当归四逆汤中阿魏酸在大鼠血清中含量测定及药动学研究

目的:测定当归四逆汤(当归、白芍、桂枝、细辛、甘草、通草和大枣)口服后大鼠血清中阿魏酸血药浓度及药代动力学参数.方法:SD大鼠按20 g/kg灌胃给予当归四逆汤水煎液后,在7个时间点取血清,利用HPLC方法,通过外标法测定阿魏酸血药浓度,运用3P97软件计算药代动力学参数.结果:大鼠口服当归四逆汤水煎剂后阿魏酸药代动力学参数为Cmax 7.988 771 mg/L,AUC 7 861.507 80 mg/(L·min),Ke(1/min)0.001 222 669,Ka(1/min)0.019 541 92,t1/2(Ka)35.469 746 min,Tpeak 151.290 314 min.结论:大鼠口服当归四逆汤后阿魏酸吸收快,代谢比较慢.     

脉络宁注射液中肉桂酸血药浓度的LC-MS-MS测定及人体药动学研究

目的:建立LC-MS-MS法测定人血浆中肉桂酸的浓度,研究单剂量和多剂量静脉滴注脉络宁注射液后肉桂酸在健康人体内的药动学。方法:血浆样品以酸化的乙酸乙酯-乙醚(1∶3)萃取;ZOBAX SB C18色谱柱(4.6 mm×150 mm,5μm)分离,柱温为30℃,流动相为甲醇-水(2 mmol.L-1醋酸铵)(45∶55),流速为500μL.min-1;电喷雾离子源,多反应监测。检测离子:肉桂酸,[M-H]-,m/z146.8/103.1;替硝唑(内标),[M-H]-,m/z245.6/126.0。10名受试者单、多次剂量静脉滴注脉络宁注射液后,采用LC-MS-MS法测定血浆中肉桂酸浓度,DASver1.0软件对药-时曲线进行拟合,并计算药动学参数。结果:肉桂酸在0.50～400μg.L-1呈现良好线性,最低定量限为0.50μg.L-1,日内、日间精密度均小于10%。受试者给药后,肉桂酸药时曲线符合二房室模型。单、多次剂量主要药动学参数分别为Cmax(μg.L-1):115.73±44.31,113.79±25.61;T1/2β(h):0.41±0.087,0.52±0.132;V(L.kg-1):0.519±0.134,0.651±0.322;CL(L.kg-1.h-1):0.899±0.295,0.830±0.222;AUC0～t(μg.L-1.h-1):158.64±56.019,166.49±46.788。结论:该法准确、灵敏、专属。单剂量和多剂量给药后肉桂酸主要药动学参数经统计学分析无显著差异;多剂量给药后,肉桂酸体内无蓄积现象,体内过程不受性别差异的影响;其在受试者体内的药动学存在明显个体差异。     

不同制法四逆汤中甘草酸含量的比较

目的 对四逆汤传统汤荆、经方颗粒剂及单味配方颗粒剂中有效成分甘草酸的含量进行比较.方法 采用HPLc法测定3种不同剂型中甘草酸的含量.结果 甘草酸(每处方)在传统汤剂中的含量为92.92 mg,经方颗粒荆中的含量为66.95mg,单味配方颗粒中的含量为199.43mg.结论 3种剂型中指标性成分甘草酸的含量均有显著性差异.可为四逆汤剂型的进一步研究提供科学依据.     

3种方法制备的四逆汤中甘草苷、甘草酸含量测定

目的:对四逆汤传统法、药典收录的水提醇沉法、各单味药分提合并法制备的四逆汤中甘草苷、甘草酸含量进行比较.方法:采用HPLC测定3种不同的方法提取液中甘草苷、甘草酸的含量,C18色谱柱,流动相乙腈-0.05％磷酸水,梯度洗脱,柱温25℃,检测波长237 nm,流速1.0 mL·min -1,进样量10 μL.结果:甘草苷在传统汤剂中含量为2.57 mg·g-1,经方合剂中的含量为2.21 mg·g-1,单味配方合并液中含量为3.58 mg·g-1;甘草酸在传统汤剂中含量为1.96 mg·g-1,经方合剂中的含量为1.84 mg·g-1,单味配方合并液中含量为3.04 mg·g-1.结论:四逆汤单提合并法中甘草苷、甘草酸含量高,且各制法的四逆汤中甘草苷、甘草酸含量均有显著性差异,为四逆汤提取方法研究提供依据.     

