中药胃肠安丸给药后木香烃内酯及去氢木香内酯药代动力学研究

采用HPLC-MS/MS多反应监测(MRM)定量分析,以地西泮为内标物在正离子模式条件下建立血样中木香烃内酯(Co)及去氢木香内酯(De)药物浓度的测定方法,探讨胃肠安丸口服给药后倍半萜内酯类成分Co及De的药代动力学过程,为进一步揭示胃肠安丸抗腹泻作用的物质基础提供理论依据。血浆样品中Co在0.700 0~769.7μg·L-1,De在2.510~956.0μg·L-1线性关系良好;且精密度、稳定性及提取回收率实验结果表明该类成分血药浓度的测定方法稳定可靠。经测定口服给药后Co及De血药浓度均随给药剂量的增加而增大,Tmax在10.65~12.98 h,表明倍半萜内酯类成分血药浓度达峰值时间较晚,Co及De Cmax分别为3.750~5.450,15.34~44.52μg·L-1;Co及De在体内吸收符合一室开放模型,其体内血药浓度达峰值所需时间较长,为揭示胃肠安丸作用物质基础及指导临床用药提供实验依据。     

提取木香中木香烃内酯及去氢木香内酯影响因素的初步研究

目的 考察溶媒、温度、加热时间对木香活性成分木香烃内酯和去氢木香内酯的影响.方法 用水和乙醇回流提取木香,分别将其提取物控制在不同的温度和时间范围内加热,用HPLC法测定木香烃内酯和去氢木香内酯的含量变化.结果 醇提取物中木香烃内酯和去氢木香内酯的含量远远高于水提取物,以乙醇为溶媒,两成分含量变化小;以水为溶媒,随着温度升高,加热时间延长,两成分下降明显,尤其是木香烃内酯.结论 溶媒、温度对木香烃内酯影响最为显著,时间因素次之,在含有木香的制剂工艺制备过程中要充分考虑这些影响因素,最大程度地保留有效成分.     

木香中去氢木香内酯和木香烃内酯提取工艺优选

目的:优选木香中去氢木香内酯和木香烃内酯的提取工艺。方法:以木香烃内酯和去氢木香内酯提取率为指标,采用单因素试验考察提取方法;以木香烃内酯和去氢木香内酯提取率为指标,选取乙醇体积分数、料液比、提取时间及提取次数为考察因素,通过正交试验优选2种内酯的提取工艺。结果:最佳提取工艺为采用乙醇温浸法,加6倍量90%乙醇于40℃温浸提取2次,每次2 h。木香烃内酯提取率92.3%,去氢木香内酯提取率94.1%,得膏率12.7%。结论:优选的提取工艺稳定可行,木香中2种内酯的提取率高,适用于工业大生产。     

不同产地和采收时间木香药材中木香烃内酯和去氢木香内酯的测定

目的:测定不同产地和采收时间木香药材中木香烃内酯和去氢木香内酯的含量.方法:采用高效液相色谱法,色谱柱为Venusil XBP C18(4.6 mm×250 mm,5μrn),以甲醇-水(65:35)为流动相,检测波长225 nm,流速1.0 mL· min-1,柱温30℃.结果:木香烃内酯线性范围为0.2～3.0 μg(r =0.9997),去氢木香内酯线性范围为0.2～3.0 μg(r =0.9998),木香烃内酯、去氢木香内酯回收率分别为99.27％,99.46％.结论:不同产地木香药材在9～12月期间采收均符合药典标准,10月份云南采收木香质量最佳.     

