酶催化法合成(R)-1-(1-萘基)乙胺产品中蛋白残留的检测方法研究

Study on the method for determination of protein residues in (R)-1-(1-naphthyl) ethylamine products synthesized by Enzymatic Reaction

关键词:  蛋白残留测定  BCA法  (R)-1-(1-萘基)乙胺  酶催化  超滤

Key words:  protein residue  BCA method  ultrafiltration  (R)-1-(1-naphthyl) ethylamine  enzyme catalysis

摘要:

目的 利用超滤离心结合2,2‘-联喹啉-4,4‘-二羧酸法(BCA法)建立酶催化反应制备的(R)-1-(1-萘基)乙胺产品中蛋白残留的检测方法研究。方法 采用Amicon? Ultra-15型超滤管对其溶液进行净化并对蛋白进行浓缩富集，结合2,2‘-联喹啉-4,4‘-二羧酸法(BCA法)测定 (R)-1-(1-萘基)乙胺中的蛋白残留含量，检测波长562 nm。结果 蛋白浓度在25~500 μg·mL-1（r2=0.995）内线性关系良好；回收率＞70%（n=4*3）；蛋白质残留的最低检出限为5 μg·mL-1，相当于主成分含量的0.001%。结论 本方法易操作、重复性好、准确度高，适用于酶催化反应制备的(R)-1-(1-萘基)乙胺产品中残留蛋白的测定。

Abstract:

OBJECTIVE By ultrafiltration centrifugation combined with BCA method, to establish a method for the determination of protein residues in (R)-1-(1- naphthyl) ethylamine products prepared by enzyme-catalyzed reaction. METHODS The Amicon? Ultra-15 ultrafiltration tube was used to purify the solution and enrich the protein. The protein residue in (R)-1-(1-naphthol) ethylamine was determined by BCA method with the detection wavelength of 562nm. RESULTS The protein concentration was 25-500 μg/mL (r2=0.995).The recovery rate was more than 70% (n=4*2*3).The minimum detection limit of protein residues was 5 μg/mL, which was equivalent to 0.001% of the principal component content. CONCLUSION This method is easy to operation, has good repeatability and high accuracy, and is suitable for the determination of residual protein in (R)-1-(1-naphthalene) ethylamine products prepared by enzymatic reaction.