HPLC-柱后光化学衍生法测定清火片(胶囊)中黄曲霉毒素G_2、G_1、B_2、B_1的含量及其安全性评价

目的:建立测定清火片(胶囊)中黄曲霉毒素(AF)G_2、G_1、B_2、B_1的方法,并评价该制剂的安全性。方法:采用高效液相色谱(HPLC)-柱后光化学衍生法,并以全国37个厂家生产的266批清火片(胶囊)为样品。色谱柱为Agilent C₁₈;流动相为水-乙腈-甲醇(V/V/V),梯度洗脱;流速为1.0 mL/min;柱温为40℃;进样量为10μL;荧光检测器激发波长为360 nm、发射波长为450 nm。结果:AF G_2、AF G_1、AF B_2、AF B_1分别在进样量为10.197～101.97(r=0.999 7)、10.197～101.97(r=0.999 6)、9.958 6～99.586(r=0.999 1)、9.999 0～99.990(r=0.998 3)pg范围内线性关系良好;精密度(n=6)、重复性(n=6)、稳定性(12 h,n=5)试验的RSD均小于3.0%;检测限分别为0.80、4.00、0.80、4.00 pg;定量限分别为1.60、8.00、1.60、8.00 pg;加样回收率分别为85%～90%、85%～90%、55%～65%、65%～75%(RSD为1.8%～4.7%,n=6)。在266批样品中均未检测出AF G_2、AF G_1、AF B_2、AF B_1。结论:该方法可用于清火片(胶囊)中AF的检测;虽在抽检样品中未检测出AF,但建议增订AF检查项,以提高其安全性。

Content Determination of Aflatoxin G_2,G_1,B_2 and B_1 in Qinghuo Tablets(Capsules) by HPLC-post-column Photochemical Derivatization and Its Safety Evaluation

OBJECTIVE:To establish a method for content determination of aflatoxin(AF)G2,G1,B2 and B1 in Qinghuo tablets(capsules),and to evaluate the safety of the preparation.METHODS:HPLC-post-column photochemical derivatization was adopted,and 266 batches of Qinghuo tablets(capsules)from 37 manufacturers as sample.The determination was performed on Agilent C18 column with mobile phase consisted of water-acetonitrile-methanol(V/V/V,gradient elution)at the flow rate of 1.0 mL/min.The column temperature was set at 40 ℃.Excitation wavelength and emission wavelength of fluorescence detector were 360 and 450 nm.RESULTS:The linear ranges of AF G2,AF G1,AF B2 and AF B1 were 10.197-101.97(r=0.999 7),10.197-101.97(r=0.999 6),9.958 6-99.586(r=0.999 1),9.999 0-99.990(r=0.998 3)pg,respectively.RSDs of precision(n=6),reproducibility(n=6)and stability tests(12 h,n=5)were all lower than 3.0%.The detection limits were 0.80,4.00,0.80 and 4.00 pg,respectively.The quantitation limits were 1.60,8.00,1.60 and 8.00 pg,respectively.The recoveries were 85%-90%,85%-90%,55%-65%,65%-75%(RSD=1.8%-4.7%,n=6).AF G2,AF G1,AF B2 and AF B1 were not detected in 266 batches of samples.CONCLUSIONS:This method is suitable for the determination of AF in Qinghuo tablets(capsules).Although AF was not detected in the sample,it is advisable to add the determination of AF so as to improve its safety.