微波辅助提取木香有效成分的研究

目的:微波提取优选木香有效成分的最佳工艺条件。方法:采用正交试验法,通过高效液相法测定其指标成分木香烃内酯和去氢木香烃内酯,对微波提取木香有效成分的各影响因素进行考察。结果:优选木香提取工艺为微波功率400 W,以60%乙醇4倍量提取4次,每次20 min。结论:表明微波的提取时间短,能耗低,提取效率高,优于常规回流提取方法。     

木香药材中木香烃内酯和去氢木香内酯的定性和定量研究

目的 :对木香药材中两种倍半萜内酯进行定性定量研究。方法 :采用薄层色谱法和高效液相色谱法。薄层采用硅胶G板 ,展开剂为氯仿 -环己烷 (5∶ 1) ,1%的香草醛硫酸溶液 10 5℃加热显色。高效液相采用 Agilent Zorbax SB- C1 8色谱柱(4 .6 mm× 15 0 mm,5μm) ,甲醇 -水 (6 5∶ 35 ) ,检测波长 2 2 5 nm。结果 :两种倍半萜内酯分离良好。结论 :该方法可从定性定量两方面控制木香药材的质量。     

六味木香散中木香烃内酯去氢木香内酯的含量测定

目的:建立六味木香散中木香的主要有效成分木香烃内酯、去氢木香内酯的含量测定方法。方法:采用反相高效液相色谱法,Allteah C_(18)色谱柱(5μm,250mm×4.6mm),流动相为甲醇-水(65:35),流速为1.0ml/min,检测波长为225nm。结果:木香烃内酯线性范围为119.48ng～1792.2ng(r=0.999)、去氢木香内酯线性范围为122.84ng～1842.6ng(r=0.999),平均加样回收率为100.3%、99.8%。结论:该方法灵敏、准确、重复性好,具有专属性。     

“一测多评”测定木香中木香烃内酯和去氢木香内酯的含量

正木香为菊科植物木香Aucklandia lappa Decne.的干燥根,其中木香烃内酯(costunolide)和去氢木香内酯(dehydrocostuslactone)为挥发油的主要成分,此外还有菊糖和木香碱等。木香主要起行气调经止痛功效,是治脾胃虚寒气滞要药。木香烃内酯和去氢木香内酯均为倍半萜内酯,性质不稳定,不易保存     

HPLC法测定六味木香胶囊中木香烃内酯和去氢木香内酯

目的建立以HPLC法测定六味木香胶囊中木香烃内酯和去氢木香内酯的方法。方法色谱柱为VP-ODS柱(250mm×4.6 mm,5μm),流动相为甲醇-水(72∶28),检测波长为225 nm,柱温为25℃,体积流量为1.0 mL/min。结果木香烃内酯、去氢木香内酯进样量分别在203.12~2 031.20、207.84~2 078.40 ng与峰面积积分值呈良好的线性关系(r=0.999 9);平均回收率分别为101.1%、101.2%,RSD分别为0.48%、0.73%(n=6)。结论建立的方法简便、准确、重复性好、专属性强,可用于该制剂的质量控制和评价。     

HPLC测定木香槟榔丸中 木香烃内酯和去氢木香内酯的含量

目的:建立木香槟榔丸中木香烃内酯和去氢木香内酯的含量测定方法。方法:采用HPLC,色谱柱为Agilent C18(4.6 mm×250 mm,5μm),流动相甲醇-水(65∶35),检测波长225 nm,柱温30℃,流速1.0 mL.min-1。结果:木香烃内酯和去氢木香内酯的线性范围分别为0.046 4～0.3 712μg(r=0.999 6)和0.045 2～0.361 6μg(r=0.999 5);平均加样回收率分别为99.0%(RSD 0.86%,n=6)和99.2%(RSD 0.23%,n=6)。结论:该法简便,快速,结果准确,重复性好,可用于控制木香槟榔丸的质量。     

Pharmacokinetic study on costunolide and dehydrocostuslactone after oral administration of traditional medicine Aucklandia lappa Decne. by LC/MS/MS

Ethnopharmacological relevance

Radix Aucklandiae (RA), a well known traditional Chinese medicine, is widely used for treating various problems in digestive system. A selective and sensitive high-performance liquid chromatography coupled with mass spectrometry method was first developed and validated for simultaneous quantification of costunolide and dehydrocostuslactone in rat plasma with diazepam as internal standard after oral administration of RA extraction.