四逆汤的药动学研究

目的：探讨四逆汤（附子，干姜，炙甘草，等）精制物药代动力学与药效动力学的相关性。揭示四逆汤的配伍规律。方法：应用同一来源含药血清，分别以乌头类生物碱和一氧化氮（NO）为指标，同步进行血药浓度测定和药理效应测定。结果：二者在犬体内均呈一级速度消除，具有开放一房室模型的特征，药动学参数K血与K效，TI/2血与TI/2效接近，均具有药效产生快，作用维持时间较长的特点，反映出四逆汤“走而不守，守而不走”的特性。结论：四逆汤的药代动力学与药效动力学参数相关性好。     

保心微丸中肉桂酸大鼠体内的药代动力学研究

目的研究保心微丸中肉桂酸在大鼠体内的药代动力学.方法采用高效液相色谱法测定肉桂酸血药浓度,色谱柱HypersilODSC18,150mm×4.6mm,5μm,柱温30C.流动相甲醇-1%醋酸(4555)流速0.5mL/min;检测波长273nm;进样量10μL.结果与结论建立了用高效液相色谱法测定肉桂酸血药浓度的方法,肉桂酸的峰面积(y)与浓度(X)之间的回归方程为y=4973.5348+42867.96678X,相关系数r=0.9998.肉桂酸在血浆中的回收率为97.5%,RSD为1.33%.检测限为0.15ng,大鼠血浆中最低检测浓度为75ng/mL.肉桂酸口服吸收在大鼠体内的药时过程为线性动力学过程,符合一级吸收一级消除的开放式室模型,t1/2(K.)为7.12min,tmax为53.29min,Cmax为0.20μg/mL,t1/2,2(Ke)为340.74min.该药代动力学参数可能是保心微丸中肉桂酸及有关成分等多种成分在大鼠体内的综合体现.     

中药复方苏冰口崩片中肉桂酸在大鼠体内的HPLC测定及药动学研究

目的：建立大鼠血浆中肉桂酸浓度的HPLC方法。方法：血浆样品经醋酸酸化,用氯仿提取药物。色谱柱Kromasil C18(4.6 mm×250 mm,5 μm),检测波长278 nm,柱温为室温,流动相为甲醇-乙腈-水-冰醋酸(25∶20∶55∶0.3),流速1.0 mL·min^-1,以卡马西平为内标,按内标法定量。结果：肉桂酸在4.0～400 ng·mL^-1线性关系良好(r=0.999 9),平均提取回收率为86.4%,方法回收率为100.3%。低、中、高3种浓度日内及日间RSD均小于6.0%。结论：所建立的RP-HPLC方法灵敏、专一、准确、精密,本方法为苏冰口崩片中肉桂酸的血药浓度监测及药动学研究提供了方法学基础。     

四逆汤中甘草酸的含量测定

目的探寻测定四逆汤甘草酸含量的方法.方法以甘草酸单铵盐为指标,采用薄层层析-紫外分光光度法测定其含量及回收率.结果甘草酸的含量为 0.5054mg/ml, RSD=4.07%,平均回收率为 97.14%.结论本方法快速、简便、准确,重现性好,可有效控制四逆汤的内在质量.     

山茱萸中马钱苷大鼠体内药动学研究

目的:建立高效液相色谱法研究山茱萸大鼠灌胃给药后马钱苷的体内药代动力学特点。方法:采用岛津LC-10ATvp高效液相色谱仪,浙江智达N2000工作站,Hypersil C18(4.6 mm×250 mm,5μm)色谱柱,流动相:乙腈-甲醇-0.1%磷酸溶液(10∶5∶85),流速1.0 mL·min-1,检测波长236 nm,检测山茱萸提取物大鼠灌胃后马钱苷的药代动力学参数。结果:马钱苷的药代动力学模型为单室模型一级吸收,其药代动力学参数为Ka=0.06 min-1,Ke=0.013 min-1,T(peak)=33.90 min,t1/2ka=12.47 min,t1/2ke=52.45 min。结论:高效液相色谱法研究山茱萸大鼠灌胃给药后马钱苷体内药代动力学,方法的专属性强,结果可靠。     

