斑蝥HPLC数字化指纹图谱研究

Study on the digitized HPLC fingerprints of Mylabris

目的:建立斑蝥 HPLC 数字化指纹图谱控制方法。方法:采用 Centurysil C_(18)BDS 色谱柱(200 mm×4.6 mm,5μm),以1%醋酸水溶液(A)-1%醋酸乙腈溶液(B)为流动相进行梯度洗脱(0～5 min,0% B;5～11 min,0%→3%B;11～22 min,3%→6% B;22～30 min,6%→12% B;30～45 min,12%→25% B;45～60 min,25%→40% B),流速1.0 mL·min^(-1),检测波长265 nm,柱温为(30.0±0.15)℃,进样量10μL。用"中药色谱指纹图谱超信息特征数字化评价系统3.0"软件计算10批斑蝥 HPLC 指纹图谱的色谱指纹图谱指数 F 和双定性双定量相似度等46个参数进行数字化评价。以双参照物体系标定指纹峰的表观相对分子质量、峰位、洗脱动量数(δ)、折合相对积分(φ)。结果:以脲苷峰为参照物峰,确定了15个共有指纹峰,建立了斑螫及其不同部位的 HPLC 数字化指纹图谱。获得了判别斑蝥质量的重要数字化判据。以双定性双定量相似度法评价斑蝥批间质量的稳定性。结论:所建立的 HPLC 数字化指纹图谱方法具有较好的稳定性和重现性,适用于斑蝥质量控制。数字化指纹图谱是中药信息质量控制的核心技术,双定性双定量相似度法是宏观定性定量评价中药质量的理想技术。

Objective：To establish a digitized HPLC fingerprint method of Mylabris. Method：The chromatographic fingerprints were obtained by injecting 10 μL of the sample solution each time on a Century SIL C18 BDS column（200 mm × 4.6 mm,5 μm） with the gradient elution solvent system composed of 1% acetate acid solution （A）and 1% acetate acid acetonitrile（B） （0 -5 min,0% B;5-11 min,0%→3% B;11 -22 min,3%→6% B;22-30 min,6%→12% B;30-45 min,12%→25% B;45-60 min,25%→40% B）.The flow rate was 1.0 mL · min ^-1, the column temperature was maintained at（30 ± 0. 15） ℃ and the detection wavelength was set at 265 nm. The chromatographic fingerprints were characteristically digitized and evaluated by the software of the digitized evaluation system for the super information characteristics of Traditional Chinese Medicine（TCM） fingerprint to give 46 parameters such as chromatographic fingerprint index, the dual qualitative and dual quantitative similarities, which can thoroughly disclose the hidden information in the fingerprints. Meanwhile, the apparent relative molecular mass,peak position, elution force index and pseudo peak area were marked in terms of the two peaks selected as the referential peaks. Results： 15 co - possessing peaks were selected as the fingerprint peaks of Mylabris by taking uridnine peak as the referential peak to establish the digitized fingerprint of Mylabris and its different body part and obtain the important digitized information for its quality control. The quality stabilities among different lots of samples were evaluated by the dual qualitative and dual quantitative similarity method. Conclusions：This new digitized fingerprint method with good precision and reproducibility can be perfectly applied in the quality control of Mylabris. Digitized fingerprint is certainly the core techniques of digital TCM quality control, and the dual qualitative and dual quantitative similarity method is fittingly to assess and control the quality of mod-ern TCM.