Materials and methods

Plasma samples were extracted via solid-phase extraction and detected by multiple-reaction monitoring mode under positive electrospray. Chromatographic separation was accomplished on an Agilent C₁₈ column (2.1 mm×150 mm, 5 µm), with 0.1% formic acid and acetonitrile (1:1) as the mobile phase at a flow rate of 0.5 mL/min.

Results

The quantification was performed using the transitions of m/z 233/187 for costunolide, m/z 231/185 for dehydrocostuslactone and m/z 285/193 for diazepam, respectively. Calibration curves were linear over the concentration range of 0.7–769.7 ng/mL for costunolide and 0.9–956.0 ng/mL for dehydrocostuslactone. The intra-day and inter-day precisions (RSD%) for two compounds was less than 8.76% and 9.70% and the accuracy (RE%) range from 6.14% to 5.35%. The time to reach the maximum plasma concentration (T_max) was 10.46 h for costunolide, 12.39 h dehydrocostuslactone. The elimination half-time (t_1/2) of costunolide and dehydrocostuslactone was 5.54±0.81 and 4.32±0.71 (h). The AUC of costunolide and dehydrocostuslactone was 308.83 and 7884.51 respectively (ng h/mL).

Conclusions

It was the first report for the study of pharmacokinetic profile of costunolide and dehydrocostuslactone in rat plasma after oral administration of RA extract. These results provided a meaningful basis for better understanding the absorption of traditional medicine, RA, and provide useful scientific data for clinical application.     

Effect of co-existent components in CO2 supercritical fluid extract of Angelica Sinensis Radix on metabolism of Z-ligustilide after oral administration in rats

ObjectiveTo establish a basis for Angelica Sinensis Radix (ASR) as a dietary supplement for colorectal cancer chemoprevention, the effect of co-existent components in supercritical fluid extract (SFE) of ASR on the pharmacokinetics of Z-ligustilide after oral administration was investigated in vitro and in vivo.MethodsIncubation in gastrointestinal contents and incubation in rat liver tissue homogenates post-mitochondrial supernatant (PMS) experiments were used to study changes in the levels of Z-ligustilide in vitro.ResultsWithin 4 hours, the level of Z-ligustilide in SFE declined at a slower rate than in its pure form. Clearance of Z-ligustilide after administration in its pure form was significantly slower than that of SFE of ASR (CL, 0.96 ± 0.16 mL·min/kg versus 1.24 ± 0.21 mL·min/kg P < 0.05; AUC, 243.37 ± 16.84 versus 176.69 ± 12.59 mg·min/L).ConclusionThese phenomena may be attributed to the interactions between the co-existent components in SFE of ASR and Z-ligustilide enhancing the stability of Z-ligustilide. These results suggest that the bioavailability of Z-ligustilide in SFE of ASR is improved. However, stabilization of plasma concentration was not sustained, so that the efficacy of active components could not be maintained. Thus, further processing of SFE of ASR is required.     

HPLC法同时测定香连片中两种内酯

目的 建立高效液相色谱法测定香连片中木香烃内酯和去氢木香内酯.方法 采用高效液相色谱法,色谱柱为kromasil-C18(4.6 mm ×250 mm,5μm),甲醇-水为流动相梯度洗脱,体积流量1.0mL/min,检测波长225 nm,柱温30 ℃.结果 木香烃内酯和去氢木香内酯线性质量浓度范围分别为0.7584～3.792 μg(r=0.9999),0.7488～3.744 μg (r=0.9999);平均回收率分别为95.98％(RSD为1.62％,n=6),95.04％(RSD为2.66％,n=6).结论 本法简单、准确、专属性强,可作为香连片的质量控制方法之一.     

