膠囊錠狀食品中葡萄糖胺之檢驗方法探討   全文下載

Development of an Analytical Method for Glucosamine Quantification in Foods in Capsule and Tablet Form

葡萄糖胺（Glucosamine），係由蝦蟹甲殼萃取，經乾燥、去蛋白質、礦物質處理後而生成。前行政院衛生署於民國94年公告含葡萄糖胺製品之管理規定，其中葡萄糖胺及葡萄糖胺鹽酸鹽（Glucosamine hydrochloride）兩種成分列為食品管理。一般常見葡萄糖胺食品以膠錠、飲品及軟膠囊形式存在。衛生福利部食品藥物管理署（下稱食藥署）於106年公開之建議方法「食品中游離胺基酸、葡萄糖胺及牛磺酸之檢驗方法」不適用於軟膠囊檢體，且未將葡萄糖胺2種首旋異構物均納入含量計算。本研究係將非油性及油性基質檢體經不同前處理流程，前者以去離子水萃取均質樣品，後者先以乙酸乙酯溶解樣品，再以去離子水重複萃取兩次，收集下層液，以鄰苯二甲醛溶液進行衍生化反應。檢液以Poroshell HPH-C18（2.7μm, 3.0 mm×10 cm）管柱及超高效液相層析儀搭配光二極體陣列檢出器（UPLC-PDA），以磷酸二氫鈉溶液（pH 7.8）與乙腈／甲醇／水（4.5:4.5:1, v/v/v）溶液，90:10（v/v）比例作為移動相，以流速1 mL/min進行沖提，可於5分鐘內良好分離葡萄糖胺之2種首旋異構物。於粉狀膠錠及油性軟膠囊空白樣品中分別添加相當於5、25及50 mg/g葡萄糖胺之葡萄糖胺鹽酸鹽標準品進行確效試驗，其平均回收率分別為97-110%及98-105%，變異係數為2.4-3.6%及1.4-7.9%，皆符合「食品化學檢驗方法之確效規範」。以此方法進行7件市售產品中葡萄糖胺之含量檢測，檢測值介於21.6-372.7 mg/g，均高於標示值80%，符合「包裝食品營養標示應遵行事項」之含量標示規定。本研究優化層析條件及前處理流程，提供更精確可靠之檢測方法，適用於廠商自主管理及後市場監測。

Glucosamine extracted from shrimp and crab shell, after drying, de-protein and de-mineral treatment, is often made into the form of capsule, beverages and soft capsule in commercial food products. Two peaks of glucosamine can be found in the HPLC profile, due to the interconversion of the natural stereoisomers (α and β) in aqueous solution. “Method of Test for Free Amino Acids, Glucosamine and Taurine in Foods”, pronounced in 2017 by the Taiwan Food and Drug Administration, is not applicable to quantify glucosamine in soft capsule, besides, only one of the isomers was used for calculation. In this study, the non-oily and oily samples were analyzed using different extraction treatment. The former was extracted with deionized water, and the latter was dissolved with ethyl acetate before extracted with deionized water. The filtrate collected from the aqueous layer was derived with o-phthalaldehyde solution, and then analyzed by an ultra-high performance liquid chromatography system with a Poroshell HPH-C18 column (2.7 μm, 3.0 mm × 10 cm) and a photodiode array detector. The derivatized samples were subjected to 1 mL/min isocratic elution with sodium dihydrogen phosphate solution (pH 7.8) and acetonitrile/methanol/water (4.5:4.5:1, v/v/v) solution at the ratio of 90:10 (v/v). Glucosamine can be detected within 5 minutes, and the content is calculated by the sum of both stereoisomers. The average recoveries of glucosamine in the non-oily and oily samples spiked at the levels of 5, 25 and 50 mg/g ranged between 97-110% and 98-105%, and the coefficients of variation were 2.4-3.6% and 1.4-7.9%, respectively. All results showed that the method had good precision and accuracy. Seven commercial products were investigated, the results showed that the contents of glucosamine in the samples were in the range of 21.6-372.7 mg/g. The detected values were all higher than 80% of the labeled value, which were in compliance with the regulation.