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of 10 batches of L. japonica from different production areas was established according to TCM Chromatographic Fingerprint
Similarity Evaluation System (2012 edition). By comparing with reference substance,chemical constituents corresponding to
common peaks were identified,and the similarity analysis was conducted. Acute and chronic inflammatory models of mice induced
by xylene,carrageenan and cotton ball were used to evaluate inhibition rate of 10 batches of L. japonica to ear,foot and granuloma
swelling; the average value was calculated as the comprehensive pharmacodynamic index. The spectrum-effect relationship with
HPLC fingerprint of L. japonica and anti-inflammatory effect was analyzed by grey relational analysis(GRA)and partial least
squares regressiosn(PLSR)based on common peak area and comprehensive pharmacodynamic index. Chromatographic peaks with
correlation>0.7 and regression coefficient of PLSR model>0 were characteristic peaks. The percentage of peak areas of
characteristic peaks to peak areas of common peak was calculated in 10 batches of L. japonica(e.g.“peak ratio”). RESULTS:
There were 25 common peaks in HPLC fingerprints of 10 batches of L. japonica,with similarity of 0.775-0.994. Totally 9 peaks
were confirmed,i.e. rutin(peak 18),hyperoside(peak 20),isochlorogenc acid B(peak 22),galuteolin(peak 21),chlorogenc
acid(peak 9),loganin(peak 10),neochlorogenic acid(peak 2),isochlorogenic acid C(peak 25),isochlorogenic acid A(peak
23). All 10 batches of L. japonica had inhibitory effects on ear swelling,foot swelling and granuloma,with average inhibitory rate
of 47.95%-56.52%. The correlation by GRA was peak 8>12>18>16>3>11>20>22>19>21>1>9>10>13>24>14>2>
17>25>23>5>4>15,and all of correlations were greater than 0.7. The regression coefficient of PLSR for peaks 2,4,5,7,8,
10,12,13,14,15,16,17,18,20,21,22,24 were all greater than 0;those peaks were positively correlated with
anti-inflammatory effect and were characteristic peaks except for peak 7; among them,VIP values of peaks 5,8,10,16,18,
20,24 were greater than 1. The peak ratio of 10 batches of L. japonica was 58.61%-71.19%. CONCLUSIONS:HPLC fingerprint
of 10 batches of L. Japonica is successfully established. 10 batches of samples have similar components,and the content of
anti-inflammatory components is relatively high. The proportion of characteristic peaks to common peaks should not be less than
51.8%.
KEYWORDS Lonicera japonica;HPLC;Fingerprint;Anti-inflammatory effect;Spectrum-effect relationship;Grey relational
analysis;Partial least squares
金银花 Lonicerae Japonicae Flos 为忍冬科忍冬属植 1.2 药品与试剂
物忍冬Lonicera japonica Thunb.的干燥花蕾或待初开的 10 批金银花药材(编号:S1~S10)经贵州中医药大
花,主要分布于我国贵州、河南、山东和湖北等地,其经 学生药学教研室孙庆文教授鉴定为忍冬科植物忍冬 L.
济、食用及药用价值较高,属药食同源植物 。金银花作 japonica Thunb.的干燥花蕾或待初开的花,其来源信息
[1]
为药用历史悠久的中草药,具有抗病毒、抗肿瘤、提高免 详见表1。
疫和抗氧化等诸多功效 [1-2] 。目前,已有关于金银花指纹 表1 金银花药材样品来源信息
[3]
图谱及其抗炎药效的研究,如刘惠等 建立了不同批次 Tab 1 Information of L. japonica sources
金银花的高效液相色谱(HPLC)指纹图谱,结果表明,10 编号 产地 编号 产地
S1 山东临沂 S6 河南封丘
[4]
批金银花指纹图谱的相似度均在 0.92 以上;胡璇等 对 S2 山东临沂 S7 河南密县
四倍体金银花水提物的抗炎作用进行了研究,结果表 S3 湖北罗田 S8 安徽亳州
明,该水提物具有明显的抗炎作用。但是,目前尚无研 S4 河北巨鹿 S9 河北获鹿
S5 河南新乡 S10 山东枣庄
究探讨指纹图谱与抗炎药效的直接内在联系 。基于
[5]
马 钱 苷 对 照 品(批 号 :111640-201808,纯 度 :
此,本研究采用灰色关联度分析法(GRA)与偏最小二乘
99.0%)、木犀草苷对照品(批号:111720-201609,纯度:
回归分析法(PLSR)联合建立金银花指纹图谱与抗炎药
94.9%)、绿原酸对照品(批号:110753-201817,纯度:
效的谱-效关系,以期从该药中筛选出与抗炎药效密切
96.8%)、金丝桃苷对照品(批号:111521-201708,纯度:
相关的化学成分,旨在为金银花的抗炎物质基础研究和
95.1%)、芦 丁 对 照 品(批 号 :100080-201610,纯 度 :
临床应用提供科学依据。
92.6%)均购自中国食品药品检定研究院;异绿原酸 A、
1 材料 异绿原酸 B、异绿原酸 C、新绿原酸对照品(成都植标化
1.1 仪器 纯生物技术有限公司,批号:181214、181209、190102、
1260 型 HPLC 仪(美国 Agilent 公司);AE240 型电 181209,纯度均大于 98%);蒲地蓝消炎片(安徽济人药
子分析天平[梅特勒-托利多仪器(上海)有限公司]; 业有限公司,批号:20180514,规格:每片重 0.3 g);阿司
SK8210LHC 型超声波清洗仪(上海商信化玻仪器有限 匹林肠溶片(沈阳奥吉娜药业有限公司,批号:180914,
公司);HHW21.600AⅡ型恒温水浴锅(天津市泰斯特仪 规格:100 mg);角叉菜胶(上海伊卡生物技术有限公司,
器有限公司)。 批号:EK170811,纯度:99%);生理盐水(安徽丰原药业
·2498 · China Pharmacy 2020 Vol. 31 No. 20 中国药房 2020年第31卷第20期