Pharmacokinetics and bioavailability of cimicifugosides after oral administration of Cimicifuga foetida L. extract to rats

Ethnopharmacological relevance

Cimicifuga foetida L., a traditional Chinese medicine, has been used as an anti-inflammatory, antipyretic and analgesic remedy. The primary active constituents are believed to be present in the triterpene glycoside fraction.

Materials and methods

To develop an LC–MS/MS assay for four major cimicifugosides [cimicifugoside H-1 (Cim A), 23-epi-26-deoxyactein (Cim B), cimigenolxyloside (Cim C) and 25-O-acetylcimigenoside (Cim D)] obtained from C. foetida L. and apply it to investigate their pharmacokinetic (PK) properties and bioavailabilities through oral administration of C. foetida L. extract (12.5, 25 and 50 mg/kg) and single intravenous (i.v.) doses (5 mg/kg) of the individual cimicifugosides in rat. PK parameters were estimated by non-compartmental analysis.

Results

All calibration curves showed excellent linear regressions (all r>0.995) within the range of tested concentrations. The intra- and inter-day variations were <15% in terms of RSD. The molar ratio of Cims A, B, C, and D in the extract was 20.7:1.4:2.9:1. PK parameters for Cims A, B, C, and D following oral administration of the extract were respectively: C_max 4.05–17.69, 90.93–395.7, 407.1–1180 and 21.56–45.09 pmol/mL; T_max 0.46–1.28, 2.00–4.67, 14.67–19.67 and 8.08–14.27 h; absolute oral bioavailability (F) 1.86–6.97%, 26.8–48.5%, 238–319% and 32.9–48%. PK parameters after i.v. administration of individual cimicifugosides were respectively: elimination half-life 1.1, 2.5, 5.7 and 4.2 h; clearance 15.7, 0.48, 0.24 and 1.13 mL/h kg.

Conclusions

Systemic exposure to Cims B, C and D following oral administration of the extract was significantly greater than to Cim A despite the predominance of Cim A in the extract. Significantly different clearance and interconversion from Cim A to Cim C probably accounts for the different exposure to the four cimicifugosides.     

HPLC法同时测定藏药七味铁屑丸中木香烃内酯和去氢木香内酯

目的 建立以反相高效液相色谱法同时测定七味铁屑丸(铁屑、寒水石、藏木香、木香、甘青青蓝、红花,五灵脂膏)中木香烃内酯和去氢木香内酯的方法.方法 采用HPLC法,色谱柱为Welchrom Materials柱C18(250 mm ×4.6 mm,5μm),流动相甲醇-水(65∶35,v/v);检测波长225 nm;柱温30℃;体积流量1.0 mL/min.结果 木香烃内酯线性范围为0.011 5～0.115 mg/mL;去氢木香内酯线性范围为0.011 9～0.119 mg/mL,相关系数分别是0.9999和0.9998;平均加样回收率木香烃内酯为100.96％(RSD为1.28％,n=6);去氢木香内酯为100.74％(RSD为1.04％,n=6).结论 本法简便、准确、快速,可用于七味铁屑丸的质量控制.     

舒心片薄层色谱鉴别及木香烃内酯和去木香烃内酯的含量测定

目的制定舒心片质量标准方案,控制制剂成品质量。方法采用薄层色谱法对舒心片中的沉香、莪术等进行鉴别。采用HPLC法测定木香中木香烃内酯和去氢木香烃内酯总含量。色谱条件:以十八烷基硅烷键合硅胶为填充剂;乙腈-水(58∶42)为流动相;检测波长为225 nm。结果 TLC法对制剂中鉴别药材专属性强、斑点分离清晰、重现性好,阴性无干扰;HPLC测定木香烃内酯在0.023 7~0.237 0μg范围内呈良好的线性关系,去氢木香烃内酯在0.158 8~1.588 0μg范围内呈良好的线性关系,回收率为96.27%,RSD为1.17%。结论 TLC操作方便、分辨力高;HPLC法专属性强,重复性好,可作为舒心片制剂的质量控制标准。     

