一点红不同营养器官中鞣质和绿原酸含量测定(1)
摘 要:目的:建立一点红不同营养器官中鞣质和绿原酸含量测定方法。方法:以没食子酸为对照品,采用磷钼钨酸-干酪素比色法测定了鞣质含量;以绿原酸为对照品,采用高效液相色谱法测定了绿原酸含量。结果:没食子酸浓度在2～10mg•L-1范围内,与吸光度呈良好线性关系,平均加样回收率为100.32%,RSD=1.45%,一点红叶、茎、根中鞣质含量分别为0.25%、0.15%、0.22%;绿原酸含量在0.0356～0.5340μg范围内与峰面积积分值呈良好线性关系,平均加样回收率为99.37%,RSD=1.52%,一点红叶、茎、根中绿原酸含量分别为1.28%、1.14%、0.91%。结论:这两种方法灵敏,准确性强,精密度高,重复性好,适用于一点红中鞣质和绿原酸含量的测定。
关键词:一点红;鞣质;绿原酸;含量测定
中图分类号:R284.1 文献标识码:A 文章编号:1673-7717(2010)01-0125-03
Determination of Tannins and Chlorogenic Acid of Different Vegetative Organs in Emilia sonchifolia (Linn.)DC
ZHANG Yan, ZHANG Hongbin, CHEN Guangzhou,WANG Yan
(Department of Life and Science, Qiongzhou College, Wuzhishan 572200,Hainan, China)
Abstract:Objective: To establish the analytical methods for determinating the contents of tannins and chlorogenic acid of different vegetative organs in Emilia sonchifolia (Linn.)DC. Methods: The content of tannins, expressed as gallic acid equivalents, was measured by using the phospho molybdenum tungstic acid-casein colorimetry, and the content of chlorogenic acid, expressed as chlorogenic acid equivalents, was measured by HPLC. Results: The good linearity between concentrations and absorbances of gallic acid was found in the range of 2～10 mg L-1, the average recovery was 100.32% and RSD was 1.45%, the contents of tannins in leaves, stems and roots of Emilia sonchifolia (Linn.)DC were 0.25%,0.15% and 0.22% respectively; The good linearity between content and peak area integral value of chlorogenic acid was found in the range of 0.0356~0.5340μg, the average recovery was 99.37% and RSD was 1.52%, the contents of chlorogenic acid in leaves, stems and roots of Emilia sonchifolia (Linn.)DC were 1.28%,1.14% and 0.91% respectively. Conclusion: Two proposed methods are sensitive, precise, accurate and reproducible, and they are suitable for the determination of tannins and chlorogenic acid in Emilia sonchifolia (Linn.)DC.
Key words:Emilia sonchifolia (Linn.)DC; tannins; chlorogenic acid; determination
一点红[Emilia sonchifolia (Linn.)DC][1],为菊科一点红属植物,一年生或多年生草本,又名叶下红、羊蹄草、红背叶、山羊草、野芥兰、牛尾膝等,在我国南方各省均有分布。一点红全草入药[2],清热凉血,活血化瘀,利尿消肿,消炎解毒。主治上呼吸道感染、咽喉肿痛、口腔溃疡、肺炎、急性肠炎、细菌性痢疾、泌尿系统感染、睾丸炎、乳腺炎、疖肿疮疡、皮肤湿疹、跌打扭伤。它还是目前市场上畅销的妇科良药——花红冲剂的主要原料。近些年来,一点红又成为人们喜爱的一种野菜[3],食味柔滑,口感清爽。
目前对一点红的研究主要集中在部分化学成分、栽培技术等方面[4-7],对其次生代谢产物含量的研究较少。本文对一点红不同营养器官中鞣质和绿原酸含量进行了测定分析,为一点红的药用质量控制及其次生代谢产物的进一步研究和开发提供参考依据。
1 材料与试剂
1.1 材料
一点红采自海南五指山市郊(由本系林教授鉴定为一点红全草),将其洗净,分成根、茎、叶三部分,在40℃条件下烘干至恒重,粉碎,过40目筛,放在干燥器中备用。
