高通量高效液相色谱法快速检测西红花中15种非法添加色素

建立高通量的高效液相色谱法同时检测15种西红花中非法添加色素,对于阳性样品采用液相色谱–质谱联用法确证。采用Poroshell 120 EC–C18色谱柱(100 mm×4.6 mm,2.7 μm)分离,以甲醇–0.02 mol/L乙酸铵溶液(用氨水调pH至5.9±0.05)进行梯度洗脱,流量为1.2 mL/min,柱温为35 ℃,以450 nm和500 nm双波长检测。缓冲盐pH值和流动相流量对色谱条件影响大,需严格控制。该方法重现性好,各物质分离度均大于1.5,标准曲线的线性相关系数r为0.998~1.000,样品高、中、低浓度的加标回收率为70.1%~93.1%,测定结果的相对标准偏差为1.2%~6.6%(n=6),检出限为0.01~0.05 mg/kg。该方法精密度良好,分析时间短,分析结果稳定,可用于中药材西红花中15种非法添加色素的快速筛查和含量测定,同时能为相关部门制定监督抽检中药材染色掺伪的补充检验方法。

Rapid detection of 15 illegal additive pigments in Crocus Sativus by high throughput HPLC

A high throughput high performance liquid chromatography(HPLC)was established for the simultaneous determination of 15 pigments added illegally in Crocus Sativus and the positive samples were confirmed by liquid chromatography mass spectrometry(LC–MS).The chromatographic column Poroshell 120 EC–C18(100 mm×4.6 mm,2.7μm)was used as separation column,gradient elution was conducted using methanol–0.02 mol/L ammonium acetate solution(adjusted to a pH of 5.9±0.05).The column temperature was 35℃,and the flow rate was 1.2 mL/min,the detection wavelength was 450 nm and 500 nm.The chromatographic conditions were affected by the pH value and flow rate of buffer salts which should be controlled strictly.The method has good reproducibility,the separation degree of each substance was more than 1.5.The linear correlation coefficient(r)of the standard curve was 0.998–1.000,the standard recoveries of high,medium and low concentration addition were 70.1%–93.1%with the RSD of 1.2%–6.6%(n=6),the detection limit was 0.01–0.05 mg/kg.The method can be used for rapid screening and content determination of 15 pigments added illegally in Crocus Sativus with the good precision,short analysis time,stable analysis result,and it can also be used for the establishment of a supplementary inspection method to supervise and spot-check the adulteration and dyeing of crocus sativus.