超高效液相色谱-二极管阵列法测定清肺止咳散中非法添加的4种解热镇痛药

Determination of four kinds of antipyretic and analgesic drugs illegally added in Qingfeizhike powder by UPLC-PDA method

建立超高效液相色谱-二极管阵列检测法(UPLC-PDA法)测定清肺止咳散中非法添加对乙酰氨基酚、氨基比林、安替比林、安乃近4种药物。采用反相超高效液相色谱法,用带有二极管阵列检测器的高效液相色谱仪(UPLC-PAD)对对乙酰氨基酚、氨基比林、安替比林、安乃近4种解热镇痛药进行色谱分离和快速筛查。以ACQUITY UPLC HSS T3色谱柱为分离柱,柱温35℃;流动相体系为0.05 M醋酸铵水溶液(A项)、甲醇(B项),进行梯度洗脱;流速0.4 mL/min;进样量5μL;检测波长为245 nm。通过保留时间、光谱图、峰面积等参数对对乙酰氨基酚进行定性定量检测。结果显示,对乙酰氨基酚、氨基比林、安替比林在2~100μg/mL的范围内线性关系良好(R^2>0.999),安乃近在4~200μg/mL的范围内线性关系良好(R^2>0.999);添加对乙酰氨基酚、氨基比林、安替比林各2 mg/g,添加安乃近4 mg/g,信噪比(S/N)>3,确定为方法的检测限;添加对乙酰氨基酚、氨基比林、安替比林各5 mg/g,添加安乃近10 mg/g,信噪比(S/N)>10,确定为方法的定量限;清肺止咳散在3个添加水平的回收率为90%~105%,批内批间变异系数均小于10%,准确度和精密度良好,完全满足检测需求。该方法色谱分离较好,分析速度较快,前处理简单,适用于清肺止咳散中非法添加对乙酰氨基酚等4种解热镇痛药物的定性定量检测。

To establish an ultra-high performance liquid chromatography-diode array method(UPLC-PDA) for the determination of acetaminophen,aminopyrine,antipyrine and analgin in Qingfeizhike powder. Four antipyretic and analgesic drugs,acetaminophen,aminopyrine,antipyrine and analgin,were separated and screened by reversed phase high performance liquid chromatography(RP-HPLC) and high performance liquid chromatography(UPLC-PAD) with diode array detector. The ACQUITY UPLC HSS T3 column was used as the separation column,the column temperature was 35 ℃,the mobile phase system was 0.05 m ammonium acetate aqueous solution(A) and methanol(B),the flow rate was 0.4 mL·min-1,the injection volume was 5 μL,and the detection wavelength was 245 nm. The retention time,spectrum and peak area were used to detect acetaminophen qualitatively and quantitatively. The linear range of acetaminophen,aminopyrine and antipyrine was 2~100 μg·mL-1(R2>0.999),and the linear relationship was good in the range of 4~200 μg·mL-1(R2>0.999). The detection limit of the method was determined by adding 2 mg·g-1 of acetaminophen,aminopyrine and antipyrine,adding 4 mg·g-1 of analgin,signal to noise ratio(S/N)>3. The quantitative limit of the method was determined by adding 5 mg·g-1 of acetaminophen,aminopyrine and antipyrine,adding 10 mg·g-1 of analgin,signal-to-noise ratio(S/N)>10,the recovery of Qingfeizhike powder was 90% ≤105%,the coefficient of variation between batches was less than 10%,and the accuracy and precision were good. The results showed the method had the advantages of good chromatographic separation,fast analysis speed and simple pretreatment. It is suitable for the qualitative and quantitative detection of four antipyretic and analgesic drugs,such as the illegal addition of acetaminophen and other antipyretic and analgesic drugs in Qingfeizhike powder.