HPLC法同时测定清胃黄连丸中5种成分的含量

目的:建立同时测定清胃黄连丸中栀子苷、芍药苷、盐酸小檗碱、黄芩苷和丹皮酚含量的方法。方法:采用高效液相色谱法。色谱柱为Diamonsil C18,流动相为乙腈-0.3%磷酸(梯度洗脱),流速为1.0 ml/min,检测波长为238 nm,柱温为30℃,进样量为10μl。结果:栀子苷、芍药苷、盐酸小檗碱、黄芩苷、丹皮酚检测质量浓度线性范围分别为15.36~153.6、6.56~65.6、22.74~227.4、26.06~260.6、5.57~55.7μg/ml(r≥0.999 8);精密度、准确度、重复性试验的RSD≤1.0%;加样回收率分别为96.28%~99.38%(RSD=1.1%)、97.13%~99.48%(RSD=1.2%)、98.14%~100.25(RSD=0.7%)、97.38%~100.05%(RSD=1.0%)、96.30%~99.12%(RSD=1.2%),n均为6。结论:该方法操作简便、重复性好,适用于清胃黄连丸中栀子苷、芍药苷、盐酸小檗碱、黄芩苷和丹皮酚的含量测定。     

HPLC法测定清胃合剂中盐酸小檗碱和黄芩苷的含量

目的建立清胃合剂盐酸小檗碱和黄芩苷含量的测定方法。方法采用反相HPLC法同时测定清胃合剂中盐酸小檗碱和黄芩苷的含量。流动相为乙腈-0.1 mol·L-1磷酸二氢钾(25∶75),检测波长265 nm。结果盐酸小檗碱质量浓度为14.5~232 mg·L-1,黄芩苷质量浓度为35.5~568 mg·L-1时线性关系良好,平均回收率分别为103.15%和100.27%。结论本法测定结果准确,适用于该制剂的含量测定。     

清胃黄连片的质量标准

目的:建立清胃黄连片的质量控制标准。方法:采用薄层色谱法对清胃黄连片中黄芩、黄连、黄柏、甘草、知母、桅子、赤芍进行定性鉴别,采用高效液相色谱法对清胃黄连片中的黄芩苷进行含量测定。结果:黄芩、黄连、黄柏、甘草、知母、桅子、赤芍可在不同的薄层条件下分别检出;利用高效液相色谱法测定制剂中黄芩的有效成分黄芩苷,黄芩苷在0.20～1.60μg范围内线性关系良好(r=0.999 8),平均回收率为98.46%,RSD为2.89%(n=6)。结论:该法专属性强,重复性好,可用于清胃黄连片的质量控制。     

RP-HPLC法测定鼻炎康片中黄芩苷的含量

目的:采用高效液相色谱法测定鼻炎康片中黄芩苷的含量.方法:以C18化学键合硅胶柱分离黄芩苷,以甲醇-水-醋酸(42∶58∶1)为流动相,检测波长274nm.结果:黄芩苷峰与其他组分峰的分离度为2.5;理论塔板数以黄芩苷峰计算为19 516;方法的平均加样回收率为98.4%(n=5);5次独立测定的相对偏差RSD为1.0%;黄芩苷在0.25～2.5μg范围内浓度与吸收面积值呈良好的线性关系.结论:本法测定鼻炎康片中黄芩苷的含量,结果准确,重复性好.     

HPLC法测定止喘雾化液中黄芩苷的含量

目的:建立止喘雾化液中黄芩苷的含量测定方法。方法:采用反相高效液相色谱法固定相为十八烷基硅烷键合硅胶,以甲醇 水磷酸( 4 7∶5 3∶0 .2 )为流动相,检测波长2 77nm ,测定该制剂中黄芩苷的含量。结果:平均回收率为97.5 % ,RSD为1 .0 4 % (n =5 ) ,线性范围为0 .1～1 .6 μg ,r=0 .9996。结论:方法简便、快速、准确,适合该制剂中黄芩苷的含量测定。     

