HPLC法测定热毒清锭中延胡索乙素的含量

目的:建立高效液相色谱法,测定热毒清锭中延胡索乙素的含量.方法:色谱柱为Agilent HypersilODS柱(4.0×250mm,5μm),以甲醇-0.1%磷酸溶液(用三乙胺调节pH至6.0)(60:40)为流动相,检测波长282nm.结果:延胡索乙素在2～10μg范围内呈良好的线性关系(r=0.9999,n=5),平均回收率99.7%,RSD=2.78%(n=5).结论:本法简便,准确,灵敏度高.     

元胡止痛片中延胡索乙素的含量与镇痛作用相关性的研究

目的:测定市售不同厂家生产的元胡止痛片中延胡索乙素含量及其镇痛作用并作相关性的研究。方法:采用高效液相色谱法测定延胡索乙素的含量,通过小鼠扭体法进行镇痛作用实验。结果:元胡止痛片可以显著抑制醋酸引起的小鼠扭体反应并与其延胡索乙素的含量呈一定相关性。结论:可以用镇痛作用为指标代替延胡索乙素含量的测定更具合理性。     

延胡索及白芥子涂方中延胡索乙素经皮渗透的比较研究

目的比较研究延胡索及其复方白芥子涂方中延胡索乙素经皮渗透特性。方法采用Valia-Chien扩散池法进行体外经皮渗透实验,利用HPLC法测定接收液中延胡索乙素的含量。结果对Q-t线性回归,延胡索:Q=61.012t-33.779;R2=0.9907;Js=61.012μg/(cm2·h);白芥子涂方:Q=646.13t-2406.7,R2=0.9968;Js=61.012μg/(cm2·h)。白芥子涂方中延胡索乙素的Js是药材中延胡索乙素的10.59倍。结论复方中的细辛、白芥子、甘遂等成分对延胡索乙素经皮渗透有显著促进作用,本研究对白芥子涂方治疗哮喘提供理论基础。     

SLS-胶束荧光法测定元胡中延胡索乙素的含量

根据胶束具有增溶增敏的作用,在10余种表面活性剂中,筛选出十二烷基磺酸钠为胶束试剂,在优化的实验条件下,对元胡中镇痛作用最强的延胡索乙素进行了含量测定研究。用国产930型荧光光度计达到了高灵敏的检测,线性范围为2×10~(-9)～8×10~(-7) g/ml,最低检测限为2.5×10~(-11)g/ml,效果良好。     

Plasma pharmacokinetics and lung distribution of tetrahydropalmatine after topical application of cold asthma recipe extract: Feishu (BL 13) versus Non-Feishu acupoint

Ethnopharmacological relevance

Acupoint application of cold asthma recipe (CAR) was a traditional Chinese medicine (TCM) method, widely used as an alternative medicine for clinical prevention of the common winter diseases of asthma and bronchitis. Tetrahydropalmatine (THP) was a main active ingredient of CAR extract. The aim of this study is to compare plasma pharmacokinetics and lung distribution of THP between Feishu (FS) acupoint (BL 13) and Non-Feishu (NFS) acupoint application of CAR extract by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

Materials and methods

The extract of CAR was topically administrated in FS and NFS acupoint of rats for plasma pharmacokinetics, and topically administrated in FS and NFS acupoint of mice for lung distribution. The plasma and lung homogenates were pretreated by protein precipitation with acetonitrile. Chromatographic separation was performed on an ACQUITY UPLC BEH C18 column with a mobile phase consisted of 0.1% formic acid in acetonitrile and 0.1% formic acid in water. The detection was accomplished by multiple-reaction monitoring (MRM) scanning in the positive electrospray ionization (ESI⁺) mode. All pharmacokinetic parameters were estimated by non-compartmental analysis.

Results

A sensitive, accurate and precise UPLC-MS/MS method was successfully established for determination of THP in 100 μL plasma and lung homogenate. The lower limit of quantification (LLOQ) of THP was 0.05 ng/mL and 0.072 ng/mL, respectively. The pharmacokinetic results manifested that THP was absorbed and eliminated slowly in plasma. Additionally, it was found that there was significantly higher amount of THP absorbed into blood and lung after FS acupoint application compared to NFS acupoint application.

Conclusions

Both of the rat plasma pharmacokinetics and mice lung distribution of THP could support that FS acupoint application of CAR extract has greater advantages of absorption into the blood circulation and distribution in target tissue over NFS acupoint application. The results might be helpful in providing a rational explanation for why the TCM chose the acupoint application and elucidating the underlying mechanism of this treatment.     

Combination use of ferulic acid,ligustrazine and tetrahydropalmatine inhibits the growth of ectopic endometrial tissue: A multi-target therapy for endometriosis rats

Ethnopharmacological relevance

Ferulic acid (FA), ligustrazine (LZ) and tetrahydropalmatine (THP) are separately isolated from Chinese Angelica, Szechwan Lovage Rhizome and Rhizoma in the Jiawei-Foshou-San formula, a popular traditional Chinese medicine for irregular menses. It has been reported that the combination use of FA+LZ+THP has similar effect on endometriosis, but the underlying mechanisms are unclear. This study was to investigate the combination effects and mechanisms of FA+LZ+THP on endometriosis rats.

