UPLC-PDA测定鲜竹沥中4种抑菌剂的含量

Determination of Four Preservatives in Fresh Bamboo Juice by UPLC-PDA Method

目的:建立超高效液相色谱-二极管阵列检测器法(UPLC-PDA)检测市售鲜竹沥中苯甲酸、山梨酸、羟苯甲酯、羟苯乙酯4种常用抑菌剂的含量。方法:采用ACQUITYUPLCHSST3色谱柱(100mm×2.1mm,1.8µm),以乙腈(A)-0.02mol·L^(-1)乙酸铵溶液(B)为流动相梯度洗脱,流速0.3 mL·min^(-1),采用223 nm(苯甲酸)、254 nm(山梨酸、羟苯甲酯、羟苯乙酯)作为检测波长,进样量1µL,柱温30℃。结果:4种抑菌剂在其测定波长下均完全分离,4种抑菌剂平均加样回收率分别为97.9%、104.0%、99.2%、103.0%;重复性试验苯甲酸和羟苯乙酯峰面积的RSD(n=6)均小于1%;4种抑菌剂质量浓度与峰面积均呈良好线性关系(r均大于0.99945);且样品的稳定性较好,48 h内苯甲酸峰面积的RSD为1.47%,羟苯乙酯峰面积的RSD为0.2%,对13批鲜竹沥中的抑菌剂进行了检测,9批鲜竹沥检出苯甲酸(质量分数为0.07%~0.29%),2批鲜竹沥检出山梨酸(质量分数为0.21%~0.24%),2批鲜竹沥检出苯甲酸(质量分数为0.21%~0.22%)和羟苯乙酯(质量分数为0.025%~0.027%),均符合《中华人民共和国药典》2020版(四部)通则0181合剂项下要求。结论:建立了基于UPLC-PDA测定鲜竹沥中的4种常用抑菌剂含量的方法,该方法专属性、重复性、准确度良好,为鲜竹沥这一冷背品种的质量控制提供了参考。

To establish a ultrahigh performance liquid chromatography-diode array detector(UPLC-PDA)method to detect the contents of four commonly used preservatives in commercially available fresh bamboo juice,including benzoic acid,sorbic acid,methylparaben,and ethylparaben.Methods:The ACQUITY UPLC HSS T3 column(100 mm×2.1 mm,1.8µm)was used with a gradient elution of acetonitrile as mobile phase A and 0.02 mol·L^(-1) ammonium acetate as mobile phase B at a flow rate of 0.3 mL·min^(-1).The 223 nm(benzoic acid)and 254 nm(sorbic acid,methylparaben and ethylparaben)were used as detection wavelengths,the injection volume was 1µL,and the column temperature was 30℃.Results:The four preservatives were all completely separated at their determination wavelengths,and the average recoveries of the four preservatives were 97.9%,104.0%,99.2%and 103.0%,respectively;the RSDs(n=6)of the peak areas of benzoic acid and ethylparaben in the repeatability test were both<1%;the mass concentrations of the four preservatives showed a good linear relationship with the peak area(r all>0.99945);and the samples had a good stability.The RSD of the peak area of benzoic acid within 48 h was 1.47%,and the RSD of the peak area of ethylparaben within 48 h was 0.2%.The preservatives in 13 batches of fresh bamboo juice were detected,benzoic acid was detected in 9 batches of fresh bamboo juice(mass fraction of 0.07%-0.29%),sorbic acid was detected in 2 batches of fresh bamboo juice(mass fraction of 0.21%-0.24%),benzoic acid(mass fraction of 0.21%-0.22%)and ethylparaben(mass fraction of 0.025%-0.027%)were detected in 2 batches of fresh bamboo juice,all of which met the requirements in Section of Mixture of the General Rule 0181 in the Chinese Pharmacopoeia(2020 version)VolumeⅣ.Conclusion:A method for determining the contents of four commonly used preservatives in fresh bamboo juice is established based on UPLC-PDA,and the method has good specificity,reproducibility and accuracy,which provides a reference for the quality control of fresh bamboo juice,a product which is not used on a large scale.