Comparative pharmacokinetics of baicalin after oral administration of pure baicalin, Radix scutellariae extract and Huang-Lian-Jie-Du-Tang to rats

Huang-Lian-Jie-Du-Tang (HLJDT) is an important “heat-clearing” multiherb remedy of traditional Chinese medicine, and Radix scutellariae (Scutellaria baicalensis Georgi, Labiatae) is a key ingredient herb in it. Baicalin and wogonoside are two main effective ingredients enriched in Radix scutellariae. In the present study, pharmacokinetic differences of baicalin following oral administration of pure baicalin, Radix scutellariae extract, baicalin co-administrated with extract of the other three herbs of HLJDT and HLJDT were investigated in male S.D. rats with approximately the same dose of 200 mg/kg baicalin. The pharmacokinetic comparison of wogonoside was conducted only in Radix scutellariae extract and HLJDT. Plasma concentrations of baicalin and wogonoside were determined using HPLC method. Unpaired Student's t-test was used for statistical comparison. The results indicated that baicalin and wogonoside demonstrated bimodal phenomenon in the plasma profile. Some ingredients in the other three herbs of HLJDT, not in Radix scutellariae itself, had pharmacokinetic interaction with baicalin and wogonoside and hence decreased their systematic exposure level (p < 0.01). The absorption site of baicalin was preliminary evaluated in rat using in situ absorption in stomach and different intestinal segments and results revealed the existence of double-site absorption of baicalin. The first absorption site was in upper intestinal, probably via directly absorption of baicalin; while the second absorption site was in colon in the form of aglygon.     

大豆异黄酮对盐酸雷洛昔芬在大鼠体内的药动学研究

目的:研究大豆异黄酮对盐酸雷洛昔芬在大鼠体内的药动学,并观察大豆异黄酮对盐酸雷洛昔芬药动学参数以及相对生物利用度Frel参数的影响。方法:取SD雌性大鼠8只(390±10)g,随机平均分为两组,对照组为盐酸雷洛昔芬片(RH)组,实验组为大豆异黄酮盐酸雷洛昔芬片(RH-SIF)组,按剂量10.7 mg/kg灌胃给药,分别于0.5、0.75、1、2、4、6、8、12、24、48、72、96、144、192 h取血,处理样品后,以RH为对照品,采用HPLC法测定两组血浆样品中的药物浓度并拟合药时曲线,得出药物动力学参数。采用AIC判别法确定两组制剂在大鼠体内的血药分布所符合的房室模型,并通过AUCT.DR/(AUCR.DT)计算得到RH-SIF片相对生物利用度Frel。结果:①以盐酸雷洛昔芬(RH)为检测指标,给药后两组大鼠体内的主要药动学参数分别为:实验组:Ka为(0.67±0.22)/h,Ke为(0.01±0.00)/h,Tmax为(2.13±0.12)h,Cmax为(0.78±0.02)μg/ml,AUC为(74.64±8.62)μg.h/ml;对照组:Ka为(0.27±0.02)/h,Ke为(0.01±0.00)/h,Tmax为(4.36±1.05)h,Cmax为(0.48±0.13)μg/ml,AUC为(62.73±5.53)μg.h/ml。②与市售的RH片相比,大鼠服用RH-SIF片后,达峰时间明显缩短,达峰浓度明显升高(P<0.05),且两制剂的药时曲线下面积AUC有显著差异(P<0.01)。③相对生物利用度Frel,ie:AUCT.DR/(AUCR.DT)=165.68%。结论:两组制剂在大鼠体内均符合权重为1/C2的单室模型分布,且大豆异黄酮盐酸雷洛昔芬片能明显提高雷洛昔芬的相对生物利用度。