高效液相色谱法测定略阳乌鸡肌肉中的肌苷酸

以略阳乌鸡为研究对象,通过对色谱条件进行优化,建立了略阳乌鸡肌肉中肌苷酸的高效检测方法,并重点就不同前处理方法对略阳乌鸡肌肉中肌苷酸含量的影响进行研究。对样品进行冷冻干燥、脱脂和冷冻干燥脱脂处理,以0.05%磷酸三乙胺为提取溶剂提取肌肉中的肌苷酸,采用COSMOSIL Packed Column C₁₈(4.6 mm×250 mm,5μm)色谱柱,甲醇-磷酸三乙胺(体积比4∶96)为流动相进行梯度洗脱。结果显示,该方法下肌苷酸呈现良好的线性关系(R²>0.999),线性范围在12.2~97.6μg/mL;方法检测限分别为0.02、0.08μg/mL;方法精密度相对标准偏差为0.30%~4.78%;平均回收率为99%,相对标准偏差为2.71%。鲜肉样、冷冻干燥样、鲜肉去脂样、冷冻干燥去脂样的肌苷酸含量分别为0.014、0.018、0.023、0.029 g/kg。该实验建立了精确度高、重现性好的色谱方法;前处理方法采用先冷冻干燥再脱脂处理,肌苷酸的提取率高,为高效液相色谱法测定肌肉中肌苷酸含量的样品制备提供了思路。

Determination of inosinic acid in muscle of Lueyang black-bone chicken by HPLC

Lueyang black-bone chicken was used as the raw material in this study to establish an efficient detection method for inosinic acid in muscle of Lueyang black-bone chicken through optimizing chromatographic conditions.And the effects of different pretreatment methods on the content of inosinic acid in the muscle of Lueyang black-bone chicken was also be detected.Samples were subjected to freeze-drying,degreasing and freeze-drying degreasing.Meanwhile,inosinic acid in muscle was extracted with 0.05%triethylamine phosphate.A cosmosil packed column C 18(4.6 mm×250 mm,φ5μm)was used for gradient elution with methanol-triethylamine phosphate(4∶96)as mobile phase.By using this method,inosinic acid showed a good linear relationship(R ²>0.999)which range 12.2-97.6μg/mL.The detection limits of this method were 0.02μg/mL and 0.08μg/mL,respectively.Moreover,the precision RSD of the method was 0.30%-4.78%and the average recovery rate and RSD was 99%and 2.71%respectively.The contents of inosinic acid in fresh meat samples,freeze-dried samples,fresh meat defatted samples and freeze-dried defatted samples were 0.014,0.018,0.023 and 0.029 g/kg,respectively.In this experiment,a chromatographic method with high accuracy and good reproducibility was established.After freeze-drying and degreasing,the extraction rate of inosinic acid is high,which provides an idea for preparing samples for determination of inosinic acid content in muscle by HPLC.