目的 建立連翹Forsythiae Fructus的UPLC指紋圖譜及多成分含量測(cè)定方法，并結(jié)合化學(xué)計(jì)量學(xué)方法尋找不同產(chǎn)地連翹藥材質(zhì)量差異成分。方法 采用菲羅門Titank C₁₈色譜柱（150 mm×2.1 mm，1.8 μm），以體積分?jǐn)?shù)乙腈（A）-0.2%甲酸水溶液（B）為流動(dòng)相，梯度洗脫（0～5 min，5%～12% A；5～8 min，12%～20% A；8～15 min，20% A；15～18 min，20%～29% A；18～20 min，29%～48% A；20～25 min，48%～70% A；25～28 min，70%～85% A；28～30 min，85%～5% A），體積流量為0.3 mL/min，柱溫為30 ℃，進(jìn)樣量為0.4 μL，采用定時(shí)波長(zhǎng)檢測(cè)（0～13.2 min，265 nm；13.2～20 min，260 nm；20～21.5 min，270 nm；21.5～22 min，275 nm；22～30 min，280 nm）。采用中藥指紋圖譜相似度評(píng)價(jià)軟件進(jìn)行相似度評(píng)價(jià)，結(jié)合化學(xué)計(jì)量學(xué)分析，同時(shí)測(cè)定連翹酯苷A、蘆丁、連翹苷、松脂醇、連翹脂素的含量。結(jié)果 建立了連翹藥材的指紋圖譜，以6號(hào)峰為參照峰，共標(biāo)記了11個(gè)共有峰，用對(duì)照品比對(duì)法指認(rèn)出5個(gè)色譜峰，分別為峰6（連翹酯苷A）、峰7（蘆?。?、峰9（連翹苷）、峰10（松脂醇）、峰11（連翹脂素）。24批連翹藥材樣品指紋圖譜相似度均在0.99以上。聚類分析將24批連翹樣品分為3類，不同產(chǎn)地來源各自聚為一類。主成分分析表明，不同產(chǎn)地來源的連翹樣品間存在差異。連翹酯苷A、蘆丁、連翹苷、松脂醇、連翹脂素的質(zhì)量分?jǐn)?shù)分別為59.286 5～114.849 2、2.653 6～4.055 6、7.133 3～11.911 1、0.300 7～1.614 6、0.089 3～0.838 2 μg/g，經(jīng)方法學(xué)考察，各成分呈現(xiàn)良好的線性關(guān)系，平均加樣回收率在104.30%～111.48%，RSD在1.86%～2.96%。通過OPLS-DA項(xiàng)下的變量權(quán)重值（variable importance projection，VIP）分析篩選出連翹酯苷A等3個(gè)成分可作為不同產(chǎn)地連翹的差異標(biāo)志物。結(jié)論 建立的UPLC指紋圖譜和多成分含量測(cè)定方法穩(wěn)定、可靠，可為連翹藥材的質(zhì)量評(píng)價(jià)與控制提供科學(xué)依據(jù)及參考。

Objective To establish an ultra-high performance liquid chromatography (UPLC) fingerprint and multi-component content determination method for Forsythiae Fructus from different sources, and to evaluate the quality of Forsythiae Fructus from different sources in combination with chemometrics methods, so as to provide technical method reference and basic data for its quality control research. Methods Phenomenex Titank C₁₈ chromatographic column (150 mm × 2.1 mm, 1.8 μm ) was used, with acetonitrile (A) - 0.2% formic acid water (B) solution as mobile phase, gradient elution (0—5 min, 5%—12% A; 5～8 min, 12%—20% A; 8—15 min, 20% A; 15—18 min, 20%—29% A; 18—20 min, 29%—48% A; 20—25 min, 48%—70% A; 25—28 min, 70%—85% A; 28—30 min, 85%—5% A). The flow rate was 0.3 mL/min, the column temperature was 30 °C, the injection volume was 0.4 μL, and time-programmed wavelength detection wavelength was set at a fixed wavelength (0—13.2 min, 265 nm; 13.2—20 min, 260 nm; 20—21.5 min, 270 nm; 21.5—22 min, 275 nm; 22—30 min, 280 nm). The fingerprints of Forsythiae Fructus from different sources were constructed, and the contents of forsythiaside A, rutin, forsythin, pinoresinol and forsythigenin were determined by similarity analysis and chemometrics analysis. Results The fingerprint of Forsythia suspensa was established, and taking peak 6 as the reference peak, 11 common peaks were marked. A total of five chromatographic peaks were identified by reference substance comparison method, which were peak 6 (forsythiaside A), peak 7 (rutin), peak 9 (forsythin), peak 10 (pinoresinol) and peak 11 (forsythigenin). The similarity of fingerprints of 24 batches of Forsythiae Fructus samples was above 0.99. Cluster analysis divided 24 batches of Forsythiae Fructus samples into three categories, and different origins were clustered into one category. Principal component analysis showed that there were differences among Forsythiae Fructus samples from different origins. The mass fraction ranges of forsythiaside A, rutin, forsythin, pinoresinol and forsythigenin were 59.286 5—114.849 2、2.653 6—4.055 6、7.133 3—11.911 1、0.300 7—1.614 6、0.089 3—0.838 2 μg/g, respectively. The methodological investigation showed that each component showed a good linear relationship. The average recovery was between 104.30 % and 111.48 %, and the RSD was between 1.86 % and 2.96 %. Three components such as forsythiaside A were screened by VIP analysis under OPLS-DA, which could be used as differential markers of Forsythiae Fructus in different producing areas. Conclusion The UPLC fingerprint and multi-component content determination method established in this study are stable and reliable, which can provide scientific basis and reference for the quality evaluation and control of Forsythiae Fructus.

R286.2

河北省省級(jí)科技計(jì)劃項(xiàng)目資助（21372503D）