保健食品中添加小檗碱的检测方法研究

Detection of berberine illegally adulterated in dietary supplement

目的:建立降糖类保健食品中非法添加小檗碱的检测及含量测定方法。方法:采用薄层色谱法,以乙酸乙酯-丁酮-甲酸-水(15∶7∶1∶1)为展开剂,置紫外光灯(365 nm)下检视;采用高效液相色谱法,使用Kromasil C18色谱柱(4.6 mm×250 mm,5μm),以甲醇-离子对溶液(庚烷磺酸钠0.5056 g,加三乙胺1.4 mL,加水至1000 mL,用磷酸调pH至3.5±0.05)(60∶40)为流动相,流速0.8 mL·min-1,检测波长为348 nm;液相色谱-质谱联用法采用电喷雾电离,正离子检测模式进行检测,一级全扫描及二级选择离子扫描。结果:薄层色谱鉴别,青藤碱在365 nm下斑点清晰,Rf值适中;液相色谱法测定,小檗碱浓度在3.435～69.70μg·mL-1范围内线性关系良好(r=0.9998),平均加样回收率在101.2%,能够满足日常检验筛查的要求。结论:本研究建立的方法选择性强,灵敏度高,可用于检测保健食品中非法添加的小檗碱。

Objective： To establish TLC, HPLC and LC - MS/MS methods for the detection of the illegal berberine adulteration in dietary supplement. Methods： Acetic ether - 2 - butanone - formic acid - water （ 15 ： 7 ： 1 ： 1 ） was used as the developing solvent in the TLC ,and examined under ultraviolet light（365 nm）. HPLC method was car- ried out using a Kromasil C18 column（4. 6 mm ×250 mm, 5 μm） and detected at 348 nm; The mobile phase was a mixture of acetonitrile - ion pair reagent （ sodium 1 - heptanesulfonate 0. 5056 g, containing 1.4 mL triethyl- amine, pH was adjusted to 3.5 ±0. 05 using phosphoric acid）, and the flow rate was 0. 8 mL .min-1 LC - MS method with ESI interface was employed and operated in positive ion mode in primary full and secondary selected ion scans. Results： In the TLC, the spot of berberine at 365 nm was clear, and the Rf value was appropriate. The linear range was 3.435 -69. 70 μg . mL-l with a good correlation（r = 0. 9998）, and the average recovery was 101.2%. Conclusion ：The method is selective and sensitive, which can be used for the detection of illegal berber- ine adulteration in dietary supplement.