高效液相色谱检测发酵液中乙酰辅酶A  

Detection of acetyl-CoA in fermentation broth by high-performanceliquid chromatography

作　　者：唐艳 高鹏 吴涛 TANG Yan;GAO Peng;WU Tao(Tongliao Meihua Bio-tech Co.,Ltd.,Tongliao 028024,China;Meihua Biotech(Langfang)Co.,Ltd.,Langfang 065001,China)

机构地区：[1]通辽梅花生物科技有限公司,内蒙古通辽028024 [2]廊坊梅花生物技术开发有限公司,河北廊坊065001

出　　处：《发酵科技通讯》2023年第4期187-190,共4页Bulletin of Fermentation Science and Technology

基　　金：河北省氨基酸工程技术研究中心创新平台建设资助项目(131000021)。

摘　　要：在发酵代谢过程中,乙酰辅酶A起着重要作用,为研究其在发酵液中的残留情况,建立了一种快速检测发酵液中乙酰辅酶A残留的液相检测方法。以安捷伦Polaris 5 C18-A(250 mm×4.6 mm,5μm)为色谱柱,流动相A为40 mmol/L磷酸钠水溶液,流动相B为V(40 mmol/L磷酸钠溶液)∶V(乙腈)=800∶200的混合均匀的溶液,梯度洗脱,流速为1 mL/min,紫外检测波长为254 nm。研究结果表明:乙酰辅酶A的出峰时间为18 min,线性回归系数R^(2)=1,加标回收率为99.5%,相对标准偏差(n=6)为0.5%,重现性好,以信噪比S/N为3计,检出限为0.5 mg/L。将该方法应用于发酵液中乙酰辅酶A的测定,具有简便高效、结果准确,以及重现性好的优势。Acetyl-CoA plays a crucial role in the process of fermentation metabolism.In order to accurately determine the residual amount of acetyl-CoA in fermentation fluid,a liquid phase detection method was established,allowing for rapid detection.The method employed an Agilent Polaris 5 C18-A(250 mm×4.6 mm,5μm)column with mobile phase A consisting of a 40 mmol/L sodium phosphate solution,and mobile phase B consisting of an evenly mixed solution of V(40 mmol/L sodium phosphate solution)∶V(acetonitrile)=800∶200 with gradient elution.The flow rate was set at 1 mL/min,and the wavelength of the UV detector was set to 254 nm.The results of the experiment demonstrated that the peak time of acetyl-CoA was 18 minutes,with a linear regression coefficient(R^(2))of 1,a standard recovery rate of 99.5%,a relative standard deviation(n=6)of 0.5%,and excellent reproducibility.Furthermore,the detection limit was determined to be 0.5 mg/L with a signal to noise ratio(SNR)of 3.In summary,this liquid phase detection method is both simple and efficient,providing accurate and reproducible results for the determination of acetyl-CoA in fermentation broth.

关 键 词：发酵液 乙酰辅酶A 液相 检测 

分 类 号：Q815[生物学—生物工程]