7 8 Vol. 7 No. 8 2016 8 Journal of Food Safety and Quality Aug., 2016 9 孙 亮, 张 蓉, 邬 国 庆, 邸 铮, 刘 齐, 张 华 珺 * (, 102206) 摘 要 : 目 的 9 方 法, Inertsil ODS-3, 0.02 mol/l - ; 1.0 ml/min; 230 nm, 30,, 9 结 果 20 min 9 9 1.0~100 μg/ml, 0.999;, 77.9%~102.9%, 0.42%~5.8% 结 论,,, 9 关 键 词 : ; ; Simultaneous determination of 9 kinds of lipid lowering drugs illegally added in health food by high performance liquid chromatography SUN Liang, ZHANG Rong, WU Guo-Qing, DI Zheng, LIU Qi, ZHANG Hua-Jun * (Beijing Institute for Drug Control, Beijing 102206, China) ABSTRACT: Objective To establish a method for the simultaneous determination of 9 kinds of drugs (nicotinic acid, pravastatin sodium, rosuvastatin calcium, bezafibrate, atorvastatin calcium, fluvastatin sodiu, gemfibrazil, lovastatin and simvastatin) illegally added in health food products for regulating blood lipid by high performance liquid chromatography (HPLC). Methods The sample was extracted by methanol. A Inertsil ODS-3 chromatographic column was used with the mobile phase of 0.02 mol/l ammonium acetate-methanol for gradient elution, the flow rate was 1.0 ml/min, the detection wavelength was 230 nm, and the column temperature was 30. The 9 kinds of drugs were determined by the retention time, and quantified by the external standard method. Results The 9 kinds of drugs (nicotinic acid, pravastatin sodium, rosuvastatin calcium, bezafibrate, atorvastatin calcium, fluvastatin sodiu, gemfibrazil, lovastatin and simvastatin) could be analyzed qualitatively and quantitatively within 20 min. The drugs were detected at the range between 1.0 μg/ml and 100 μg/ml with good linearities, and the correlation coefficients were all greater than 0.999. Except for atorvastatin calcium in the teabag, the average recoveries were between 77.9% * 通 讯 作 者 : 珺,, E-mail: felicity81612@163.com *Corresponding author: ZHANG Hua-Jun, Competent Pharmacist, Beijing Institute for Drug Control, Beijing 102206, China. E-mail: felicity81612@163.com
8, : 9 3099 and 102.9%, and the relative standard deviations (RSDs) were 0.42%~5.8%. Conclusion The method has good linear relationships, wide linear ranges, high accuracy and precision, and can be used for the simultaneous determination of above 9 kinds of drugs. KEY WORDS: high performance liquid chromatography; health food products for regulating blood lipid; illegally added drugs 1 引 言 [1], [2],, [3],,,, [4-8] 2010 3 [9], 3 (high performance liquid chromatography, HPLC) [9] - (liquid chromatography tandem mass spectrometry, LC-MS/MS) [9-14], 3 9 2 材 料 与 方 法 2.1 仪 器 与 试 剂 1100 ( G1311A G1322A G1315B G1329A G1316A, ); SB-25-12D ( ) ( 100434-201102, 99.9%) ( 101028-201202, 97.6%) ( 100732-200501, 100%) ( 100590-201303, 95.3%) ( 100800-201302, 95.5%) ( 100284-200602, 99.5%) ( 100600-201003, 99.4%) ( 100601-201003, 99.0%) ; ( 8-XJZ-27-1, 98.0%) Toronto Research Chemical Inc (, ); ( L290P82, 98%, ); ( ) 2.2 实 验 方 法 2.2.1 色 谱 条 件 : Inertsil ODS-3(250 mm 4.6 mm, 5 μm); : 0.02 mol/l (A)- (B); : 0~5 min, 60%A~20%A; 5~10 min, 20%A; 10~20 min, 20%A~10%A; 20~21 min, 10%A~60%A; 21~30min, 60%A; : 1.0 ml/min; : 230 nm; : 30 ; : 10 μl 2.2.2 标 准 溶 液 的 制 备 10.0 mg, 10 ml, 1.0 mg/ml, ( 1.0 5.0 10.0 20.0 50.0 100.0 μg/ml) 2.2.3 供 试 品 溶 液 的 制 备 1, 50 ml, 40 ml, 15 min,,,, 0.45 μm, 3 结 果 与 分 析 3.1 分 析 条 件 的 优 化, - 0.1% - 0.02 mol/l ( 0.1% )- ; 0.02 mol/l -,,, 200~400 nm,, 9,, 230 nm
