酶催化法合成(<i>R</i>)-1-(1-萘基)乙胺产品中蛋白残留的检测方法

吴俊; 李慧思; 张亚伟; 何魁芳; 朱坤丹; 谢升谷; 陶巧凤; 孙保国; 郑金琪

doi:10.13748/j.cnki.issn1007-7693.20240308

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

Method for Determination of Protein Residues in (R)-1-(1-naphthyl) Ethylamine Products Synthesized by Enzymatic Reaction

摘要

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)建立酶催化法合成(R)-1-(1-萘基)乙胺产品中蛋白残留的检测方法。
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=0.99)内线性关系良好；回收率>70%(n=3×6)；超滤离心后相当于收集了500 mg·mL⁻¹主成分溶液，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 To establish a method for the determination of protein residues in (R)-1-(1-naphthyl) ethylamine products prepared by enzyme-catalyzed reaction by ultrafiltration centrifugation combined with BCA method.
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 562 nm.
RESULTS The protein concentration exhibited a good linear relationship within the range of 25−500 μg·mL⁻¹(r²=0.99). The recovery rates were >70%(n=3×6). After ultrafiltration centrifugation, it was equivalent to collecting 500 mg·mL⁻¹ of the main component solution. The minimum detection limit of protein residues was 5 μg·mL⁻¹, 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.

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