高效液相色谱法测定风感宁散中黄芩苷的含量

[目的]建立以高效液相色谱（HPLC）法测定风感宁散中黄芩苷含量的方法。[方法]色谱柱为CLC-ODS C18（150.0 mm×6.0mm,5μm）,流动相为甲醇-水（45∶55）,流速为0.5 ml/min,检测波长为277 nm,测定样品溶液中黄芩苷含量。[结果]黄芩苷检测浓度在2.48～12.79 mg/ml范围内与峰面积积分值呈良好的线性关系（r=0.999 8）;平均回收率为98.95%,RSD=0.71%（n=9）。检测得7批样品中黄芩苷平均含量为6.02 mg/ml。[结论]HPLC法测定风感宁散中黄芩苷含量简便、快速,可用于风感宁散的质量控制。     

盐酸小檗碱、绿原酸和黄芩苷在兔体液内浓度与其体外抑菌浓度的差异研究

 【目的】测定盐酸小檗碱、绿原酸和黄芩苷在兔体液内浓度,及各成分体外最小抑菌浓度,阐明3种中药成分清热解毒功能与其抑菌作用的关系。【方法】采用反相高效液相色谱法,测定盐酸小檗碱、绿原酸和黄芩苷在兔血液和组织液内的浓度;采用试管稀释法,测定3种中药成分对大肠杆菌的体外最小抑菌浓度。【结果】盐酸小檗碱、绿原酸和黄芩苷在兔血浆内的浓度峰值分别为3.2、5.03和7.63μg?ml-1,在兔组织液内的浓度峰值分别为0.12、0.11和0.12μg?ml-1;3种中药成分的体外最小抑菌浓度分别为1.0×103、3.75×103和6.75×103μg?ml-1,其体外最小抑菌浓度明显高于它们在兔体液内的浓度峰值。【结论】盐酸小檗碱、绿原酸和黄芩苷的抑菌作用较弱,在兔体液内不能达到有效抑菌浓度,其抑菌作用并非其清热解毒功能的主要机制。     

Comparative pharmacokinetics of baicalin in plasma after oral administration of Huang-Lian-Jie-Du-Tang or pure baicalin in MCAO and sham-operated rats

A sensitive and specific HPLC method was developed to analyze baicalin in rat plasma. The author had compared the pharmacokinetics of baicalin after oral administration of HLJDT decoction or pure baicalin in MCAO and sham-operated rats. All the rats were divided into two groups, MCAO and sham-operated rats. Each group contained two subgroups: HLJDT decoction and pure baicalin subgroup. The HLJDT subgroup oral administration of HLJDT decoction extract 10.00 g/kg according to body weight (containing baicalin 400.00 mg/kg according to body weight), the pure baicalin subgroup received a gavages at a dosage of baicalin 400.00 mg/kg according to body weight too. The pharmacokinetics parameters were analyzed by kinetica. The results indicated that the pharmacokinetics of baicalin in rat plasma was non-linear and there were significant differences between different groups. No matter in MCAO or sham-operated rats, pure baicalin had shown better absorption than HLJDT decoction. Whether administration of pure baicalin or HLJDT decoction, the MCAO rats show better, quicker absorption of baicalin than sham-operated rats. It was good for baicalin to exert pharmacological effects on healed cerebrovascular diseases. The method had been applied successfully to pharmacokinetics of baicalin in rat plasma after oral administration of pure baicalin or HLJDT decoction.     

HPLC法同时测定清瘟解毒口服液中栀子苷、黄芩苷的含量

建立同时测定清瘟解毒口服液中栀子苷和黄芩苷含量的HPLC方法。采用十八烷基键合硅胶为填充剂,以0.2%磷酸溶液为流动相A,以乙腈为流动相B进行梯度洗脱,流速为1.0 mL/min,二极管阵列检测器,提取波长为238 nm,进样量为10μL。试验结果表明,栀子苷在5～250μg/mL(Y=14614X+35809,R~2=0.9999),黄芩苷在10～500μg/mL(Y=13270x+23503,R~2=1.00)均呈良好的线性关系;两者平均回收率(n=6)分别为100.14%、99.42%,RSD分别为0.87%、0.94%。该方法操作简单,准确性、重复性、精密度、耐用性良好,适用于该制剂的质量控制,为制剂标准的完善提供了科学依据。     

