鲍鱼内脏中天然牛磺酸的提取与检测

Extraction and determination of taurine from viscera of abalone(Haliotis discus hannai)

目的：以皱纹盘鲍内脏为原料,优化高纯度牛磺酸的提取工艺,并建立用高压液相色谱检测牛磺酸纯度的方法。方法经水煮、乙醇抽提、蒸发浓缩、沉淀、活性炭处理、结晶等步骤,获得高纯度天然牛磺酸。用高压液相色谱检测牛磺酸纯度的方法为：色谱柱：Discovery C18；流动相：甲醇-0.05 mol/L乙酸钠缓冲液(pH 5.3)(v/v,50：50)；流速：1 mL/min；检测波长：330 nm；柱温：室温。结果该提取工艺能从1 kg 鲍鱼内脏中提取得到天然牛磺酸3.07 g。采用该检测方法,牛磺酸的出峰时间为6.037 min,在1~20μg/mL浓度范围内线性关系良好(R2=0.9999),最低检出限为0.03μg/mL,最低定量限为0.12μg/mL,样品测定的平均回收率为99.44%(RSD=0.25%),方法的精密度和稳定性好(RSD<1%),用该方法检测提取得到的牛磺酸纯度为96.06%。采用红外光谱法(IR)对提取得到的鲍鱼内脏牛磺酸进一步作结构鉴定,发现它与标准品的红外特征吸收峰一致。结论获得了高纯度牛磺酸的提取工艺。本方法精密度和稳定性好,回收率高。

Objective The viscera from abalone (Haliotis discus hannai) was used as raw materials to isolate natural taurine and an HPLC method for the determination of the purity of taurine was established. Methods Natural taurine was isolated by combining methods of high temperature stewing, activated carbon treatment, ethanol extraction, concentration, precipitation and crystallization. The HPLC method for determination of the purity of taurine was performed on a Discovery C18 column eluted with a mobile phase of 50% methyl alcohol and 50% sodium acetate (pH 5.3) at flow rate of 1.0 mL/min at room tem-perature and detected by a UV detector at 330 nm. Results Taurine (3.07 g) was isolated from 1 kg ab-alone viscera by the extraction method. The retention time of taurine was at 6.037 min on HPLC and the method showed a good linearity within concentration of 1~20 μg/mL with R2 of 0.9999. The detection&nbsp;limit was 0.03 μg/mL and the quantitative detection limit was 0.12 μg/mL with an average recovery of sample of 99.44% (RSD=0.25%). The precision and stability of this method was optimal (RSD<1%). Us-ing this method, the purity of taurine extracted from abalone viscera reached 96.06%. The structure of the obtained taurine was further analyzed by infrared spectrometer and its characteristic absorption peak was consistent with that of the standard taurine. Conclusion The extraction method of taurine with high pur-ity was established. The determination method resulted in optimal precision and stability as well as a high recovery of sample.