UPLC-MS/MS法快速测定中药及保健食品中非法添加17种抗炎镇痛类化学药的研究

Rapid determination of 17 anti-inflammatory and analgesic chemicals illegally added into Chinese patent medicine and health foods by UPLC-MS/MS

目的建立一种快速、准确检测中药及保健食品中非法添加17种抗炎镇痛类化学药（对乙酰氨基酚、阿司匹林、非那西丁、马来酸氯苯那敏、罗非昔布、吡罗昔康、氯诺昔康、美洛昔康、醋酸泼尼松、舒林酸、萘普生、醋酸地塞米松、保泰松、奥沙普秦、塞米昔布、双氯芬酸钠、吲哚美辛）的方法。方法采用UPLC-MS/MS法,以Waters Acquity BEH-C18柱（100 mm×2.1 mm,1.7μm）为色谱柱,以0.1%甲酸甲醇溶液（A）-0.1%甲酸水溶液（B）为流动相,梯度洗脱：0-4 min,40%A;4-5 min,40%-50%A;5-6 min,50%-60%A;6-12 min,60%-80%A;12-13 min,80%A;13-14 min,80%-40%A;体积流量0.2 mL/min,柱温40℃。选择ESI离子源、多反应监测（MRM）模式测定17种临床常用的抗炎镇痛类化学药,通过比较MRM通道中样品峰与对照品峰的分子离子峰、二级碎片离子峰、色谱保留时间等信息确定添加的化学药物,并根据外标法以质谱峰面积计算添加药物的准确量。结果在上述色谱及质谱条件下,对乙酰氨基酚、阿司匹林、非那西丁、马来酸氯苯那敏、罗非昔布、吡罗昔康、氯诺昔康、美洛昔康、醋酸泼尼松、舒林酸、萘普生、醋酸地塞米松、保泰松、奥沙普秦、塞米昔布、双氯芬酸钠、吲哚美辛17种化学药物的分离度良好,方法检测限（LOD）均在0.3-5.0 ng/g,定量限（LOQ）均在0.9-15.0 ng/g,加样回收率均在90.5%-113.8%。样品中检出了对乙酰氨基酚、醋酸泼尼松、双氯芬酸钠、吲哚美辛、马来酸氯苯那敏、萘普生。结论方法简便、准确,灵敏度高,可作为抗炎镇痛类中药及保健食品中非法添加化学药的定性定量测定方法。

Objective To estabilsh a rapid and accurate method for the determination of 17 anti-inflammatory and analgesic chemicals（paracetamol, aspirin, phenacetin, chlorpheniramine maleate, rofecoxib, piroxicam, lornoxicam, meloxicam, prednisone acetate, sulindac, naproxen, dexamethasone acetate, phenylbutazone, oxaprozin, celecoxib, diclofenac sodium, and indomethacin） which were illegally added into the Chinese patent medicines（CPM） and the health foods. Methods The UPLC-MS/MS method was adopted. The samples were extracted with methanol by ultrasonic processing and separated on a Waters Acquity BEH C18 column（100 mm × 2.1 mm, 1.7 μm） with 0.1% formic acid methanol（A） and 0.1% formic acid water（B） as the mobile phase by gradient elution（0—4 min, 40% A; 4—5 min, 40%—50% A; 5—6 min, 50%—60% A; 6—12 min, 60%—80% A; 12—13 min, 80% A; 13—14 min, 80%—40% A） at a flow rate of 0.2 mL/min, and the column temperature was 40 ℃. A positive-ion（ESI＋） source and a MRM mode were used to separately and quantitatively determinate the 17 anti-inflammation and analgesia chemicals. The obtained molecular ions, fragment ions, and retention time for MRM channels were used to identify the 17 chemicals by comparison with those of reference substances. The obtained peak areas were used to determinate the accurate content of chemicals in commonly used drugs in clinic. Results A good resolution of the 17 chemicals, including paracetamol, aspirin, phenacetin, chlorpheniramine maleate, rofecoxib, piroxicam, lornoxicam, meloxicam, prednisone acetate, sulindac, naproxen, dexamethasone acetate, phenylbutazone, oxaprozin, celecoxib, diclofenac sodium, and indomethacin was obtained under this UPLC and MS/MS condition. The limits of qualitation and quantitation were in the range of 0.3—5.0 and 0.9—15.0 ng/g. The standard addition recoveries were in the range of 90.5%—113.8%. The six components in samples（paracetamol, prednisone acetate, diclofenac sodium, indometacin, chlorpheniramine maleate, and naproxen） were detected. Conclusion The method is simple, accurate, and has high sensitivity, which can be used for the qualitative and quantitative determination of illegally added chemicals in CPM and health foods.