目的 建立芩百清肺浓缩丸（Qinbai Qingfei Concentrated Pills，QQCP）的质量控制方法。方法 正交试验优化QQCP中总生物碱、总黄酮、总皂苷的提取工艺；采用超高效液相色谱/四级杆-静电场轨道阱质谱（UHPLC/Q-Exactive Orbitrap MS）检测，分别对液相色谱流动相、柱温、梯度洗脱条件，质谱分辨率、喷雾电压等条件进行优化，确定3类成分的检测条件，依据色谱保留时间、精确质荷比、串联质谱数据，实现化学成分定性检测；对13个经对照品验证结构的化学成分进行定量方法学考察，并测定其在3批次QQCP中的含量。结果 总生物碱的最优提取工艺为料液比1∶10、40 ℃、60%乙醇、超声提取4次、每次40 min，总黄酮的最优提取工艺为料液比1∶400、70 ℃、50%乙醇、超声提取1次、50 min，总皂苷的最优提取工艺为料液比1∶30、60 ℃、40%甲醇、超声提取1次、50 min；按最优工艺获得的QQCP提取液中，共定性检测到72个化学成分，其中对叶百部碱、芥酸酰胺、硬脂酰胺、白杨素、槲皮素、汉黄芩素、千层纸素A、黄芩苷、野黄芩苷、葛根素、去芹糖桔梗皂苷D、桔梗皂苷D、去芹糖桔梗皂苷D3实现了定量检测，线性关系曲线的R²均大于0.995；以上成分在3批次QQCP中的质量分数分别为10.543～36.891、78.764～124.300、0.110～0.364、3.353～8.052、0.099～0.152、2.062～3.201、0.821～1.981、6.323～12.421、0.832～1.291、3.210～5.461、0.120～0.189、7.551～9.443、0.872～1.423 μg/g。结论 所建立的UHPLC/Q-Exactive Orbitrap MS检测方法可为QQCP的质量控制及相关化合物的定量分析提供方法学依据。

Objective To establish a quality control method for Qinbai Qingfei Concentrated Pills (QQCP). Methods Orthogonal test was conducted to optimize the extraction technology of total alkaloids, total flavonoids and total saponins in QQCP. Ultra-high performance liquid chromatography/quadrupole-exactive orbitrap mass spectrometry (UHPLC/Q-Exactive Orbitrap MS) was used to detect the above three types of components after optimizing the detection conditions including liquid chromatography mobile phase, column temperature, gradient elution conditions, mass spectrometry resolution, spray voltage, etc. Components were qualified according to the retention time, accurate m/z and MS² spectrum. Quantitative methodology was investigated based on 13 components which was confirmed by standards, and applied to quantify the targeted components in three batches of QQCP. Results The optimal extraction process for total alkaloids is liquid-to-solid ratio of 1:10, 40 ℃, 60% ethanol, ultrasonic extraction for four times, 40 min/time; that for total flavonoids is liquid-to-solid ratio of 1:400, 70 ℃, 50% ethanol, ultrasonic extraction for 1 time, 50 min; that for total saponins is liquid-to-solid ratio of 1:30, 60 ℃, 40% methanol, ultrasonic extraction for 1 time, 50 min. There were total of 72 components which were qualified. A quantitative method, with satisfactory linearity and R² > 0.995, for 13 components which contained tuberostemonine, erucamide, stearamide, chrysin, quercetin, wogonin, oroxylin A, baicalin, scutellarin, puerarin, deapioplatycodin D, platycodin D and deapioplatycodin D₃, were established. Their content in three batches of QQCP were 10.543—36.891, 78.764—124.300, 0.110—0.364, 3.353—8.052, 0.099—0.152, 2.062—3.201, 0.821—1.981, 6.323—12.421, 0.832—1.291, 3.210—5.461, 0.120—0.189, 7.551—9.443, 0.872—1.423 μg/g, respectively. Conclusion The established UHPLC/ Q-Exactive Orbitrap MS method can provide methodological basis for the quality control of QQCP as well as the quantitative analysis of related compounds.

R283.6

国家自然科学基金资助项目(82174060);黑龙江省省属高等学校基本科研业务费科研项目(145209505);黑龙江省博士后资助项目(LBH-Z19099)