高效毛细管电泳法测定清胃黄连丸中盐酸小檗碱含量

目的建立测定清胃黄连丸中盐酸小檗碱含量的高效毛细管电泳法。方法毛细管区带电泳法,采用熔融石英毛细管柱(47cm×75μm),有效长度40 cm,缓冲溶液为pH=3.0的60 mmol/L磷酸盐溶液-甲醇(65∶35),分离电压30kV,进样时间5s,毛细管柱温25℃,紫外检测波长200 nm。结果盐酸小檗碱进样质量浓度在0.05～0.45 g/L范围内与峰面积线性关系良好(r=0.999 7),平均加样回收率为100.31%,RSD为2.53%(n=5)。结论该方法测定清胃黄连丸中盐酸小檗碱的含量灵敏、快速、结果可靠。     

反相高效液相色谱法测定复方鱼腥草片中黄芩苷含量

目的:采用反相高效液相色谱法测定复方鱼腥草片中黄芩苷的含量,以控制该制剂的质量.方法: 以C18化学键合硅胶柱分离黄芩苷,以甲醇 水 醋酸(40∶60∶1)为流动相,UV检测波长274 nm进行测定.结果: 黄芩苷峰与其他组分峰的分离度为2.1,理论塔板数以黄芩苷峰计算为15 590;方法的平均加样回收率为98.8%(n＝5);5次独立测定的相对标准差为RSD＝0.92%.黄芩苷线性范围0.25～2.50 μg,进样量与吸收面积值呈良好的线性关系.结论:该法测定复方鱼腥草片中黄芩苷的含量,结果准确,重复性好.     

唇齿清胃丸中栀子苷含量测定标准的研究

目的建立唇齿清胃丸中栀子苷含量测定的方法。方法采用HPLC法测定唇齿清胃丸中栀子苷的含量,使用Kromasil C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-水(15∶85)为流动相,流量为1.0 m L/min。结果栀子苷进样量在0.0330~0.8250μg范围内线性关系良好,且平均回收率为99.42%,RSD=1.1%(n=6)。结论该方法准确性、重复性及稳定性良好,可用于唇齿清胃丸中栀子苷的含量测定。     

高效液相色谱法测定清胃瘦身胶囊中黄芩苷含量

目的建立HPLC法测定清胃瘦身胶囊中黄芩苷的含量方法。方法色谱柱C18柱(150mm×4.6mm,5μm);检测波长为280nm,以甲醇:水:磷酸(50:50:0.2)为流动相。结果黄芩苷在0.1056～0.9504μg范围内呈良好的线性关系(R=0.9999),平均回收率97.9%,RSD1.6%。结论方法简单可行,可控制清胃瘦身胶囊的质量。     

高效液相色谱法测定强骨壮腰蜜丸中淫羊藿苷含量

目的 采用高效液相色谱法测定强骨壮腰蜜丸中淫羊藿苷的含量。方法以C18化学键合硅胶柱分离淫羊藿苷，以乙腈-水-醋酸(25∶75∶1)为流动相，UV检测波长270 nm进行测定。结果淫羊藿苷峰与其他组分峰的分离度为14.0，理论塔板数以淫羊藿苷计算9 800，平均回收率98.3%，RSD 0.6%(n＝5)，淫羊藿苷进样量与吸收面积分值呈良好的线性关系。线性范围0.25～2.5 μg。结论该方法测定强骨壮腰蜜丸中淫羊藿苷的含量，结果准确，重复性好。     

Development and in vivo evaluation of baicalin-loaded W/O nanoemulsion for lymphatic absorption

Background: Lymphatic formations that effectively eradicate the virus in the lymphatic system will be therapeutically advantagous in hepatitis B virus (HBV) infection. Lipid-based formulation is often used to deliver drug via the lymphatic system. Baicalin nanoemulsion may be a promising drug delivery system for improved treatment of HBV infection.

Objective: This study aimed to prepare, characterize, and evaluate a lipid-based nanoemulsion containing baicalin for lymphatic system absorption.

Method: The presence of a nanoemulsion region was studied by pseudoternary phase diagrams. The physicochemical properties of a baicalin-loaded nanoemulsion were investigated. The oral bioavailability of the baicalin-loaded nanoemulsion was compared to that of a baicalin suspension. A chylomicron flow blocking model was used to examine the extent of lymphatic uptake. The lymph node distribution of baicalin was measured to investigate the lymphatic transport ability of the nanoemulsion compared to the suspension.

