HPLC法同时测定香加皮中杠柳毒苷和4-甲氧基水杨醛的方法改进

[目的]建立同时测定香加皮中杠柳毒苷和4-甲氧基水杨醛含量的方法。[方法]采用高效液相色谱(HPLC)法,ODS色谱柱,流动相为乙腈∶甲醇∶水=27∶10∶63,检测波长220nm。[结果]杠柳毒苷线性范围0.08～1.0μg,r=0.9993;4-甲氧基水杨醛线性范围0.22～3.0μg,r=0.9999;杠柳毒苷和4-甲氧基水杨醛精密度的峰面积(RSD)值分别为0.79%、0.67%,重现性的RSD值分别为1.2%、2.2%,加样回收率分别为97.1%、96.2%。[结论]本法可同时测定香加皮中杠柳毒苷和4-甲氧基水杨醛的含量,分析时间短,且改进方法的准确度和重现性要优于文献报道方法。     

HPLC法同时测定香加皮中杠柳毒苷和4-甲氧基水杨醛

[目的]同时测定香加皮中杠柳毒苷和4-甲氧基水杨醛的含量。[方法]采用HPLC法,ODS色谱柱,流动相为乙腈∶水=27∶73,检测波长220nm。[结果]杠柳毒苷线性范围0.05～1.6μg,r=0.9994;4-甲氧基水杨醛线性范围0.23～7.4μg,r=0.9993;杠柳毒苷和4-甲氧基水杨醛的精密度分别为1.2%、3.0%,重现性分别为1.4%、2.8%,加样回收率分别为100.2%、99.8%。[结论]本法简便、准确,可同时测定香加皮中的杠柳毒苷和4-甲氧基水杨醛。     

香加皮产生毒副反应的原因及安全用药对策

从药学角度对香加皮产生毒副反应的原因进行分析:原因为香加皮饮片中强心苷成分杠柳毒苷含量差异大;与药材的新鲜程度和用药部位有关;与香加皮不同的用药方式有关.对香加皮安全用药提出对策为:完善香加皮饮片的质量标准,增加饮片中杠柳毒苷的含量测定;改变香加皮临床用药方式,以具有明确杠柳毒苷含量的提取物或中药配方颗粒的形式调配;在中成药中以香加皮提取物的形式入药,投料量以杠柳毒苷量计算;杠柳毒苷与人参皂苷配伍以达到减毒作用.     

香加皮有效部位滴丸的质量标准研究

[目的]建立香加皮有效部位滴丸的质量标准。[方法]建立了高效液相色谱(HPLC)法进行杠柳毒苷含量测定和杠柳毒苷的均匀度检查;对滴丸剂进行了常温下的初步稳定性考察。[结果]香加皮有效部位滴丸含量测定方法学考察的指标均符合有关规定;滴丸均匀度符合要求。[结论]建立了有效部位及滴丸的质量标准,方法简便、可靠。滴丸剂质量基本稳定。     

天津市区零售药店五加皮用药情况调查

[目的]调查天津市区零售药店中五加皮的用药情况。[方法]以香加皮中的杠柳毒苷为指标成分,采用高效液相色谱(HPLC)以及薄层层析色谱(TLC)法进行含量测定和定性鉴别。[结果]在32个样品中,31个样品为香加皮,1个为五加皮。[结论]天津市区零售药店中存在以香加皮代五加皮的现象。     

HPLC法测定不同产地香加皮中杠柳毒苷的含量

目的:测定不同产地香加皮中杠柳毒苷的含量。方法:采用反相高效液相色谱法,以乙腈-水(26∶74)为流动相,检测波长为220 nm。结果:17个产地的香加皮中杠柳毒苷的含量在0.006%~1.215%之间,其中以天津蓟县最高,山东泰安最低。结论:不同产地香加皮中杠柳毒苷的含量差异较大。     

三七总皂苷对杠柳毒苷药代动力学的影响

[目的]建立液相色谱-质谱联用(LC-MS/MS)定量分析方法,考察三七总皂苷对杠柳毒苷在大鼠体内的药动学影响。[方法]将18只大鼠随机分成杠柳毒苷单独给药组、杠柳毒苷和低剂量三七总皂苷配伍组、杠柳毒苷和高剂量三七总皂苷配伍组,大鼠连续口服给药7 d,在最后1次给药前和给药后不同的时间点从眼底静脉丛取血,采用LC-MS/MS法,以补骨脂素为内标,测定大鼠体内杠柳毒苷的含量,并运用DAS 1.0软件计算待测物的药动学参数。[结果]杠柳毒苷在1~500 ng/mL的范围内线性关系良好,方法学符合要求。药动学结果表明,杠柳毒苷配伍三七总皂苷后,杠柳毒苷在大鼠血浆中的主要药动学参数C_(max)、AUC_(0-t)、AUC_(0-∞)与杠柳毒苷单独给药相比存在统计学差异。[结论]本研究所建立的LC-MS/MS方法快速、灵敏、准确,可用于杠柳毒苷在大鼠体内的药动学研究。三七总皂苷对杠柳毒苷在大鼠体内的药动学存在影响。     

