毛连菜不同采收期中绿原酸、异绿原酸A和木犀草苷的变化规律

摘 要 目的：检测不同采收期毛连菜植物中绿原酸、木犀草苷和异绿原酸A的含量,研究不同采收期毛连菜中3种化合物的动态变化规律,为该植物的开发利用提供参考。 方法: 采用高效液相色谱法（HPLC）,以乙腈 0.1%磷酸溶液梯度洗脱,同时测定不同采收期毛连菜中绿原酸、木犀草苷和异绿原酸A的含量。 结果: 3种成分的变化趋势有一定差异,绿原酸、木犀草苷在幼苗期含量最高,成熟期降到最低,花果期再升高;异绿原酸A从幼苗期开始升高至起苔期达峰值,然后又开始下降,花果期再次升高。 结论: 不同采集期毛连菜中3种化合物富集规律不同,使用全草可在植株5~6月份起苔以前采集。     

一测多评法测定藏茴香中异绿原酸A、新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C

目的 建立一测多评法（QAMS）测定藏茴香中6种咖啡酰奎宁酸类成分含量,并验证该方法在藏茴香质量评价中应用的可行性与适用性。方法 取藏茴香粉末（过三号筛）0.5 g,精密加入70%甲醇 20 mL ,超声处理（250 W、频率 53 kHz）30 min,制备供试品溶液;采用Phenomenex GeminiR C₁₈（250 mm×4.6 mm,5 μm）色谱柱,以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,检测波长为 330 nm,体积流量 1.0 mL·min^-1,柱温 30 ℃,进行专属性、供试品提取条件、检测波长选择、色谱条件、线性关系、精密度、重复性、稳定性、加样回收率方法学考察,建立异绿原酸A、新绿原酸、绿原酸、隐绿原酸、异绿原酸 B、异绿原酸 C成分含量检测的 HPLC法;以异绿原酸 A为内参成分,分别计算新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C 5种成分的相对校正因子,分别采用3种不同色谱仪和3种色谱柱进行相对校正因子、相对保留时间耐用性考察,对藏茴香样品同时采用外标法与 QAMS 测定 6 种成分的质量分数,比较 2 种测定方法结果的差异。结果 建立的6种成分的HPLC检测方法的专属性、供试品提取条件、检测波长选择、色谱条件、线性关系、精密度、重复性、稳定性、加样回收率均符合要求;新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C的相对校正因子平均值分别是 1.362、1.257、1.335、1.470、1.134,3种不同色谱仪和 3种色谱柱对相对校正因子、相对保留时间均无明显影响;QAMS与外标法2种方法测定3批藏茴香样品中6种成分得到的结果之间无显著差异。结论 建立的QAMS简便、准确、可靠,可用于藏茴香中6种咖啡酰奎宁酸类成分——异绿原酸A、新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C的定量分析。     

壮药毛冬青配方颗粒一测多评的定量研究

目的 建立一测多评法同时测定毛冬青配方颗粒中6种酚酸类成分的含量。方法 以绿原酸为内参物,分别建立其与隐绿原酸、咖啡酸、异绿原酸B、异绿原酸A、异绿原酸C的相对校正因子,并利用该相对校正因子计算含量;同时利用外标法测定该6种酚酸的含量,比较2种测定方法的差异,验证一测多评法的准确性和可行性。结果 相对校正因子的重现性良好,8批毛冬青配方颗粒的计算值与实测值无显著性差异。结论 该方法可用于毛冬青配方颗粒中多指标成分质量评价。     