1.2 主要仪器LC-2010c型高效液相色谱仪(LCSOLUTION工作站,日本岛津公司);BP211D电子天平(德国Sartorius公司);UV-2100双光束紫外可见分光光度计(北京瑞利分析仪器公司);CO250型超声波清洗器(上海超声仪器厂)。
1.3 试剂没食子酸对照品(中国药品生物制品检定所,批号110831-200302);绿原酸对照品(中国药品生物制品检定所,批号110753-200413);磷钼钨酸试液(按《中华人民共和国药典》2005年版一部附录ⅩⅤB配制)[8];乙腈为色谱纯;水为二次蒸馏水;其他试剂均为分析纯。
2 方法与结果
2.1 鞣质含量测定
2.1.1 对照品溶液的制备精密称取没食子酸对照品50mg,置100mL棕色量瓶中,加水溶解并稀释至刻度,精密量取5mL,置50mL棕色量瓶中,用水稀释至刻度,摇匀,即得0.05 mg•mL-1的对照品溶液。
2.1.2 供试品溶液的制备 精确称取一点红的根、茎、叶粉末各2g,分别置250mL棕色量瓶中,加水150mL,放置过夜,超声处理10min,放冷,用水稀释至刻度,静置,滤过,弃去初滤液50mL,精密量取续滤液20mL,置100mL棕色量瓶中,用水稀释至刻度,摇匀,即得。
2.1.3 标准曲线的制备 精密量取没食子酸对照品溶液1.0、2.0、3.0、4.0、5.0mL分别置25mL棕色量瓶中,各加入磷钼钨酸试液1mL,再分别加水11、10、9、8、7mL用29%碳酸钠溶液稀释至刻度,摇匀。以随行试剂为空白。将对照品溶液在400~900nm波长范围内进行光谱扫描,确定最大吸收波长为760nm。30min后,于760nm波长处测定吸光度。以吸光度为纵坐标,浓度为横坐标,绘制标准曲线。计算得回归方程为: Y=0.0909X +0.0519,r=0. 9999,结果表明,没食子酸浓度在2～10mg•L-1 的范围内,与吸光度有良好的线性关系。, 百拇医药(张 燕 张洪斌 陈光宙 王 燕)
关键词:一点红;鞣质;绿原酸;含量测定
中图分类号:R284.1 文献标识码:A 文章编号:1673-7717(2010)01-0125-03
Determination of Tannins and Chlorogenic Acid of Different Vegetative Organs in Emilia sonchifolia (Linn.)DC
ZHANG Yan, ZHANG Hongbin, CHEN Guangzhou,WANG Yan
(Department of Life and Science, Qiongzhou College, Wuzhishan 572200,Hainan, China)
Abstract:Objective: To establish the analytical methods for determinating the contents of tannins and chlorogenic acid of different vegetative organs in Emilia sonchifolia (Linn.)DC. Methods: The content of tannins, expressed as gallic acid equivalents, was measured by using the phospho molybdenum tungstic acid-casein colorimetry, and the content of chlorogenic acid, expressed as chlorogenic acid equivalents, was measured by HPLC. Results: The good linearity between concentrations and absorbances of gallic acid was found in the range of 2～10 mg L-1, the average recovery was 100.32% and RSD was 1.45%, the contents of tannins in leaves, stems and roots of Emilia sonchifolia (Linn.)DC were 0.25%,0.15% and 0.22% respectively; The good linearity between content and peak area integral value of chlorogenic acid was found in the range of 0.0356~0.5340μg, the average recovery was 99.37% and RSD was 1.52%, the contents of chlorogenic acid in leaves, stems and roots of Emilia sonchifolia (Linn.)DC were 1.28%,1.14% and 0.91% respectively. Conclusion: Two proposed methods are sensitive, precise, accurate and reproducible, and they are suitable for the determination of tannins and chlorogenic acid in Emilia sonchifolia (Linn.)DC.