膜分离技术在双黄连泡腾片除杂工艺中的应用

为了考察不同孔径陶瓷膜对中药双黄连水提液除杂效果的影响,以双黄连水提液为研究对象,比较5种不同孔径的陶瓷膜(50、100、200、450、800 nm)对滤液性状、膜通量衰减、有效成分保留率及固形物去除率的影响;同时以双黄连泡腾片中黄芩苷、绿原酸和连翘苷含量为考察指标,对膜过滤除杂与传统醇沉除杂工艺进行比较。结果表明,金银花+连翘水提液经滤过孔径为450 nm的陶瓷膜过滤后,滤液澄清、透明,绿原酸、连翘苷转移率分别为90.14%和90.28%,固形物去除率为29.75%;滤过孔径为200 nm的陶瓷膜对黄芩水提调酸调碱液除杂所制备黄芩提取物中黄芩苷含量为83.03%;醇沉除杂所制备双黄连泡腾片中黄芩苷、绿原酸和连翘苷平均含量分别为42.08、1.92和3.11 mg/g,膜过滤除杂所制备双黄连泡腾片中黄芩苷、绿原酸和连翘苷平均含量分别为42.55、2.07和3.23 mg/g,且不同批次含量测定的RSD值均3%(n=3)。综上所述,膜分离技术除杂所制备双黄连泡腾片工艺稳定可靠、简便易行,且绿色、安全,显著降低生产成本,对工业化生产和推广具有指导意义。     

膜分离技术在鱼腥草芩蓝口服液制备工艺中的应用

为了考察膜分离技术对中药鱼腥草芩蓝水提液除杂效果的影响,以鱼腥草芩蓝水提液为研究对象,采用微滤、超滤和纳滤相结合的方式,对鱼腥草芩蓝水提液进行膜过滤除杂及膜浓缩工艺进行研究,并与传统水提醇沉工艺进行比较。结果显示,鱼腥草芩蓝水提液膜过滤除杂初滤、超滤和膜浓缩工艺的最佳孔径分别为:200 nm、50 kd和1.5 kd;膜过滤除杂所制备鱼腥草芩蓝口服液中绿原酸、黄芩苷平均含量分别为1.11、8.53 mg/m L,醇沉除杂工艺所制备鱼腥草芩蓝口服液中绿原酸、黄芩苷平均含量分别为0.95、5.34 mg/m L,且不同批次含量测定的RSD值均3%(n=3);膜过滤除杂工艺制备的样品进行蛋白质检测时未出现浑浊,醇沉除杂工艺制备的样品有浑浊出现;不同工艺制备的鱼腥草芩蓝口服液,稳定性均良好。综上所述,膜分离技术除杂所制备鱼腥草芩蓝口服液工艺稳定可靠、简便易行,且绿色、安全,显著降低生产成本,本文对工业化生产和推广具有指导意义。     

近红外光谱测定清热解毒口服液中绿原酸与黄芩苷的波段优选

[目的]运用近红外光谱测定清热解毒口服液中绿原酸与黄芩苷含量的波段并建立数学模型。[方法]结合移动窗口片最小二乘(MWPLS)法对波段进行了优选,分别建立测定清热解毒口服液中绿原酸与黄芩苷含量的定量分析模型,比较了不同的光谱预处理方法的效果。[结果]采用MWPLS方法对6个厂家30个样品建模选出绿原酸与黄芩苷的最优波段分别为1 254～1 306 nm和1 176～1 266nm,使用其他2个厂家10个批次样品进行检验,绿原酸和黄芩苷实测值与预测值检验均方根误差分别为1.637和0.857,相关系数分别为0.993 7和0.987 7。[结论]定量模型预测精度高,稳定性较好,可用于清热解毒口服液生产过程中的质量控制。     