四逆汤合剂中甘草酸的含量测定

以甘草酸单铵盐为指标，采用薄层层析-紫外分光光度法定量，对四逆汤合剂中甘草酸的含量进行研究。结果甘草酸的含量为1．28mg／ml，n=5，RSD=4．45％，平均回收率为97．1％。实验结果表明，本方法快速、简便、准确，重现性好。     

小承气汤中橙皮苷和肉桂酸的含量测定

目的:建立小承气汤中橙皮苷和肉桂酸的HPLC同时测定方法,以便进行质量控制。方法:采用Apollo C18柱(250mm×4.6mm,5μm),流动相为乙腈和0.011%冰乙酸水溶液,梯度洗脱,时间40min,流速1mL/min,检测波长为283nm。结果:橙皮苷和肉桂酸的线性范围分别为0.20-0.80μg和0.05-0.33μg,精密度为0.24%和0.16%,平均回收率分别为96.3%和101.9%。在6批样品中,橙皮苷和肉桂酸的含量差别比较大。结论:文章建立的方法简便、灵敏、准确,可用于小承气汤的质量控制。     

高效液相色谱法测定疏冠流浸膏在大鼠体内阿魏酸的含量

 目的探讨口服疏冠流浸膏在大鼠血清中阿魏酸的测定方法。方法用RP-HPLC测定疏冠流浸膏中阿魏酸的含量,条件：C₁₈柱,以甲醇-1%醋酸水（35∶65）为流动相,紫外检测波长313nm。结果阿魏酸在40～200 μg·L^-1浓度范围内回归方程为Y=5.15×10^-6X+0.5824,相关系数r=0.9998。结论测定血清样品中阿魏酸用有机溶媒法较好。     

HPLC同时测定小承气汤中4个化学成分

目的:建立小承气汤中肉桂酸、大黄酸、芦荟大黄素和川陈皮素的HPLC同时测定方法。方法:采用Apollo C18柱(150mm×4.6mm,5μm),流动相为乙腈和0.011%冰乙酸溶液,梯度洗脱,时间25min,流速1mL/min,检测波长为254nm。结果:肉桂酸、大黄酸、芦荟大黄素和川陈皮素的线性范围分别为0.08-0.47、0.58-2.02、0.19-0.37和0.02-1.39μg,精密度为0.44%、0.17%、0.52%和0.33%,平均回收率分别为97.3%、99.1%、99.3%和100.5%。结论:建立的方法简便、灵敏、准确,可用于小承气汤的质量控制。     

Identification and pharmacokinetics of multiple constituents in rat plasma after oral administration of Yinchenzhufu decoction

Ethnopharmacological relevance

Yinchenzhufu decoction (YCZFD) is a classical Chinese herbal formula and has been used to treat severe jaundice in chronic liver injuries since the Qing Dynasty (18th century CE). To identify the components absorbed into the blood in YCZFD and explore their pharmacokinetic profile for understanding the effective ingredients of YCZFD.

Materials and methods

After rats were given YCZFD by intragastric administration, the plasma was processed by precipitation of protein. The compounds in YCZFD extract and the plasma were identified by using high-resolution mass spectrometry with a database-directed strategy. The pharmacokinetics of multiple compounds from YCZFD in rat plasma was studied by using the established UPLC-MS/MS method.

Results

Forty compounds in YCZFD extract and 21 prototype compounds with 11 metabolites in rat plasma were detected after oral administration. The pharmacokinetic parameters of glycyrrhizic acid, glycyrrhetic acid, cinnamic acid, ononin, atractylenolide III, and liquiritin from YCZFD were obtained in rats.

Conclusions

The identified constituents and the pharmacokinetic features of YCZFD are helpful for understanding the material bases of its therapeutic effects.     