Results: Compared to the baicalin suspension, the AUC_0-t and C_max values of the baicalin nanoemulsion were increased by 10.5-fold and 3.12-fold, respectively. Compared with the saline-treated rats that were orally administered the baicalin nanoemulsion, the AUC_0-t and C_max values of the nanoemulsion for the rats pretreated with cycloheximide were reduced from 23.076?±?1.244?mg/L h to 9.236?±?0.940?mg/L h and from 3.010?±?0.119?mg/L to 1.567?±?0.220?mg/L, respectively. In comparing baicalin in W/O nanoemulsion with suspension, the C_max value was found to be 11.5-fold higher in the lymph nodes of the rats treated with the nanoemulsion.

Conclusion: The results indicated that a baicalin-loaded W/O nanoemulsion may be a promising drug delivery system for the treatment of chronic hepatitis B.     

黄芩汤中黄芩苷的大鼠在体肠吸收特性研究

目的通过大鼠在体单向肠灌流实验研究黄芩汤中黄芩苷在体肠吸收特性。方法建立大鼠在体单向肠灌流模型,采用质量法计算黄芩苷吸收速率常数(Ka)和有效渗透系数(Peff)。结果黄芩汤中黄芩苷在十二指肠、空肠、回肠的吸收速率和程度都存在一定差异。其中黄芩苷在十二指肠的吸收速率显著高于空肠段,差异具有统计学意义(P<0.05),略高于回肠段但差异无统计学意义;黄芩苷在十二指肠的吸收程度明显高于空肠段且差异具有统计学意义(P<0.05),略高于回肠段但差异无统计学意义。结论黄芩汤中黄芩苷在十二指肠的吸收速度和吸收程度较好,表明黄芩汤中黄芩苷在体吸收具有肠道部位依赖特性。     

健康志愿者单次和多次口服美敏伪麻缓释胶囊的药动学研究

目的研究美敏伪麻缓释胶囊经单、多次给药后的药动学特征,评估其在健康志愿者体内的安全性。方法 22例受试者随机、开放试验设计,研究单、多次给药药动学特征。血浆中氯苯那敏、伪麻黄碱、右美沙芬、右啡烷采用LC-MS/MS法测定,药动学参数采用Win Nonlin软件计算。安全性特征以记录到的所有不良事件来进行评价。结果整个研究过程中没有严重不良事件报告。单次口服美敏伪麻缓释胶囊后,伪麻黄碱、氯苯那敏、右美沙芬和右啡烷均在3~5 h达峰,t1/2分别为6.30±1.17、23.3±6.91、10.4±2.19、8.62±3.04 h,Cmax分别为203±40.4、5.05±1.39、4.29±3.95、1.95±0.72 ng/m L,AUClast分别为2 050±559、137±47.5、61.3±67.5、17.2±6.58μg·h/L,AUCinf分别为2 140±570、161±63.8、17.6±6.65、62.8±69.3μg·h/L。在2次/d,连续给药4 d后基本达到稳态血药浓度,伪麻黄碱、氯苯那敏、右美沙芬和右啡烷的Cmax、Cmin、AUCtau,ss与单次给药比较均有不同程度的升高,波动系数的平均值为107%~271%。结论美敏伪麻缓释胶囊多次给药后4 d达到稳态血药浓度,暴露均较单次给药后有所增高,在健康志愿者体内安全性良好。     

镇癫开窍颗粒提取工艺研究

目的 确定镇癫开窍颗粒的最佳提取工艺。方法 应用HPLC同时测定龙胆苦苷、芍药苷、黄芩苷的含量,并以含量测定的综合评分为考察指标,采用正交试验法L₉（3⁴）优选提取工艺条件。结果 对提取工艺的影响因素从大到小依次为提取次数、提取时间、乙醇浓度、加入溶媒量,最适工艺条件为8倍量的80%乙醇,浸泡30 min,提取3次,每次60 min。结论 该工艺稳定、可行。     

