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題名 | Exploratory Study of Estimating Haze-Active Protein in Beverage Model System by Alizarin Red S=利用染劑Alizarin Red S定量在模擬飲料系統中霧狀活性蛋白質研究 |
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作者姓名(中文) | 楊景雍; | 書刊名 | 國立高雄海院學報 |
卷期 | 17 2002.12[民91.12] |
頁次 | 頁59-77 |
分類號 | 341.96 |
關鍵詞 | 霧狀活性蛋白質; 染劑吸附定量法; Alizarin red S; Haze-active protein; Dye binding; |
語文 | 英文(English) |
中文摘要 | 在飲料與釀造業最常被用來估算果汁或啤酒中霧狀活性蛋白質(haze-active protein, HAP)的方法為濁度法(nephelometric method)。但是侷於濁度(turbidmetric response)與HAP含量間有限的線性相關(linear relationships),在定量上的正確性有可議之處。如果能開發一種快簡的定量飲料中HAP的方法,有助於產品品質提升,例如在啤酒澄清穩定(clarification and stabilization)程序中,用此快簡的方法以便了解添加何種飲料澄清劑(或吸附劑)的效果最佳,以及澄清劑最佳用量。在模擬啤酒或果汁系統時,常添加霧狀活性蛋白質(HAP) gliadin與多酚類化合物(polyphenols)結合產生成霧狀顆粒的現象,以便探討啤酒或果汁中的HAP與多酚類化合物結合的行為。gliadin有純化的商業產品,故常當作定量飲料中HAP的標準蛋白。而定量HAP的目的是為了解飲料澄清穩定程度。 本研究發現一種結構近似多酚(polyphenols)的染劑分子Alizarin Red S對gliadin有較高的結合(binding)現象。Alizarin Red S被觀察到,其與飲料模擬系統中之gliadin在UV區的259nm產生最大吸光值變化,並可利用此吸光值變化定量溶液中HAP含量。此外ARS在模擬系統中與同濃度但不同霧狀活性程度的蛋白質(gliadin, BSA, papam, gelatin)進行結合試驗,這些蛋白質溶於模擬啤酒系統:pH4磷酸鹽溶液含5%的乙醇,這些蛋白質溶液與ARS在25℃下反應30分鐘。由實驗結果顯示ARS對gliadin有最大的吸光現象。此外在10mg/L~80mg/L範圍gliadin的濃度,與兩者結合產生吸光值變化間有良好線性相關(R2=0.96)。此染劑吸附定最法(dye binding method)簡單迅速,但應用量測飲料樣本時,為了避免飲料中小分子的干擾物影響定量結果,建議樣品先經分子量3500的透析膜進行透析後再行定量HAP。 |
英文摘要 | A new protein assay which involves the binding of Alizarin Red S (ARS) to haze-active protein (gliadin) in model beverage is studied. When the ARS dye reagent binds to haze-active protein (HAP), there is an absorbance change at 259 nm which can be monitored by difference spectra for estimation of the amount of haze-active protein ( HAP ) in samples. The current method designed to measure the protein fraction that actually forms haze in beverage (beer, juice etc.) is nephelometric procedure. However, it has the disadvantage common to other nephelometric methods of a severely limited linear range of response due to variations in the quantity and nature of the haze produced. It would be particularly useful to have a rapid and simple method for estimating the amount of haze-active protein in beverage, such as beer because this could be used to compare the effectiveness of various stabilization approaches, evaluate different lots of chillproofing materials, and estimate the extent of stabilization (and likely shelf-life) of a given beer. ARS was tested to bind protein gliadin (proline-rich, alcohol soluble protein) and other proteins ( BSA, papain, gelatin etc. ) in beer model system, pH4 buffer containing 5% ethanol with 30mtn incubation. The absorbance changes were recorded. ARS showed the strongest affinity to the model haze active protein gliadin. The multiple concentrations of gliadin gave a very good linear response to ARS (10 mg/L-80 mg/L and R square is 0.96). This assay is simply and rapid with the dye binding process virtually completed m approximately 30 min with good color stability for l hr. Because of some interfering substances in commercial beer, dialysis is suggested before this assay is conducted |
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