功能化磁性纳米颗粒用于生脉饮（党参方）中黄曲霉毒素B₁的含量测定

目的：将功能化磁性纳米颗粒2-氨基-5-巯基-1，3，4-噻二唑—（3-巯基丙基）三甲氧基硅烷—磁性纳米颗粒用于生脉饮（党参方）中黄曲霉毒素B₁的净化富集处理，建立基于功能化磁性纳米颗粒的磁性固相萃取-高效液相色谱-荧光法，测定生脉饮（党参方）中黄曲霉毒素B₁含量。方法：制备功能化磁性纳米颗粒，建立磁性固相萃取-高效液相色谱-荧光法对生脉饮（党参方）中黄曲霉毒素B₁的含量进行测定。萃取条件：磁性纳米颗粒用量150 mg，吸附搅拌时间10 min，洗脱剂二氯甲烷-丙酮（2：1），洗脱剂用量9 mL，解吸附搅拌时间8 min。色谱条件：采用C₁₈色谱柱（4.6 mm×250 mm，5 μm），流动相为乙腈-甲醇-0.1%甲酸水（20：20：60），等度洗脱；流速1.0 mL·min^-1；柱温25 ℃；荧光检测器检测，激发波长360 nm，发射波长440 nm。结果：获得了优化的磁性固相萃取条件，对建立起的方法进行方法学考察，结果表明黄曲霉毒素B₁浓度在0.3~10 ng·mL^-1范围内线性关系良好（r大于0.999 5）；加样回收率为93.8%~103.5%，检测限为0.07 ng·mL^-1，定量限为0.21 ng·mL^-1；所测9批样品黄曲霉毒素B₁含量为1.04~1.93 ng·mL^-1。结论：成功建立了基于功能化磁性纳米颗粒的磁性固相萃取-高效液相色谱-荧光法，并将该法用于生脉饮（党参方）中黄曲霉毒素B₁的含量测定。本研究为建立一种低成本检测中成药中黄曲霉毒素的方法提供了依据，为更好地保障中药安全使用提供了技术支持。

Objective:Functional magnetic nanoparticles,2-amino-5-mercapto-1,3,4-thiadiazole-(3-mercaptopropyl)-rimethoxysilane-Fe₃O₄ magnetic nanoparticles(AMT-TMSPT-MNPs),were used to enrich aflatoxin B₁ in Shengmai decoction(Dangshen prescription).And then,a method by using magnetic solid phase extraction(MSPE)-high performance liquid chromatography(HPLC)-fluorescence detection(FD)was established for quantitative analysis of aflatoxin B₁ in Shengmai decoction(Dangshen prescription). Methods:Functional magnetic nanoparticles were prepared,and then a MSPE-HPLC-FD method was established for quantitative analysis of aflatoxin B₁ in Shengmai decoction(Dangshen prescription).Magnetic solid phase extraction parameters were as follows:amount of AMT-TMSPT-MNPs was 150 mg,stir time for adsorption was 10 min,eluent solvent was CH₂Cl₂-Me₂CO(2:1),elution volume was 9 mL,stir time for elution was 8 min.Chromatographic separation was performed on a C₁₈ column(4.6 mm×250 mm,5 μm)and the mobile phase was acetonitrile-methanol-0.1% formic acid(20:20:60)at a flow rate of 1.0 mL·min^-1.The column temperature was 25 ℃.The excitation and emission wavelength of the fluorescence detector was 360 nm and 440 nm,respectively.Results:Optimized magnetic solid phase extraction condition was obtained.The established MSPE-HPLC-FD method was validated.The results showed that aflatoxin B₁ had a good linear relationship in the range of 0.3-10 ng·mL^-1(r≥0.999 5);the recovery rate was 93.8%-103.5%,the limit of detection(S/N=3)was estimated to be 0.07 ng·mL^-1 and the limit of quantitation(S/N=10)was estimated to be 0.21 ng·mL^-1.The content of aflatoxin B₁ in 9 batches of samples was 1.04-1.93 ng·mL^-1.Conclusion:A method using MSPE-HPLC-FD was developed successfully for determination of aflatoxin B₁ in Shengmai decoction(Dangshen prescription).This study laid a foundation for the development of a low-cost method for determination of aflatoxin in Chinese patent medicine.It also provided technical support for guarantee of the safe use of Chinese medicine.