Materials and methods

Fifty endometriosis rats were intragastricly treated with FA+LZ+THP for 4 wk. The volume of ectopic endometrial tissue was measured to evaluate the effects. Then the changes in hypothalamic–pituitary–ovarian axis and ERE pathway were indicated by the levels of E₂, GnRH, FSH and LH, and the expressions of ER, HSP90 and COX-2, respectively. In addition, peritoneal macrophages of each rat were cultured in vitro and treated with (FA+LZ+THP)-medicated serum for 24 h. The proliferation and phagocytosis abilities, the levels of IL-1β and TNF-α, and the expression of IκBα were then measured for the changes of peritoneal macrophage activities.

Results

Combination use of FA+LZ+THP diminished the volume of the ectopic endometrial tissues (P<0.05 or P<0.01). It also decreased the E₂ level, suppressed the expression of GnRH, FSH and LH, and decreased the protein expression of ER, HSP90 and COX-2 (all P<0.05 or P<0.01). The phagocytosis ability of peritoneal macrophage was enhanced by (FA+LZ+THP)-medicated serum (P<0.05) with no change of proliferation (P>0.05). Moreover, IL-1β and TNF-α were downregulated (both P<0.05 or P<0.01) and IκBα was upregulated by the (FA+LZ+THP)-medicated serum (P<0.01).

Conclusions

The combination use of FA, LZ and THP could inhibit the growth of ectopic endometrial tissue in endometriosis rats. It might be related to the down-regulation of hypothalamic–pituitary–ovarian axis, the amelioration in ERE pathway and the improvement of peritoneal macrophage activities.     

夏天无提取物中原阿片碱和延胡索乙素的大鼠肠吸收特性

［摘要］目的考察夏天无提取物中有效成分原阿片碱和延胡索乙素的大鼠肠吸收特性。方法采用体外外翻肠囊模型研究原阿片碱和延胡索乙素的肠吸收特性。采用高效液相色谱（HPLC）法测定样品浓度,计算表观渗透系数（Papp）,并绘制累积吸收量-时间曲线图。考察并比较原阿片碱和延胡索乙素在不同肠段、不同给药浓度、不同pH条件下的Papp。结果原阿片碱和延胡索乙素在回肠段的Papp分别约为2.58×10 5和3.15×10 5 cm&#8226;s 1,与其他肠段比较,差异有统计学意义（P＜0.05）。原阿片碱在十二指肠、空肠和回肠的Papp随着给药体系浓度的升高而降低,累计吸收量呈非线性增加。给药体系pH由7.4降低到5.4时,原阿片碱和延胡索乙素在各肠段的吸收均呈降低的趋势。结论原阿片碱和延胡索乙素在回肠吸收较好,pH对于其在小肠内的吸收具有一定影响,原阿片碱的吸收在十二指肠、空肠和回肠可能存在饱和现象。     

补肾消石颗粒质量标准研究

目的:建立补肾消石颗粒的质量标准。方法:采用薄层色谱(TLC)法,对处方中的元胡、泽泻、山药进行定性鉴别;采用薄层色谱扫描法测定延胡索乙素的含量。结果:在TLC中均能检出元胡、泽泻、山药的特征斑点;延胡索乙素进样量在0.408～2.04μg范围内线性关系良好(r=0.9975),平均回收率为95.94%(RSD=1.85%)。结论:所建标准可用于该制剂的质量控制。     

高效液相色谱法同时测定元胡止痛片中欧前胡素、延胡索乙素与异欧前胡素的含量

目的：建立元胡止痛片中欧前胡素、延胡索乙素与异欧前胡素的HPLC含量测定方法。方法：采用Agilent ZORBAX SB-C18（4.6mm×250mm,5μm）色谱柱,流动相：甲醇-0.1%磷酸溶液（三乙胺调pH值至6.5）（61∶39）; 检测波长：280nm; 流速：1.0ml/min。结果：欧前胡素在0.0474～0.2844μg范围内与峰面积呈良好的线性关系（r=0.9999）; 延胡索乙素在0.0205～0.1230μg范围内与峰面积呈良好的线性关系（r=0.9998）,异欧前胡素在0.0750～0.4500μg范围内与峰面积呈良好的线性关系（r=1.0000）,平均回收率分别为98.6%、97.5%、96.8%。结论：本方法简便、准确、重现性好,可用于元胡止痛片的质量控制。     

四氢原小檗碱类药物对培养大鼠单个心肌细胞内游离Ca2+的影响

用Fura-2/AM技术和AR-CM-MIC阳离子测定系统,直接测定了四氢小檗碱(tetrahy-droberberine,THB)、左旋四氢巴马汀(l-tetrahydropalmatine,THP)和左旋千金藤立定(l-stepholidine,SPO)对培养大鼠单个心室肌细胞内游离钙([Ca²⁺]i)的影响,并与维拉帕米(Ver)做了比较。结果显示,THB,THP,SPD浓度为10～100μmol·L^-1时,均可使静息[Ca²⁺]i轻度升高,河豚毒素不能抑制之;浓度为1～100μmol·L^-1时,可明显抑制高K+引起的[Ca²⁺]i增高;30μmol·L^-1对胞外高Ca²⁺和去甲肾上腺素引起的[Ca²⁺]i增高也有明显的抑制作用,但均较Ver的抑制作用为弱;THPB对哇巴因引起的[ca²⁺]i升高无明显抑制作用。结果提示,THB,THP,SPD在抑制电压依赖性钙通道从而影响细胞膜[ca²⁺]i内流方面与Ver相似,但比Ver弱。