UPLC同时测定香加皮中六种成分

目的:了解不同产地、不同批次香加皮药材中6种成分的含量差异,评价不同产地、不同批次香加皮的质量。方法:采用超高效液相色谱法(UPLC)对香加皮中6种成分进行含量测定,比较不同产地间6种化合物的含量差异。结果:经方法学验证,6种化合物在线性范围内线性关系良好(r0.999 6),平均回收率在99.0%～104%(RSD4.47%)。发现香加皮中4-甲氧基水杨醛的含量均符合《中华人民共和国药典》规定要求,其中不同批次香加皮药材中杠柳毒苷、杠柳苷元和杠柳次苷含量差异较大。结论:所建立的超高效液相色谱法能够同时准确测定香加皮药材中的6种化学成分,并对香加皮中多种成分进行定量,为其质量评价提供了新方法。     

HPLC法测定香加皮中杠柳毒苷的含量

香加皮Perploca sepium Bge,又名北五加皮、杠柳皮，功效为祛风湿、强筋骨，主要用于风寒湿痹、腰膝酸软、心悸气短、下肢浮肿等，近年来常用于慢性充血性心力衰竭的治疗。香加皮的主要化学成分为10余种杠柳苷，其中杠柳毒苷又名杠柳苷G、萝蘼毒苷(periplocin，periploside G)为强心苷成     

香加皮强心成分杠柳毒苷肠菌代谢研究

[目的]检测杠柳毒苷在大鼠和人体肠菌作用下含量与成分的变化。[方法]采用离体粪便温孵法,利用高效液相—紫外法对温孵不同时间的杠柳毒苷及其代谢产物含量进行测定,并利用高效液相—质谱联用法对代谢物进行鉴定。[结果]杠柳毒苷含量随温孵时间的延长呈快速下降的趋势,温孵5h含量降低约75%,与此同时有代谢物生成,并随温孵时间的延长呈上升趋势。[结论]杠柳毒苷在肠菌(大鼠/人体)作用下,代谢迅速,生成了代谢产物杠柳次苷。     

Antagonism of Cortex Periplocae extract-induced catecholamines secretion by Panax notoginseng saponins in cultured bovine adrenal medullary cells by drug combinations

Ethnopharmacological relevance

Traditional Chinese Medicine (TCM) operates on the general principle that compatible components of different herbal decoction may work together to synergistically enhance therapeutic efficacy or reduce adverse effects. Cortex Periplocae is an herb that has been used in TCM clinics for a long time in the treatment of chronic heart failure. However, recently, the use of this herb has been restricted because of widespread abuse and misapplications. Radix Notoginseng is another herb that is used in TCM because of its protective role on cardiomyocytes. From our previous studies on these two herbs in a mouse model, we observed an increased LD₅₀ after oral administration of Cortex Periplocae extract (CPE) and Panax notoginseng saponins (PNS) in a ratio of 1:1 compared with Cortex Periplocae extract used alone.

Aim of the study

This study aimed to investigate whether there are mutual synergistic effects of the two herbal extracts, CPE and PNS, on catecholamines (CAs) secretion, and their possible underlying mechanism(s) for such effects.

Materials and methods

CPE and PNS were quantified by the LC-MS/MS method. HPLC-ECD was used to determine the CAs secreted into the medium by bovine adrenal medulla cells (BAMCs) and calcium influx was measured using a Calcium 4 reagent kit.

Results

We found that the stimulatory effect of CPE on CAs secretion was inhibited when used together with PNS. For a better clarification of the different constituents of the extracts, a quantitative analysis was carried out. Periplocin was found to be the main active component of CPE valued as 0.99% and saponins were the principal constituents of PNS. These results also showed that CPE increased the secretion of CAs in a dose-dependent manner while the actions of PNS were seen to be inhibitory. Periplocin monomer of CPE could be implicated for the actions of CPE since it plays the role of increasing the ACh-induced CAs secretion in a calcium-dependent manner. We therefore conclude that; CPE and PNS exert antagonistic effects in regulating the concentration of intracellular calcium.

Conclusions

PNS inhibits CPE-induced CAs secretion by suppressing calcium influx in bovine adrenal medulla cells while periplocin, one of the main components of CPE has the same secretagogue effect as CPE.     

