干馏制鲜竹沥饮片差异成分分析及其关联的安全性评价

Analysis of Different Components and Their Associated Safety Evaluation of Succus Bambusae Prepared by Dry Distillation Process

目的通过比较干馏制鲜竹沥与传统工艺鲜竹沥饮片的物质成分差异,进一步从急性毒性和亚急性毒性角度分析干馏制鲜竹沥饮片的安全性,并将成分差异和安全性进行关联分析。为干馏制鲜竹沥大品种的工艺规范化和质量标准的进一步提升提供数据支撑。方法采用气相色谱质谱联用(GC-MS)方法、结合主成分分析(PCA)和正交偏最小乘法判别分析(OPLS-DA)数学算法,分析得到三种工艺的差异性成分以及独有成分,进一步采用HPLC方法对安全性相关指标进行定量分析。采用急性毒性试验和亚急性毒性试验相结合的方案对安全性进行评估。结果分析表明,从不同工艺鲜竹沥中共归属了47种成分,包括酚类16种、酮类11种、酯类9种、醛类7种、羧酸类3种、醇类1种,相对含量和物质种类具有明显改变。对安全性相关成分苯酚和愈创木酚定量分析结果显示,干馏与火烤工艺中均检出苯酚和愈创木酚,含量分别为(0.097±0.004)和(0.150±0.017)mg·mL^(-1)。急性毒性试验结果表明,干馏制鲜竹沥饮片对小鼠体重和胃肠道存在不良影响。亚急性毒性试验结果与急性毒性试验结果具有一致性。干馏制鲜竹沥高剂量组对胃肠道组织存在不良影响,心、肝、肺和肾脏器指数也显著升高,但低、中剂量组未产生不良效应。结论干馏制鲜竹沥饮片物质基础的不同确实存在安全性风险,且关联分析表明该风险为苯酚及愈创木酚等具有刺激性的酚类成分所致。但综合来说,安全性评价提示干馏制鲜竹沥饮片总体安全性较好。这为鲜竹沥饮片的临床应用提供了毒理学数据支撑。

OBJECTIVE To compare the differences in the chemical compositionof Succus Bambusae with different processes,analyze the safety of Succus Bambusae produced by dry distillation from theperspectives of acuteand subacute toxiciy and provide data support for the further improvement ofthe process standardization andquality standards ofthe large variety offresh bamboo distillate.METHODS Using GC-MS method,combined with PCA and OPLS-DA mathematical algorithms,the differential components and unique components among the three processes were analyzed,and the HPLC method was used to quantitatively analyze the indicators related to toxicity potential.Toxic potential was assessed using a combined acute and subacute toxicity protocol.RESULTS Relative content and substance types have changed significantly,total of 47 components were identified from different processes of Succus Bambusae,including 16 phenols,11 ketones,9 esters,7 aldehydes,3 carboxylic acids,and 1 alcohol.The results of quantitative analysis of phenol and guaiacol showed that only phenol and guaiacol were detected in the dry distillation process,and the contents were(0.097±0.004)and(0.150±0.017)mg·mL^(-1) respectively.The results of acute toxicity test showed that dry distillation of Succus Bambusae decoction had adverse effects on the body weight and gastrointestinal tract of mice.The subacute toxicity test results are consistent with the acute toxicity test results.The high-dose group of dry-distilled Succus Bambusae had adverse effects on the gastrointestinal tract tissue,and the organ indexes of heart,liver,lung and kidney were also significantly increased,but no adverse effects appeared in the low-dose and middle-dose groups.CONCLUSION The huge difference in the material basis of dry distillation Succus Bambusae does have safety risks,and the correlation analysis shows that the risk factors are irritating phenolic components such as phenol and guaiacol.However,the safety evaluation indicates that dry distillation of Succus Bambusae is generally safer.This paper could provide toxicological data support for the clinical application of Succus Bambusae and its preparations.