Protective effects of cinnamic acid and cinnamic aldehyde on isoproterenol-induced acute myocardial ischemia in rats

Ethnopharmacological relevance

Cinnamomum cassia is a well-known traditional Chinese herb that is widely used for the treatment of ischemic heart disease (IHD). It has favorable effects, but its mechanism is not clear. To investigate the effects of cinnamic aldehyde (CA) and cinnamic acid (CD) isolated from Cinnamomum cassia against myocardial ischemia produced in rats by isoproterenol (ISO).

Materials and methods

Ninety male Sprague-Dawley rats were randomized equally to nine groups: a control group, an untreated model group, CA (22.5, 45, 90 mg/kg) or CD (37.5, 75, 150 mg/kg) treatment, or propranolol (30 mg/kg). Rats were treated for 14 days and then given ISO, 4 mg/kg for 2 consecutive days by subcutaneous injection. ST-segment elevation was measured after the last administration. Serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), nitric oxide (NO), and blood rheology were measured after the rats were sacrificed. The hearts were excised for determining heart weight index, microscopic examination, superoxide dismutase (SOD) and malondialdehyde (MDA) measurements.

Results

CA and CD decreased the ST elevation induced by acute myocardial ischemia, decreased serum levels of CK-MB, LDH, TNF-α and IL-6, and increased serum NO activity. CA and CD increased SOD activity and decreased MDA content in myocardial tissue.

Conclusion

CA and CD were cardioprotective in a rat model of ischemic myocardial injury. The protection was attributable to anti-oxidative and anti-inflammatory properties, as well as increased NO. The results support further study of CA and CD as potential treatments for ischemic heart disease.     

RP-HPLC测定盐酸青藤碱不同给药途径的血药浓度及药动学研究

 目的　采用RP HPLC测定盐酸青藤碱灌胃和皮肤给药后不同时间大鼠体内血清浓度 ,计算出灌胃和皮肤给药途径主要药动学参数。方法　采用RP HPLC以乙腈0.01mol·L^-1磷酸二氢钠溶液-四甲基二乙胺 (46∶54∶0.22)为流动相,磷酸调至pH6.9,C₁₈ODS柱(4.6mm×25.0mm,5μm),紫外检测波长为26.3nm。 结果　盐酸青藤碱在 0.25～10.0μg·mL^-1范围内呈良好线性关系(r=0.9995),方法的平均回收率达98.5%,RSD=13%。计算出灌胃给药途径主要药动学参数:Ke0.31h-1,AUC18.07μg·h·mL^-1,t_{1/22.24h,cmax6.26μg·mL^-1,tmax0.75h ;皮肤给药途径主要药动学参数:Ke0.92h^-1,AUC32.4μg·h·mL^-1,t1/2 0.75h,cmax1.17μg·mL-1,tmax1h。结论　本法分离效果好 ,分析速度快 ,方法稳定 ,结果准确。}     

茵陈蒿及其汤剂药物动力学比较研究

目的 :测定大鼠灌胃给予茵陈蒿单煎液与茵陈蒿汤后 ,茵陈蒿中药效物质成分 6 ,7 二甲氧基香豆素的血药浓度 ,研究其药物动力学行为。方法 :采用反相高效液相色谱 紫外检测法。以香豆素为内标 ,血浆样品经乙腈沉淀蛋白 ,上清液在 50°C下经氮气流吹干 ,残渣加入流动相溶解后 ,离心 ,上清液进样。流动相为甲醇 1 %乙酸溶液 四氢呋喃 (30∶63∶7) ;检测波长为 340nm。大鼠经灌胃 (4mg·kg- 1 ,按 6 ,7 二甲氧基香豆素计 )给予茵陈蒿单煎液及茵陈蒿汤后 ,所测得血药浓度数据用Topfit程序拟合并估算药物动力学参数。结果 :方法线性范围为 0 .0 2 5～ 2 .5μg·mL- 1 。经非房室模型计算 ,药物动力学参数T1 / 2 分别为 1 .30和 1 .75h ,AUC0→t分别为 1 2 1 5 .0 0和 2 52 7.85ng·h·mL- 1 ,AUC0→∝ 为 1 32 5 .90和2 61 2 .58ng·h·mL- 1 。结论 :本文建立的方法适用于 6 ,7 二甲氧基香豆素血药浓度测定及药物动力学研究。大鼠灌胃给予茵陈蒿汤后较单独给予菌陈蒿液后 ,血浆中 6 ,7二甲氧基香豆素 1 /2延长 ,AUC值显著增高     

