Single Laboratory Validation of a Method for Determination of Glucosamine in RawMaterials and Dietary Supplements Containing Glucosamine Sulfate and/or Glucosamine Hydrochloride by High-Performance Liquid Chromatography with FMOC-Su Derivatization

Abstract

Single laboratory validation of a method for determination of glucosamine in raw materials and dietary supplements containing glucosamine sulfate and/or glucosamine hydrochloride by with high-performance liquid chromatography FMOC-Su derivatization. Tests with 2 blank matrixes containing SAMe, vitamin C, citric acid, chondroitin sulfates, methylsulfonylmethane, lemon juice concentrate, and other potential interferents showed the method to be selective and specific. Eight calibration curves prepared over 7 working days indicated excellent reproducibility with the linear range at least over 2.0–150 μg/mL, and determination coefficients >0.9999. Average spike recovery from the blank matrix (n = 8 over 2 days) was 93.5, 99.4, and 100.4% at respective spike levels of 15, 100, and 150%, and from the sample matrix containing glucosamine (n = 3) was 99.9 and 102.8% at respective levels of 10 and 40%, with relative standard deviations <0.9%. The method was also applied to 12 various glucosamine finished products and raw materials. The stability tests confirmed that glucosamine–FMOC-Su derivative once formed is stable at room temperature for at least 5 days. Limit of quantitation was 1 μg/mL and limit of detection was 0.3 μg/mL. The method is ready to proceed for the collaborative study.

Enhancing the specificity of chitin determinations through glucosamine analysis via ultra-performance LC-MS

“…Although 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is suggested as derivatisation reagent for glucosamine in a previous study (36), the AQC derivative was also not retained sufficiently on the current system. Consequently, FMOC-Cl was used as derivatisation reagent as it is less polar compared to AQC and is also used in several studies on the chromatographic separation of glucosamine (19,21,37). Since FMOC-Cl reacts both with primary and secondary amines but not with amides, only the separation of glucosamine was optimised.…”

Kinetic Models for Glucosamine Production by Acid Hydrolysis of Chitin in Five Mushrooms

“…Glucosamine was determined by high-performance liquid chromatography (HPLC) according to the AOAC official method 2005.01 with minor modification [16,18,19]. Briefly, 0.75 μL trimethylamine was added to 1 mL sample solution and the mixture was allowed to react for at least 12 h at room temperature.…”

METODE HPLC-DAD UNTUK ANALISIS GLUKOSAMIN PADA TERIPANG KERING DENGAN DERIVATISASI PREKOLOM (HPLC-DAD Method for Glucosamine Analysis in Dried Sea Cucumber with Precolumn Derivatization)

“…In order to ensure that the products contain enriched glucosamine, an analytical method with sufficient accuracy, sensitivity and selectivity should be developed. Previous studies have reported the glucosamine content in shrimp (Lopez-Cervantes et al, 2007), chitin (Zhu et al, 2005), raw materials (Zhou, Waszkuc, & Mohammed, 2004) and human plasma (Huang et al, 2006;Ibrahim & Jamali, 2010;Song et al, 2012). Several papers have reviewed the nutritional and medicinal benefits of sea cucumber, including glucosamine (Bordbar et al, 2011;Khotimchenko, 2018;Pangestuti & Arifin, 2018).…”

“…A number of reagents such as 9fluorenylmethyl chloroformate (Fmoc-Cl) (Hami et al, 2013;Huang et al, 2006;Zhu et al, 2005), phenylisothiocyanate (PITC) (Du, White, & Eddington, 2004;Tekko, Bonner, & Williams, 2006), 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) (Wang et al, 2008), and N-(9fluorenyl-methoxycarbonyloxy) succinimide (Fmoc-Su) (Zhou et al, 2004) have been reportedly used as derivative reagents. Fmoc-Cl was used as a derivative reagent in this study, as many research have proved its effectiveness and stability to form glucosamine-Fmoc adduct, as illustrated in Figure 1 (Zhu et al, 2005).…”