和肝利胆颗粒提取工艺研究

目的：优选和肝利胆颗粒最佳提取工艺条件。方法：以加水量、提取时间和提取次数为考察因素,以黄芩苷含量和浸膏得率为评价指标,采用正交试验优选最佳工艺条件。结果：最佳工艺条件为加8倍量水,提取3次,每次2h。结论：该提取工艺稳定可行,可用于和肝利胆颗粒的提取。     

Comparative pharmacokinetics of a marker compound,baicalin in KOB extract after oral administration to normal and allergic-induced rats

Abstract

KOB extracts are a polyherbal medicine had been prescribed for the treatment of hyperhydrosis and allergic diseases such as allergic asthma and rhinitis in oriental clinics. Therefore, the pharmacokinetic studies of the KOB extract administered orally to normal rats and rhinitis-induced rats to understand the correlation of the efficacy and plasma concentration of KOB in patients of allergic rhinitis in future were performed. The study was conducted according to administration for pure baicalin in normal rats, baicalin in KOB extract in normal rats and rhinitis-induced rats. Baicalin in rat plasma was analyzed and validated by HPLC analysis. The interday precision based on the standard deviation of replicates of quality control samples ranged from 3.6% to 7.9% with accuracy ranging from 92.9% to 101.2% for baicalin. Based on validated analysis, pharmacokinetic study was carried out. Pure baicalin in normal rats and baicalin in KOB extract in normal rats showed bimodal curves due to direct absorption and glucuronidation. The T_max, C_max and AUC of pure baicalin in normal rats or baicalin in KOB extract in normal rats were 12?h, 0.68?µg/ml and 9.85?µg?h/ml, respectively, or 12?h, 0.46?µg/ml and 6.36?µg?h/ml, respectively. The analytical method showed excellent sensitivity, precision and accuracy, being successfully employed in a pharmacokinetic study of polyherbal medicine, KOB extract. Allergic-induced condition did not affect the pharmacokinetics of KOB extracts, suggesting KOB extracts did not require dosage adjustment in subjects with allergic-induced diseases.     

Inhibitory activities of baicalin against renin and angiotensin-converting enzyme

Context: Baicalin has been characterized as the active compound and quality control marker in Scutellaria baicalensis Georgi, traditionally used as a hypotensive herb.

Objectives: To investigate the inhibitory activities of baicalin against renin and angiotensin-I converting enzyme (ACE) and their molecule mechanism of interactions.

Methods: The fluorescence method using renin substrate 1(R-2932) and the spectroscopy method by Cushman were used to determine renin and ACE activities, respectively. The fluorescence quench techniques were used to characterize their interactions.

Results: The results showed that baicalin inhibited renin activity with an IC₅₀ value of 120.36 µM and inhibited ACE activity with an IC₅₀ value of 2.24?mM in vitro. The fluorescence emission of both renin and ACE were efficiently quenched by baicalin and a complete quenching was achieved at a high concentration of baicalin. Furthermore, baicalin was more effective in quenching the fluorescence of renin (K_SV?=?60?×?10³ M^?1) than ACE (K_SV?=?17.1?×?10³ M^?1). The quenching of fluorescence of renin and ACE involved static interactions, which was characterized by the formation of quencher–enzyme complex. The baicalin–renin complex formed through three-sites binding including the active site with a binding constant of 796.15?×?10¹³ M^?1, but there was only one binding site for the baicalin–ACE complex with a much smaller binding constant of 6.8?×?10⁵ M^?1.

Conclusion: The inhibition activity of baicalin against renin was a result of the formation of stable complex through multisites binding including the active site, which could explain the higher inhibitory efficiency.     

In vitro potential modulation of baicalin and baicalein on P-glycoprotein activity and expression in Caco-2 cells and rat gut sacs

Context Previous studies have shown that Scutellariae Radix, the dried root of Scutellaria baicalensis Georgi (Labiatae), has a certain inhibitory effect on P-glycoprotein (P-gp), but the effects of its main active constituents on P-gp are still ambiguous.

Objectives In vitro studies were performed to investigate the effects of its main active constituents (baicalin and its aglycone, baicalein) on the activity and expression of P-gp in intestine using Caco-2 cells and rat gut sacs.

