目的 采用高效液相色譜-四極桿飛行時間串聯(lián)質(zhì)譜技術(shù)（HPLC-Q-TOF-MS/MS）對咳喘寧顆粒化學(xué)成分進行分析，并建立 HPLC 多成分含量測定方法。方法 采用 HPLC-Q-TOF-MS/MS 技術(shù)，色譜柱 Agilent ZORBAX C₁₈（150 mm×4.6 mm，5 μm），柱溫 35 ℃，體積流量 1.0 mL·min^-1，進樣量 10 μL，流動相 A：0.1% 甲酸，流動相 B：乙腈，梯度洗脫；電噴霧離子源（ESI），正、負(fù)離子模式掃描，結(jié)合對照品和文獻數(shù)據(jù)鑒定咳喘寧顆粒中的化學(xué)成分，并建立HPLC法同時測定新綠原酸、綠原酸、3，5-O-二咖啡?；鼘幩?、異綠原酸B、4，5-O-二咖啡酰基奎寧酸、柚皮苷的含量。色譜條件：以十八烷基硅烷鍵合硅膠為填充劑（Waters AtlantisTM T3色譜柱，250 mm×4.6 mm，5 μm），以甲醇-0.1%甲酸為流動相，梯度洗脫：0～20 min，3%～15% 甲醇；20～25 min，15%～18% 甲醇；25～26 min，18%～23% 甲醇；26～45 min，23%甲醇；45～46 min，23%～38% 甲醇；46～60 min，38% 甲醇；60～61 min，38%～42% 甲醇；61～70 min，42% 甲醇 ；70～80 min，42%～90%甲醇；體積流量1.0 mL·min^-1，柱溫30 ℃；進樣量10 μL，檢測波長324、283 nm。結(jié)果 共鑒定咳喘寧顆粒中86個化合物，20個來自生麻黃、15個來自白屈菜、30個來自金銀花、22個來自枇杷葉、12個來自金沸草、14個來自化橘紅，其中有26個化合物經(jīng)與對照品比對確認(rèn)，選擇新綠原酸、綠原酸、3，5-O-二咖啡?；鼘幩?、異綠原酸B、4，5-O-二咖啡?；鼘幩岷丸制ぼ兆鳛楹繙y定的質(zhì)控指標(biāo)，6種成分在各自質(zhì)量濃度范圍內(nèi)線性關(guān)系良好（r≥0.999 9），精密度、穩(wěn)定性及重復(fù)性良好，加樣回收率為99.48%～103.39%，RSD為1.29%～1.98%。對3批咳喘寧顆粒進行多成分含量測定，方法可行。結(jié)論 在質(zhì)譜成分解析的基礎(chǔ)上，建立咳喘寧顆粒6種化學(xué)成分的含量測定方法，為咳喘寧顆粒的藥效物質(zhì)基礎(chǔ)和質(zhì)量標(biāo)準(zhǔn)研究提供參考。

Objective High performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (HPLC-QTOF-MS/MS) was used to analyze the chemical constituents of Ke Chuanning Granules, and a HPLC method for the determination of multi-components was established.Methods Using HPLC-Q-TOF-MS/MS technology, the chromatographic column was Agilent ZORBAX C₁₈(150 mm×4.6 mm, 5 μm), the column temperature was 35 ℃, the volume flow rate was 1.0 mL·min^-1, the injection volume was 10 μL, the mobile phase A was 0.1% formic acid, and the mobile phase B was acetonitrile, and gradient elution was performed. The chemical constituents in Kechuanning Granules were identified by electrospray ionization source (ESI), positive and negative ion mode scanning, and the contents of neochlorogenic acid, chlorogenic acid, 3,5-O- dicaffeoyl quinic acid, isochlorogenic acid B, 4, 5-O-dicaffeoyl quinic acid and naringin were determined simultaneously by HPLC. Chromatographic conditions: Octadecylsilane bonded silica gel as filler (Waters AtlantisTM T3 column, 250 mm×4.6 mm, 5 μm), methanol-0.1% formic acid as mobile phase, gradient elution: 0—20 min, 3%—15% methanol; 20—25 min, 15%—18% methanol; 25—26 min, 18%—23% methanol; 26—45 min, 23% methanol; 45—46 min, 23%—38% methanol; 46—60 min, 38% methanol; 60—61 min, 38%—42% methanol; 61—70 min, 42% methanol; 70—80 min, 42%—90% methanol. The volume flow rate is 1.0 mL·min^-1, and the column temperature is 30 ℃ ; The sample volume is 10 μL, and the detection wavelength is 324 and 283 nm.Results A total of 86 compounds in Kechuanning Granules were identified, including 20 compounds from Ephedrae Herba, 15 compounds from Chelidonii Herba, 30 compounds from Lonicerae Japonicae Flos, 22 compounds from Eriobotryae Folium, 12 compounds from Inulae Herba, and 14 compounds from Citri Grandis Exocarpium. Among 26 compounds, neochlorogenic acid, chlorogenic acid, 3, 5-O-dicaffeoyl quinic acid, isochlorogenic acid B, 4, 5-O-dicaffeoyl quinic acid and naringin were selected as quality control indicators for content determination after comparison with reference substances. The linear relationship of the six main components was good (r≥ 0.999 9) within the respective concentration ranges, and the precision, stability and repeatability were good. The recovery after sample addition was 99.48%—103.39%, and RSD was 1.29%—1.98%. It is feasible to determine the multicomponent content of three batches of Kechuanning Granules.Conclusion Based on the analysis of mass spectrometry components, a method for the determination of six chemical components in Kechuanning Granules was established, which provided reference for the study of pharmacodynamic substance basis and quality standard of Kechuanning Granules.

R284.1，R285.5

2021年國家中醫(yī)藥管理局岐黃學(xué)者項目