目的 建立HPLC-一測多評（QAMS）法同時檢測乳寧顆粒中柴胡皂苷a、柴胡皂苷d、二氫丹參酮Ⅰ、隱丹參酮、丹參酮Ⅰ、丹參酮Ⅱ_A、王不留行環(huán)肽A、王不留行環(huán)肽B的含量。方法 采用Agilent HC-C₁₈（250 mm×4.6 mm，5 μm）色譜柱，柱溫30℃；乙腈-0.1%磷酸為流動相，梯度洗脫，體積流量1.0 mL/min，檢測波長分別為210 nm（檢測柴胡皂苷a和柴胡皂苷d）和280 nm（檢測二氫丹參酮Ⅰ、隱丹參酮、丹參酮Ⅰ、丹參酮Ⅱ_A、王不留行環(huán)肽A和王不留行環(huán)肽B）。以丹參酮Ⅱ_A為內(nèi)參物，建立其與柴胡皂苷a、柴胡皂苷d、二氫丹參酮Ⅰ、隱丹參酮、丹參酮Ⅰ、王不留行環(huán)肽A、王不留行環(huán)肽B的相對校正因子（RCF），計(jì)算各成分含量，同時與外標(biāo)法（ESM）實(shí)測值進(jìn)行比較，以驗(yàn)證HPLC-QAMS法的可行性。結(jié)果 柴胡皂苷a、柴胡皂苷d、二氫丹參酮Ⅰ、隱丹參酮、丹參酮Ⅰ、丹參酮Ⅱ_A、王不留行環(huán)肽A、王不留行環(huán)肽B分別在4.08～102.00、2.97～74.25、0.74～18.50、1.56～39.00、1.98～49.50、3.69～92.25、0.66～16.50、0.54～13.50 μg/mL（r≥0.999 1）線性關(guān)系良好，平均加樣回收率（RSD）分別為100.08%（0.54%）、99.68%（0.67%）、97.85%（1.64%）、98.99%（0.92%）、98.81%（1.35%）、100.06%（0.62%）、96.81%（0.73%）、97.79%（1.41%），乳寧顆粒中各成分一測多評法計(jì)算值與外標(biāo)法實(shí)測含量值無顯著性差異。結(jié)論 所建立的HPLC-QAMS法為乳寧顆粒提供了一種準(zhǔn)確可行的多指標(biāo)質(zhì)量控制模式。

Objective To establish an HPLC-QAMS method for the determination of saikosaponin a, saikosaponin d, 15, 16-dihydrotanshinone I, cryptotanshinone, tanshinone I, tanshinone II_A, segetalin A and segetalin B in Runing granule. Methods The samples were separated on an Agilent HC-C₁₈ (250 mm×4.6 mm,5 μm),gradient elution conditions were the mobile phase using acetonitrile-0.1% phosphoric acid solution at a flow rate of 1.0 mL/min.The detection wavelength were 210 nm for saikosaponin a and saikosaponin d,and 280 nm for 15,16-dihydrotanshinone I,cryptotanshinone,tanshinone I,tanshinone II_A,segetalin A and segetalin B,the column temperature was 30 ℃.Using tanshinone II_A as an internal standard,the relative correction factors of saikosaponin a, saikosaponin d, 15, 16-dihydrotanshinone I, cryptotanshinone, tanshinone I, segetalin A and segetalin B were calculated, after which the content determination was made.The method was validated by comparison of the quantitative results between external standard method (ESM) and HPLC-QAMS method. Results Saikosaponin a,saikosaponin d, 15, 16-dihydrotanshinone I, cryptotanshinone, tanshinone I, tanshinone II_A, segetalin A and segetalin B showed good linear relationships within the ranges of 4.08 — 102.00, 2.97 — 74.25, 0.74 — 18.50, 1.56 — 39.00, 1.98 — 49.50, 3.69 — 92.25, 0.66 — 16.50 and 0.54 — 13.50 μg/mL (r ≥ 0.999 1), whose average recoveries (RSD) were 100.08% (0.54%), 99.68% (0.67%), 97.85% (1.64%), 98.99% (0.92%), 98.81% (1.35%), 100.06%(0.62%),96.81% (0.73%), 97.79%(1.41%), respectively. No significant differences were found in the quantitative analysis of components by ESM and HPLC-QAMS method. Conclusions The HPLC-QAMS method can provide a accurate and feasible method with determination to establish multi-index component quality evaluation model for Runing granule.

山東省中醫(yī)藥科技發(fā)展計(jì)劃項(xiàng)目（2019-0449）