 625-631, 2004.
[11]. Nguyễn Trung Dũng, Lê Thị Dung, Vũ Thị Kim Oanh, Vũ Thị Trang, “Xác định Glucosamin trong thực phẩm chức năng bằng phương pháp sắc ký lỏng hiệu năng cao kết hợp với detector tán xạ bay hơi (HPLC-ELDS),” Tạp chí Khoa học và Công nghệ, số 52, tr. 87-93, 2014.
[12]. Xiaolan Zhu, “Determination of Glucosamine in impure chitin samples by high-performance liquid chromatography” Carbohydrate Research, vol. 340, pp. 1732-1738, 2005.
[13]. AOAC, “Appendix F: Guidelines for standard method performance requirements,” AOAC Official Methods of Analysis, 9, 2012.