3100 7,, 230 nm,, 3.2 系 统 适 用 性 实 验, 1 3.07 8.66 9.05 9.66 11.47 13.97 15.42 17.43 18.58 min, 1.5, 5000, 3.3 线 性 关 系 定 量 限 与 检 出 限, 2.2.1,, (signal-noise ratio, S/N) 10 (limit of quantification, LOQ), S/N=3 (limit of detection, LOD), 9 1.0~100 μg/ml, 0.999, 9 1 1 Fig. 1 High performance liquid chromatogram of mixed reference 1. (nicotinic acid), 2. (pravastatin sodium), 3. (rosuvastatin calcium), 4. (bezafibrate), 5. (atorvastatin calcium), 6. (fluvastatin sodiu), 7. (gemfibrazil), 8. (lovastatin),9. (simvastatin) Table 1 表 1 9 种 化 学 药 物 的 回 归 方 程 相 关 系 数 线 性 范 围 定 量 限 及 检 出 限 Regression equations, correlation coefficients, linear ranges, LOQs and LODs of the 9 kinds of drugs (r 2 ) (μg/ml) LOQ(μg/mL) LOD(μg/mL) (nicotinic acid) Y=12.445X+3.4857 0.9998 1.0~100 0.45 0.130 (pravastatin sodium) Y=25.2243X+6.2387 0.9998 1.0~100 0.22 0.065 (rosuvastatin calcium) Y=18.179X+3.6867 0.9999 1.0~100 0.30 0.090 (bezafibrate) Y=38.645X+34.971 0.9998 1.0~100 0.15 0.046 (atorvastatin calcium) Y=21.897X-3.3620 0.9999 1.0~100 0.50 0.150 (fluvastatin sodiu) Y=27.834X-7.1343 0.9999 1.0~100 0.51 0.150 (gemfibrazil) Y=14.509X+21.132 0.9996 1.0~100 0.56 0.170 (lovastatin) Y=30.878X+0.093442 0.9999 1.0~100 0.30 0.089 (simvastatin) Y=46.068X+7.9714 0.9998 1.0~100 0.21 0.062
8, : 9 3101 3.4 准 确 度 和 精 密 度 实 验 9 : A ( : ABJ501085A) B ( : Z3A006) 3 ( 1.6 0.4 0.08 mg/g),, 6, 2,, 77.9%~102.9%, (relative standard deviation, RSD) 0.42%~5.8%,, [15],,,,, Table 2 表 2 空 白 样 品 中 9 种 化 学 药 物 的 添 加 回 收 率 及 精 密 度 (n=6) Recoveries and precisions of 9 kinds of drugs spiked in a blank matrix (n=6) (mg/g) % RSD% % RSD% (nicotinic acid) 1.60 98.4 1.20 101.5 1.70 0.40 100.9 0.93 93.0 3.10 0.08 84.6 3.20 94.8 3.50 1.60 98.4 1.80 96.8 0.67 (pravastatin sodium) 0.40 98.6 0.79 95.8 1.10 0.08 89.8 0.42 87.7 1.20 1.60 101.7 1.80 99.9 0.72 (rosuvastatin calcium) 0.40 102.2 0.77 99.9 0.93 0.08 96.8 0.96 92.9 1.30 1.60 97.2 1.80 95.3 0.55 (bezafibrate) 0.40 95.9 1.10 94.3 0.49 0.08 77.9 4.20 80.9 1.90 1.60 97.9 1.80 40.6 9.30 (atorvastatin calcium) 0.40 94.4 0.77 33.7 2.60 0.08 89.5 0.97 32.1 7.70 1.60 95.6 1.90 101.6 0.97 (fluvastatin sodiu) 0.40 95.4 0.74 102.9 0.58 0.08 86.6 1.30 96.2 0.93 1.60 99.7 1.40 94.1 3.60 (gemfibrazil) 0.40 101.2 1.10 98.4 5.80 0.08 96.7 2.20 94.1 4.20 1.60 95.5 1.80 96.2 0.92 (lovastatin) 0.40 95.7 0.81 98.3 0.66 0.08 93.3 0.53 95.8 1.10 1.60 93.3 1.90 96.5 0.87 (simvastatin) 0.40 93.5 2.40 96.1 1.10 0.08 83.9 1.10 88.5 4.30
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