Materials and methods In Caco-2 cell experiments, the effects of baicalin and baicalein on P-gp activity were investigated using a P-gp substrate, rhodamine 123 and non-substrate fluorescein Na, by determining their intracellular fluorescence accumulation, and their effects on P-gp expression were determined using flow cytometry. In addition, rat gut sac model was selected to investigate the effects of baicalin and baicalein on the transport of verapamil, a classical P-gp substrate. The gut sacs of male Sprague–Dawley rats were filled with 0.4?mL the test solution contained verapamil (0.2575?mg/mL) and the drugs [baicalin and baicalein, at concentrations of 1/8 IC₅₀ (59.875, 41.5?μg/mL), 1/4 IC₅₀ (119.75, 83?μg/mL) and 1/2 IC₅₀ (239.5, 166?μg/mL)], and then incubated in Tyrode’s solution for a period of time. After termination of the incubation, the incubated solution was processed for the subsequent detection.

Results According to the results of MTT assay, the IC₅₀ values of verapamil, baicalin and baicalein were 104, 479, 332?μg/mL, respectively. The obtained results from the two models were confirmed mutually. As a result, baicalin exhibited no obvious effect on intracellular accumulation of Rh-123, and almost had no effect on P-gp expression and verapamil transportation, while baicalein significantly increased intracellular accumulation of Rh-123 (p?<?0.01), down-regulated P-gp expression (p?<?0.01) and increased the transport of verapamil (p?<?0.05).

Discussion and conclusion The results indicated that baicalein may be a P-gp inhibitor, which presented obvious inhibitory effects on P-gp activity and expression level. A comparison of the structures of baicalin and baicalein indicates that the existence of glucosyl plays a decisive role in influencing the activity and expression of P-gp.     

复方鱼腥草配方合煎对黄芩苷和绿原酸成分溶出量的影响

目的:比较复方鱼腥草全方合煎与药材单煎对黄芩苷和绿原酸成分溶出量的影响.方法:采用高效液相色谱法,测定黄芩和金银花药材单煎提取物、全方合煎提取物中黄芩苷与绿原酸的含量.结果:全方合煎中的黄芩苷溶出量比黄芩单煎增加100%,相反,全方合煎中的绿原酸溶出量比金银花单煎降低25.4%.结论:复方配伍后煎煮对成分的溶出量有明显影响.     

Comparative pharmacokinetics of chlorogenic acid in beagles after oral administrations of single compound,the extracts of Lonicera japanica,and the mixture of chlorogenic acid,baicalin, and Forsythia suspense

Context: Chlorogenic acid (ChA) is the major compound in Shuang-Huang-Lian (SHL), which is mainly composed of ChA, baicalin, and Forsythia suspense Thunb Vahl.

Objective: The effects of co-existing compounds in SHL and Lonicera japanica Thunb on the absorption of ChA was investigated.

Materials and methods: According to 3?×?3 Latin-square test, ChA alone, the extracts of Lonicera japanica, or the mixture of ChA, baicalin and Forsythia suspense (ChA effective doses is 60?mg/kg) was separately given to six beagles for seven days. The oral pharmacokinetic parameters of ChA in plasma, urine and faeces were quantified by HPLC/UV and analyzed.

Results: The pharmacokinetic parameters of ChA alone, the extracts of Lonicera japanica, and the mixture of ChA, baicalin, and Forsythia suspense were as followed: C_max (2.350?±?0.483, 1.655?±?0.576, 2.332?±?0.606?μg/mL), AUC_0-∞ (6.324?±?1.853, 4.216?±?1.886, 6.074?±?1.473?μg·h/mL), t_1/2 (0.911?±?0.187, 1.204?±?0.309, 1.094?±?0.193?h), and T_max (1.861?±?0.499, 1.000?±?0.459, 1.833?±?0.279?h). Accumulative fraction excretion of ChA in urine were 0.73?±?0.55, 1.25?±?1.23, 1.05?±?0.96%, while that in faeces were 0.68?±?0.94, 0.19?±?0.40, and 1.76?±?3.57%.

Discussion and conclusion: Co-existing compounds in SHL have no effect on the absorption of ChA, while the concomitant compounds in Lonicera japanica could decrease that of ChA. ChA in Beagles might have high biological transformation.