反相高效液相色谱法测定大鼠血浆中补阳还五汤中黄芪甲苷的含量及药动学研究

 目的对大鼠血浆补阳还五汤中黄芪甲苷的含量进行了测定,并进行了药动学研究。方法采用反相高效液相色谱法测定,其条件为色谱柱:C₁₈柱(4.6mm×250mm,5μm);检测波长:203nm;流动相:乙腈-水(35:65),流速:1mL·min^-1;柱压:(15.17±0.4137)MPa;温度:45℃。药动学参数采用3P87软件处理。结果血浆样品中黄芪甲苷线性范围为2.4～12μg,r=0.9996,回收率为100.2%,RSD为1.100%。黄芪甲苷药动学参数:复方:t_1/2α为1.392h;t_1/2β为14.24h;K₁₂为0.1624h^-1;K₂₁为0.3045h^-1;K₁₀为0.07958h<>sup-1;总苷:t_1/2α为2.641h;t_1/2β为37.12h;K₁₂为0.1283h^-1;K₂₁.为0.1071h^-1;K₁₀为0.04574h^-1。结论该法快速、简便,准确。补阳还五汤复方及总苷中黄芪甲苷药动学为二室模型。中药复方成分组合对药动学参数有一定影响。     

The pharmacokinetics and oral bioavailability studies of columbianetin in rats after oral and intravenous administration

Ethnopharmacological relevance

The roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan (RAP) has been used as Traditional Chinese medicine to treat rheumatic disease in China since ancient times, but its action mechanisms was not well understood. Columbianetin is one of the main active constituents isolated from RAP, which has been shown to have various biological activities, but the absorption characteristics and oral bioavailability dose proportionality of columbianetin in vivo were not studied.

Materials and methods

Male Sprague Dawley rats (210–230 g) received either an intravenous (i.v. 5, 10 and 20 mg kg⁻¹) or oral (5, 10 and 20 mg kg⁻¹) dose of columbianetin. The levels of columbianetin in plasma were measured by a simple and sensitive reversed-phase high-performance liquid chromatography (HPLC) method. The simple liquid–liquid extraction with ethyl acetate was used for sample preparation. Osthole was selected as internal standard (IS).

Results

The chromatographic separation was accomplished on a C₁₈ column at a flow rate of 1 mL min⁻¹, where water–methanol was used as mobile phase. The calibration curve of the method was linear in the concentration range of 0.05–2000 μg mL⁻¹. The intra and inter-day accuracy for columbianetin in rat plasma samples were within 8% and the variation was less than 8.3%. This method was suitable for the determination and pharmacokinetic study of columbianetin in rat plasma after both intravenous and oral administration. The results indicated that maximum plasma concentrations(C_max) for the columbianetin (17–42 μg mL⁻¹) were achieved at 0.3–0.5 h post-oral dosing and the apparent volume of distribution (V/F) ranged from 0.38 to 0.44 L. Absolute bioavailability of columbianetin was assessed to be 81.13±45.85, 81.09±33.63 and 54.30±23.19%, respectively. Terminal elimination half-life (T_1/2) of the columbianetin after oral dosing was 60–90 min and were 2.5–3.3 fold longer than those observed for the i.v. dosing.

Conclusions

The pharmacokinetic properties of columbianetin in rat after oral administration were characterized as rapid oral absorption, quick clearance and good absolute bioavailability. The bioavailability of columbianetin ranged from 54 to 81% for 5, 10 and 20 mg kg⁻¹ oral doses. The bioavailability of columbianetin is independent of the doses studied. Columbianetin showed dose proportionality over the dose range 5–20 mg kg⁻¹. The results clearly demonstrated that columbianetin was one of the material bases of RAP. Furthermore, an HPLC method was demonstrated in this study for the research of traditional Chinese medicine.