双冬胶囊HPLC指纹图谱建立及其抗炎作用的谱效关系研究

Establishment of HPLC fingerprint of Shuangdong capsules and the spectral effect relationship of its antiinflammatory effect

目的建立双冬胶囊高效液相色谱(HPLC)指纹图谱,并研究其抗炎作用的谱效关系。方法采用HPLC法建立15批双冬胶囊的指纹图谱,同时进行相似度评价。建立大鼠足肿胀模型考察双冬胶囊的抗炎活性,以大鼠足肿胀率及右后足组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、前列腺素E2(PGE2)、白细胞介素6(IL-6)、IL-8、IL-1β、肿瘤坏死因子α(TNF-α)水平为抗炎药效指标,采用灰色关联度分析法构建双冬胶囊抗炎作用的谱效关系。结果15批双冬胶囊确定了20个共有峰,相似度均大于0.97,指认了8个共有峰。经灰色关联度分析可知,15批双冬胶囊各共有峰峰面积与足肿胀率及MDA、SOD、PGE2、IL-6、IL-8、IL-1β、TNF-α水平的关联度分别为0.6211~0.7835、0.5643~0.8279、0.5810~0.8453、0.5649~0.8550、0.5831~0.8564、0.5765~0.8635、0.5641~0.8380、0.5725~0.8513。其中,与抗炎药效指标关联性较强的有峰4(京尼平苷酸)、峰10(绿原酸)和峰2所代表的化学成分。结论本研究成功建立15批双冬胶囊的HPLC指纹图谱,其中京尼平苷酸、绿原酸等可能是其抗炎药效成分。

OBJECTIVE To establish HPLC fingerprint of Shuangdong capsules,and to study the spectral effect relationship of its anti-inflammatory effect.METHODS The fingerprints of 15 batches of Shuangdong capsules were established by HPLC,and the similarity evaluation was carried out;the foot swelling model was established to investigate the anti-inflammatory activity of Shuangdong capsules.The gray correlation analysis method was used to construct the spectral effect relationship for the antiinflammatory effect of Shuangdong capsules using the swelling rate of rat foot and the levels of malondialdehyde(MDA),superoxide dismutase(SOD),prostaglandin E2(PGE2),interleukin-6(IL-6),IL-8,IL-1βand tumor necrosis factor-α(TNF-α)in right hindfoot tissues as the pharmacodynamic indexes of anti-inflammatory effects.RESULTS Overall 15 batches of Shuangdong capsules identified 20 common peaks,the similarities were all greater than 0.97,and a total of 8 chromatographic peaks were identified.According to the gray correlation analysis,the correlation degrees between the peak area and the foot swelling rate and the levels of MDA,SOD,PGE2,IL-6,IL-8,IL-1βand TNF-αin 15 batches of Shuangdong capsules were 0.6211-0.7835,0.5643-0.8279,0.5810-0.8453,0.5649-0.8550,0.5831-0.8564,0.5765-0.8635,0.5641-0.8380 and 0.5725-0.8513,respectively.Among them,the chemical components represented by peak 4(geniposidic acid),peak 10(chlorogenic acid)and the chemical composition represented by peak 2 were strongly correlated with anti-inflammatory efficacy indicators.CONCLUSIONS In this study,HPLC fingerprints of 15 batches of Shuangdong capsules were successfully established.Among them,geniposidic acid,chlorogenic acid may be its anti-inflammatory ingredients.