微波辅助提取法提取复方中草药有效成分的研究

[目的]研究微波辅助提取法提取复方中草药的有效成分。[方法]采用乙醇回流提取法和微波辅助提取法提取复方中药中的有效成分,同时采用紫外-可见分光光度法测定了黄芩苷和葛根素的含量,并以黄芩苷得率和葛根素得率为考察指标,比较了2种不同提取方法的提取效果,同时研究了微波辅助提取法的微波作用时间对提取的影响。[结果]以不同浓度的乙醇进行回流提取时,黄芩苷和葛根素的得率随着乙醇浓度的增加先增大后减小;以微波辅助提取法进行提取时,浓度为60%乙醇提取的黄芩苷和葛根素的得率最高。当乙醇浓度为60%时,采用微波辅助提取法提取的黄芩苷和葛根素的得率均明显高于乙醇回流提取。最佳微波作用时间为15 min,此时黄芩苷得率和葛根素得率最高,分别是0.315 5和0.502 5 mg/g。[结论]采用微波辅助乙醇回流提取法可提高复方中草药中黄芩苷得率和葛根素得率。     

Baicalin attenuates pancreatic fibrosis by inhibiting TGF-β1/TAK-NF-κB signaling pathway in chronic pancreatitis mice

AIM: To explore the role of transforming growth factor-β1 (TGF-β1)/TGF-β-activated kinase (TAK)-nuclear factor-κB (NF-κB) signaling pathway in chronic pancreatitis (CP) mice and the effect of baicalin on pancreatic fibrosis in the mice. METHODS: Kunming mice (n=58) were randomly divided into 3 groups, including control group, CP group and baicalin group. The mice in CP group and baicalin group were intraperitoneally injected with 20% L-arginine. After 2 weeks of CP, the mice in baicalin group were intraperitoneally injected with baicalin (100 mg/kg, once a day). At 2 weeks, 4 weeks and 6 weeks after modeling, the mice were anesthetized and sacrificed. The morphological changes of the pancreas were observed by HE and Masson staining. The serum level of TGF-β1 was analyzed by ELISA. The expression of fibronectin (FN) and NF-κB in the pancreas was observed by immunohistochemistry staining. The protein levels of transforming growth factor-β receptor type Ⅰ (TGF-βRⅠ), phosphorylated TAK1 (p-TAK1) and NF-κB in the pancreas were determined by Western blot. The mRNA expression of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloprotease-1 (TIMP-1) was detected by real-time PCR. RESULTS: At 2 weeks, 4 weeks and 6 weeks after intraperitoneal injection of L-arginine, the pancreatic tissues were obviously injured and exhibited different degrees of fibrosis, and FN expression was significantly increased. After treatment with baicalin, the degrees of pancreatic injury and fibrosis were significantly attenuated and the expression of FN was reduced (P<0.01). Compared with control group, the protein levels of TGF-β1, TGF-βRⅠ, p-TAK1, NF-κB and TIMP-1 in the pancreas of CP group were significantly increased, and the expression of MMP-1 was decreased at each time point. In baicalin group, the protein levels of TGF-β1, TGF-βRⅠ, p-TAK1, NF-κB and TIMP-1 were significantly decreased, and the expression of MMP-1 was markedly increased at the corresponding time points compared with CP group (P<0.01). CONCLUSION: Baicalin effectively atte-nuates pancreatic fibrosis by inhibiting the activation of TGF-β1/TAK-NF-κB signaling pathway and regulating the balance of MMP-1/TIMP-1.     

不同理化因子对黄芩悬浮细胞系的影响

采用植物组织培养手段建立了稳定的黄芩细胞悬浮系,通过正交试验筛选出最佳的MS液体培养基为:NH4+/NO3-30/40 mmol/L,Ca2+3 mmol/L,pH值5.4,蔗糖3%.研究了碳源、肌醇、激素、水解酪蛋白(CH)和水解乳蛋白(LH)对悬浮系中黄苓甙含量的影响.结果表明:复合碳源、较低浓度肌醇对悬浮细胞系中黄芩甙的积累和胞外释放具有促进作用,2.0 mg/L NAA+0.5 mg/L Kt最有利于悬浮系中黄芩甙的积累;水解酪蛋白和水解乳蛋白对黄芩甙的生成没有促进作用,反而起到了一定的抑制效应.