目的 旨在开展国家卫生健康委员会和国家中医药管理局推荐治疗新型冠状病毒肺炎（COVID-19）的协定通用处方清肺排毒汤的主要化学成分及小鼠组织分布研究，为其药效物质研究提供参考依据。方法 采用UHPLC-Q-Orbitrap HRMS对清肺排毒汤的主要化学成分进行分析，色谱分离采用Acquity UPLC^® BEH C₁₈色谱柱（100 mm×2.1 mm，1.7 μm），流动相为甲醇-0.1%甲酸水，梯度洗脱，体积流量0.3 mL/min。基于化合物的一级、二级质谱信息，结合对照品比对确认，实现复方主要化学成分的快速、精准识别。在此基础上，对小鼠ig清肺排毒汤后血及组织中的移行成分进行了鉴定，并建立了同步测定麻黄碱、伪麻黄碱、苦杏仁苷、野黑樱苷、甘草苷、金丝桃苷、橙皮苷、黄芩苷、次野鸢尾黄素的定量分析方法，对小鼠ig清肺排毒汤后上述9种入血成分的组织分布进行研究。结果 从清肺排毒汤中共鉴定出39种化学成分，ig给药后，在小鼠血清、心、肝、脾、肺、肾中分别鉴定出12、9、9、8、10、10种化学成分。发现9种化学成分可快速吸收并分布到多个组织，在0.5 h时血清及各组织中除黄芩苷之外的8种化学成分的含量均达到峰值，黄芩苷含量峰值出现在2 h或4 h。重点关注的肺组织中0.5 h时各成分的暴露量依次为麻黄碱（2 759.11±784.39）ng/g > 野黑樱苷（1 819.7±427.28）ng/g > 伪麻黄碱（880.60±287.97）ng/g > 苦杏仁苷（304.43±234.70）ng/g > 橙皮苷（78.33±38.38）ng/g > 次野鸢尾黄素（8.62±4.66）ng/g > 黄芩苷（8.53±1.91）ng/g > 金丝桃苷（7.72±1.63）ng/g > 甘草苷（7.68±5.19）ng/g；2 h时成分含量为麻黄碱（776.61±148.40）ng/g > 野黑樱苷（325.20±104.17）ng/g > 伪麻黄碱（212.21±44.63）ng/g > 黄芩苷（71.72±23.96）ng/g > 苦杏仁苷（46.39±36.45）ng/g > 橙皮苷（13.39±10.99）ng/g > 金丝桃苷（3.11±0.75）ng/g > 甘草苷（2.86±0.46）ng/g；4 h时各成分质量分数为麻黄碱（327.61±212.59）ng/g > 野黑樱苷（173.77±58.21）ng/g > 伪麻黄碱（84.68±59.04）ng/g > 黄芩苷（49.33±17.06）ng/g > 苦杏仁苷（1.26±0.26）ng/g。结论 研究建立的UHPLC-Q-Orbitrap HRMS方法可实现清肺排毒汤中多种化学成分的快速、准确分析，并明确了ig给药后清肺排毒汤主要成分在小鼠的组织分布情况，为后续清肺排毒汤的药效物质基础研究及进一步的临床应用提供了药动学信息和指导。

Objective Qingfei Paidu Decoction is recommended as therapies of corona virus disease 2019 (COVID-19) by National Health Commission of the People's Republic of China and National Administration of Traditional Chinese Medicine. The purpose of this study is to investigate the main chemical constituents in Qingfei Paidu Decoction and the tissues distribution of major absorpted ingredients in mice, as well as to provide some guidance for study of its pharmacodynamic substance. Methods In this study, UHPLC-Q-Orbitrap HRMS was used to analyze the main chemical constituents in Qingfei Paidu Decoction. An Acquity UPLC^® BEH C₁₈ chromatographic column (100 mm×2.1 mm, 1.7 μm) was used as solid phase, while the mobile phase was methanol and 0.1% formic acid water. The major constituents in this Chinese medicine compound were quickly and accurately identified, via comparison with the MS and MS/MS spectra of the standards. After then, the constituents absorpted into mice blood and tissues after oral administration of Qingfei Paidu Decoction were studied. Meanwhile, an LC-MS based analytical method was established for simultaneous determination of ephedrine, pseudoephedrine, amygdalin, prunasin, liquiritin, hyperoside, hesperidin, baicalin, and risflorentin in biological samples. The tissue distribution of the nine target constituents in mice after oral administration of Qingfei Paidu Decoction were then investigated. Results A total of 39 chemicals were identified from Qingfei Paidu Decoction. After oral administered Qingfei Paidu Decoction in mice, 12, 9, 9, 8, 10, and 10 constituents were identified in serum, heart, liver, spleen, lung, and kidney, respectively. It was also found that these nine constituents could be quickly absorbed into circulation system and then distributed to various tissues. Except baicalin, the exposure of other eight target constituents were peaked in serum and tissues at 0.5 h. The exposure of baicalin was peaked at 2 h or 4 h. At 0.5 h, the exposure of target components to lung tissue were ranked as follows:ephedrine (2 759.11±784.39 ng/g) > prunasin (1 819.70±427.28 ng/g) > pseudoephedrine (880.60±287.97 ng/g) > amygdalin (304.43±234.70 ng/g) > hesperidin (78.33±38.38 ng/g) > risflorentin (8.62±4.66 ng/g) > baicalin (8.53±1.91 ng/g) > hyperoside (7.72±1.63 ng/g) > liquiritin (7.68±5.19 ng/g). At 2 h, ephedrine (776.61±148.40 ng/g) > prunasin (325.20±104.17 ng/g) > pseudoephedrine (212.21±44.63 ng/g) > baicalin (71.72±23.96 ng/g) > amygdalin (46.39±36.45 ng/g) > hesperidin (13.39±10.99 ng/g) > hyperoside (3.11±0.75 ng/g) > liquiritin (2.86±0.46 ng/g). At 4 h, ephedrine (327.61±212.59 ng/g) > prunasin (173.77±58.21 ng/g) > pseudoephedrine (84.68±59.04 ng/g) > baicalin (49.33±17.06 ng/g) > amygdalin (1.26±0.26 ng/g). Conclusion A UHPLC-Q-Orbitrap HRMS based method has been established for rapid and accurate identification of the constituents in Qingfei Paidu Decoction, while tissue distribution of the major absorpted constituents were investigated in mice following oral administration of Qingfei Paidu Decoction. These findings provided key information and guidance for further studies on pharmacodynamic substances and clinical applications of Qingfei Paidu Decoction.

R284.1

国家重点研发计划资助项目（2020YFC0845400）；国家重点研发计划资助项目（2017YFC1700200）；国家自然科学基金资助项目（81530101）；国家自然科学基金资助项目（81703681）