建立评价香加皮质量的RP-HPLC色谱指纹图谱分析方法

目的:建立评价香加皮质量优劣的指纹图谱分析方法。方法:以香加皮药材为研究对象,应用RP-HPLC法,色谱柱为Krom asil C18(250 mm×4.6 mm,5μm),乙腈-0.025%磷酸水二元梯度洗脱模式,流速为1.0 m l/m in,检测波长为220 nm,建立了15批不同产地香加皮样品的RP-HPLC色谱指纹图谱。结果:对各产地药材指纹图谱进行聚类分析,根据聚类分析相似度分析结果,将香加皮药材分为4类,选定其中6批优质样品建立共有模式,并应用“中药色谱指纹图谱相似度评价系统2004版”对各产地药材进行了质量评价。结论:该方法简便、可靠,为有效控制香加皮药材质量提供依据。     

多成分定量结合指纹图谱分析用于不同产地香加皮的质量评价

采用超高效液相色谱法(UPLC),以甲醇-0.1％磷酸水溶液梯度洗脱,对山西、河南两省共30批香加皮中的3种成分(绿原酸、4-甲氧基水杨醛、杠柳毒苷)进行同时测定,并对其指纹图谱进行研究.结果表明不同产地样品中,3种成分含量差异较大;香加皮药材指纹图谱共标定17个共有峰,除H3,H11,样品相似度均在0.90以上.以3种成分含量及相似度数据进行显著性差异分析、聚类分析及主成分分析,结果表明不同产地的香加皮药材质量存在一定差异.建立的含量测定结合指纹图谱分析方法可更好地用于香加皮的质量评价.     

地榆四黄散的提取工艺优化及凝胶剂的制备

[目的]优选地榆四黄散的提取工艺,并制备凝胶剂。[方法]采用正交实验法,以没食子酸、黄芩苷、黄芩素和大黄类相关成分的提取含量为指标,进行方差分析,筛选最优提取工艺,并制备凝胶剂。[结果]四黄最佳提取工艺为用10倍量的50%乙醇热回流提取3次,每次1.5 h;地榆为用8倍量的水,回流提取3次,每次提取1.5 h。用1.5%的卡波姆及中药提取液制备凝胶剂的工艺可行。[结论]该凝胶剂制备工艺简便,质地细腻均匀。     

正交试验法优选胃安冲剂的制备工艺

目的；优选胃安冲剂的制备工艺，方法：以厚朴酚，白术多糖，盐酸小檗碱为指标成分，采用正交试验法对胃安冲剂的制备工艺进行优选，结果：优化工艺为：厚朴，枳实采用水提醇沉（20倍量水，0．5h，提取3次，95％乙醇沉淀杂质），黄连采用95％乙醇回流提取，白术等其它药材采用水提法（24倍量水，提取2次，每次2h，白术粒度为2－4mm(，结论：该工艺合理，有效成分提取效率高。     

清宫消炎栓中黄柏提取工艺优选

目的:用正交实验优选清宫消炎栓中黄柏乙醇回流提取的最佳工艺条件.方法:采用正交设计法,以出膏率、盐酸小檗碱含量为考察指标,对清宫消炎栓中黄柏回流的乙醇浓度、提取次数、乙醇倍量三因素进行优选研究.结果:乙醇浓度为70%,提取次数3次,乙醇倍量为5倍生药量,提取时间为1.5h.结论:乙醇回流浓度、提取次数是影响黄柏中盐酸小檗碱提取的主要因素,该工艺方法简单,适合工业化生产.     

秦皮总香豆素超声辅助提取工艺优化

目的:优化秦皮总香豆素的超声醇提工艺。方法:采用单因素试验方法研究超声辅助提取过程中乙醇体积分数、料液比、提取温度、提取时间和超声频率对秦皮总香豆素提取率的影响,并采用正交试验确定最佳提取工艺条件。结果:秦皮总香豆素超声辅助提取的最佳提取工艺为:按1∶10(质量-体积比)的料液比加入60%乙醇,于50℃条件下用175 W超声辅助提取50 min,秦皮总香豆素提取率可达到6.283%。结论:与传统索氏提取相比,该工艺具有耗时少、耗能少和提取率高等优点。     

Improvement of Growth and Periplocin Yield of Periploca sepium Adventitious Root Cultures by Altering Nitrogen Source Supply

Objective To increase the ultimate yield of periplocin in Periploca sepium adventitious root cultures by a two-stage culture based on nitrogen source.Methods Firstly,the effects of nitrogen source(NH4+-NO3-) at different ratios and different total initial nitrogen amounts on the accumulation of biomass and secondary metabolites in adventitious root cultures of P.sepium were investigated,and growth and production media for the two-stage culture based on the above results were established.Results The highest biomass and periplocin content were obtained in the culture medium of 15 mmol/L total nitrogen amount with NH4+-NO3-(1:2) and 30 mmol/L total nitrogen amount with nitrate as the sole nitrogen source.By adopting a fed-batch cultivation strategy,the dry weight adventitious root,periplocin content and yield were increased by 136%,108%,and 389%,respectively when compared with those of the control,reaching up to 8.13 g/L,157.15 μg/g,and 1277.63 μg/L,respectively.Furthermore,it was found that in the process of two-stage culture,the adventitious roots grew thicker significantly after they were transferred into production medium directly.Conclusion The ultimate yield of periplocin in P.sepium adventitious root cultures could be significantly increased by a two-stage culture based on nitrogen source.