Key words:Emilia sonchifolia (Linn.)DC; tannins; chlorogenic acid; determination
一点红[Emilia sonchifolia (Linn.)DC][1],为菊科一点红属植物,一年生或多年生草本,又名叶下红、羊蹄草、红背叶、山羊草、野芥兰、牛尾膝等,在我国南方各省均有分布。一点红全草入药[2],清热凉血,活血化瘀,利尿消肿,消炎解毒。主治上呼吸道感染、咽喉肿痛、口腔溃疡、肺炎、急性肠炎、细菌性痢疾、泌尿系统感染、睾丸炎、乳腺炎、疖肿疮疡、皮肤湿疹、跌打扭伤。它还是目前市场上畅销的妇科良药——花红冲剂的主要原料。近些年来,一点红又成为人们喜爱的一种野菜[3],食味柔滑,口感清爽。
目前对一点红的研究主要集中在部分化学成分、栽培技术等方面[4-7],对其次生代谢产物含量的研究较少。本文对一点红不同营养器官中鞣质和绿原酸含量进行了测定分析,为一点红的药用质量控制及其次生代谢产物的进一步研究和开发提供参考依据。
1 材料与试剂
1.1 材料
一点红采自海南五指山市郊(由本系林教授鉴定为一点红全草),将其洗净,分成根、茎、叶三部分,在40℃条件下烘干至恒重,粉碎,过40目筛,放在干燥器中备用。
1.2 主要仪器LC-2010c型高效液相色谱仪(LCSOLUTION工作站,日本岛津公司);BP211D电子天平(德国Sartorius公司);UV-2100双光束紫外可见分光光度计(北京瑞利分析仪器公司);CO250型超声波清洗器(上海超声仪器厂)。
1.3 试剂没食子酸对照品(中国药品生物制品检定所,批号110831-200302);绿原酸对照品(中国药品生物制品检定所,批号110753-200413);磷钼钨酸试液(按《中华人民共和国药典》2005年版一部附录ⅩⅤB配制)[8];乙腈为色谱纯;水为二次蒸馏水;其他试剂均为分析纯。
2 方法与结果
2.1 鞣质含量测定
2.1.1 对照品溶液的制备精密称取没食子酸对照品50mg,置100mL棕色量瓶中,加水溶解并稀释至刻度,精密量取5mL,置50mL棕色量瓶中,用水稀释至刻度,摇匀,即得0.05 mg•mL-1的对照品溶液。
2.1.2 供试品溶液的制备 精确称取一点红的根、茎、叶粉末各2g,分别置250mL棕色量瓶中,加水150mL,放置过夜,超声处理10min,放冷,用水稀释至刻度,静置,滤过,弃去初滤液50mL,精密量取续滤液20mL,置100mL棕色量瓶中,用水稀释至刻度,摇匀,即得。
2.1.3 标准曲线的制备 精密量取没食子酸对照品溶液1.0、2.0、3.0、4.0、5.0mL分别置25mL棕色量瓶中,各加入磷钼钨酸试液1mL,再分别加水11、10、9、8、7mL用29%碳酸钠溶液稀释至刻度,摇匀。以随行试剂为空白。将对照品溶液在400~900nm波长范围内进行光谱扫描,确定最大吸收波长为760nm。30min后,于760nm波长处测定吸光度。以吸光度为纵坐标,浓度为横坐标,绘制标准曲线。计算得回归方程为: Y=0.0909X +0.0519,r=0. 9999,结果表明,没食子酸浓度在2～10mg•L-1 的范围内,与吸光度有良好的线性关系。, 百拇医药(张 燕 张洪斌 陈光宙 王 燕)