摘要
微生物发酵法已成为生产1-脱氧野尻霉素(1-DNJ)的主流方法,但是发酵液基质较为复杂,检测其中1-DNJ较为困难且未见相关报道,因此进行了提示研究。取0.1 000g牦牛奶提取菌发酵液样品,用水定容至10 mL,超声溶解5 min后经0.22μm滤膜和活化好的RP前处理小柱净化,弃去前3 mL流出液,收集后续流出液2 mL进行离子色谱分析。采用Dionex CarboPacTMMA1阴离子分析柱,在柱温30℃、流量0.30 mL·min^(-1)条件下,以由1 000 mmol·L^(-1)氢氧化钠溶液和水组成的流动相进行梯度洗脱,并基于四电位波形脉冲安培检测器对1-DNJ进行定量检测。结果显示,1-DNJ的质量浓度在0.01~1.64 mg·L^(-1)内和对应的峰面积呈线性关系,检出限(3S/N)为0.63μg·L^(-1);对实际样品进行精密度和加标回收试验,测定值的相对标准偏差(n=6)为1.8%,回收率为98.8%~101%;方法用于5批次实际样品的分析,1-DNJ的最高检出量为1.34μg·g^(-1)。
Given that microbial fermentation has become the mainstream method for producing 1-deoxynojirimycin(1-DNJ),the matrix of the fermentation broth is relatively complex,and the detection of 1-DNJ is difficult with no relevant reports available.Therefore,the study mentioned by the title was conducted.The 0.1000 g of fermentation broth sample of bacteria extracted from yak milk was taken,its volume was made up to 10 mL with water,and the mixture was purified through a 0.22μm filter membrane and an activated RP pre-processing cartridge after ultrasonic dissolution for 5 min.The first 3 mL of eluate was discarded,and 2 mL of the subsequent eluate was collected for ion chromatography analysis.Using a Dionex CarboPacTM MA1 anion analytical column at column temperature of 30℃and flow rate of 0.30 mL·min^(-1),gradient elution was performed with the mobile phase consisting of 1000 mmol·L^(-1) sodium hydroxide solution and water.Quantification of 1-DNJ was carried out using a quadruple-potential waveform pulsed amperometric detector.It was shown that linear relationship between the mass concentrations of 1-DNJ and corresponding peak areas was kept in the range of 0.01–1.64 mg·L^(-1),with detection limit(3S/N)of 0.63μg·L^(-1).Tests for precision and spike recovery were performed on actual samples,showing RSDs(n=6)of the determined values of 1.8%,and recoveries in the range of 98.8%–101%.The proposed method was applied to the analysis of 5 batches of actual samples,and the highest detected amount of 1-DNJ was 1.34μg·g^(-1).
作者
杨若璇
侯畅
张丹丹
王晓花
吴正钧
赵思嘉
栾绍嵘
YANG Ruoxuan;HOU Chang;ZHANG Dandan;WANG Xiaohua;WU Zhengjun;ZHAO Sijia;LUAN Shaorong(School of Chemistry and Molecular Engineering,East China University of Science and Technology,Shanghai 200237,China;Dairy Research Institute,State Key Laboratory of Dairy Biotechnology,Bright Dairy&Food Co.,Ltd.,Shanghai 200436,China)
出处
《理化检验(化学分册)》
北大核心
2025年第11期1341-1346,共6页
Physical Testing and Chemical Analysis(Part B:Chemical Analysis)
关键词
离子色谱法
安培检测器
1-脱氧野尻霉素
牦牛奶提取菌发酵液
ion chromatography
amperometric detector
1-deoxynojirimycin
fermentation broth of bacteria extracted from yak milk
