目的 建立高效液相結(jié)合一測多評（HPLC-QAMS）法同時測定金果含片中哈巴苷、哈巴俄苷、柚皮蕓香苷、橙皮苷、川陳皮素、橘紅素、麥冬甲基黃烷酮A和甲基麥冬二氫高異黃酮B的方法。方法 采用Agilent HC C18色譜柱（250 mm×4.6 mm，5 μm），流動相為乙腈-0.05%磷酸溶液，梯度洗脫；檢測波長為210 nm（哈巴苷和哈巴俄苷）和296 nm（柚皮蕓香苷、橙皮苷、川陳皮素、橘紅素、麥冬甲基黃烷酮A和甲基麥冬二氫高異黃酮B）；體積流量1.0 mL/min；進樣量10 μL；柱溫30℃。進行專屬性試驗、線性關(guān)系考察、精密度試驗、穩(wěn)定性試驗、重復(fù)性試驗、加樣回收率試驗。以橙皮苷為內(nèi)參物，建立該成分與哈巴苷、哈巴俄苷、柚皮蕓香苷、川陳皮素、橘紅素、麥冬甲基黃烷酮A和甲基麥冬二氫高異黃酮B的相對校正因子（RCF），并計算金果含片中8種成分的含量，同時與外標(biāo)法的測定結(jié)果進行比較，驗證該方法的可行性。結(jié)果 哈巴苷、哈巴俄苷、柚皮蕓香苷、橙皮苷、川陳皮素、橘紅素、麥冬甲基黃烷酮A和甲基麥冬二氫高異黃酮B檢測質(zhì)量濃度的線性范圍分別為6.54～163.50、2.27～56.75、8.16～204.00、17.95～448.75、1.88～47.00、0.54～13.50、1.48～37.00和0.76～19.00 μg/mL（r值范圍0.999 1～0.999 8）；平均加樣回收率及RSD值分別為99.85%（0.76%）、97.62%（1.07%）、100.09%（0.64%）、99.80%（0.71%）、97.48%（1.44%）、98.25%（1.60%）、96.90%（1.03%）、96.87%（1.31%），方法學(xué)驗證結(jié)果均符合要求。HPLC-QAMS法計算值與外標(biāo)法實測值無顯著性差異。結(jié)論 所建立的HPLC-QAMS法簡便、有效、準(zhǔn)確，可用于金果含片中多指標(biāo)成分含量的同時測定，為提升金果含片的質(zhì)量控制方法提供參考依據(jù)。

Objective To develop an HPLC-QAMS method for simultaneous determination of harpagide, harpagoside, narirutin, hesperidin, nobiletin, tangeretin, methylophiopogonanone A and methylophiopogonanone B in Jinguo Hanpian. Methods The determination was performed on an Agilent HC C18 column(250 mm×4.6 mm, 5 μm) with gradient elution, and the mobile phase consisted of acetonitrile-0.05% phosphoric acid at the flow rate of 1.0 mL/min. The detection wavelengths were set at 210 nm for harpagide and harpagoside, 296 nm for narirutin, hesperidin, nobiletin, tangeretin, methylophiopogonanone A and methylophiopogonanone B. The column temperature was set at 30℃, and the injection volume was 10 μL.Using hesperidin as an internal standard, the relative correction factor (RCF) of harpagide, harpagoside, narirutin, nobiletin, tangeretin, methylophiopogonanone A and methylophiopogonanone B were calculated. The RCFs value were used to calculate the contents of eight components in Jinguo Hanpian, and compared with the results of the external standard method (ESM) to verify the feasibility of the method. Results The linear range was 6.54-163.50 μg/mL for harpagide, 2.27-56.75 μg/mL for harpagoside, 8.16-204.00 μg/mL for narirutin, 17.95-448.75 μg/mL for hesperidin, 1.88-47.00 μg/mLfor nobiletin, 0.54-13.50 μg/mL for tangeretin, 1.48-37.00 μg/mL for methylophiopogonanone A, 0.76-19.00 μg/mL for methylophiopogonanone B (r=0.999 1-0.999 8). The average recovery and RSD were 99.85% (0.76%), 97.62% (1.07%), 100.09% (0.64%), 99.80% (0.71%), 97.48% (1.44%), 98.25% (1.60%), 96.90% (1.03%) and 96.87% (1.31%). Significant difference weren't found between the calculated value of HPLC-QAMS method and the measured value of external standard method. Conclusion The HPLC-QAMS is simple, effective and accurate, which can be used for simultaneous determination of multi-index component in Jinguo Hanpian, and it can provide reference for the increase of quality control method of Jinguo Hanpian.

R927.2

山東省中醫(yī)藥科技發(fā)展計劃項目（2019-0449）