一测多评法同时测定金银花标准汤剂中7种成分的含量

目的 采用一测多评法同时测定金银花标准汤剂中7种成分（新绿原酸、绿原酸、隐绿原酸、木犀草苷、异绿原酸B、异绿原酸A及异绿原酸C）的含量。方法 采用Agilent Eclipse XDB-C₁₈色谱柱（250 mm×4.6 mm,5 μm）进行HPLC测定,以乙腈-0.1%乙酸水溶液为流动相进行梯度洗脱,流速为1.0 mL·min^-1,柱温35℃,检测波长为327 nm。以绿原酸为内标物,建立与其余6种成分的相对校正因子,实现一测多评,考察校正因子的重现性。结果 一测多评法测得的金银花标准汤剂中7种成分含量,与外标法测定结果相比较,2组数据无显著性差异。结论 所建立的一测多评法方便、准确、重现性好,可用于金银花标准汤剂的质量评价。     

牛黄清感胶囊HPLC特征指纹图谱研究及多成分含量测定

目的 建立牛黄清感胶囊HPLC指纹图谱，并测定其中7种有效成分的含量。方法 采用HPLC，色谱柱为Agilent SB-C₁₈（250 mm×4.6 mm，5 μm），以乙腈（A）-0.1%磷酸水溶液（B）为流动相进行梯度洗脱；检测波长210 nm；柱温30℃。采用"中药色谱指纹图谱相似度评价系统（2004 A版）"进行相似度分析，并对指认的7个指标成分进行定量测定研究。结果 在特征图谱研究中，共确定牛黄清感胶囊HPLC指纹图谱22个共有峰，通过与对照品比较指认其中7个共有峰分别为绿原酸、咖啡酸、木犀草苷、黄芩苷、黄芩素、汉黄芩苷和汉黄芩素，利用相似度软件对12批样品指纹图谱进行分析，各批样品相似度均>0.90。定量分析条件通过方法学验证，平均加样回收率为99.1%~104.8%，RSD为1.30%~1.88%。结论 所建立的HPLC指纹图谱和含量测定分析方法可用于牛黄清感胶囊质量控制。     

一测多评法在冬菊利咽合剂质量控制中的应用研究

目的 建立一测多评法同时测定冬菊利咽合剂中绿原酸、木犀草苷和3，5-O-二咖啡酰基奎宁酸3种成分的含量。方法 采用HPLC：Agilent TC-C₁₈（2）色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱，流速1.0 mL·min^-1，柱温30℃，检测波长334 nm；以绿原酸为内参物，建立其与木犀草苷、3，5-O-二咖啡酰基奎宁酸之间的相对校正因子（f_s∕i）；采用外标法测定冬菊利咽合剂中绿原酸的含量，通过f_s∕i计算冬菊利咽合剂中其他2种成分的含量，并将一测多评法与外标法测得的实验结果进行t检验，验证一测多评法的准确性。结果 各成分的f_s/i适用性和重复性良好，采用一测多评法和外标法的测定值无显著差异。结论 本实验所建立的一测多评法可用于冬菊利咽合剂的定量分析和质量评价。     

清脂胃舒佐餐茶HPLC法指纹图谱的研究及4个成分的含量测定

目的 建立清脂胃舒佐餐茶水溶性成分的高效液相色谱（HPLC）法指纹图谱,并测定没食子酸、绿原酸、柚皮苷、橙皮苷的含量。方法 采用HPLC法,色谱条件：Kromasil C₁₈柱（250 mm×4.6 mm, 5 μm）;流动相为乙腈-0.1%磷酸水溶液,梯度洗脱;流速为0.8 ml/min;检测波长为300 nm;柱温为35℃。结果 通过中药色谱指纹图谱相似度评价系统（2004A）,确定了清脂胃舒佐餐水溶性成分的HPLC指纹图谱共有模式,标定了26个共有峰,整体相识度为0.934~0.995;并测定了10批制剂中绿原酸、柚皮苷、橙皮苷的含量分别为0.672、0.194、1.247和0.532 mg/g,RSD分别为1.35%、1.89%、0.69%和0.79%。结论 清脂胃舒佐餐水溶性成分的HPLC指纹图谱及其中没食子酸、绿原酸、柚皮苷、橙皮苷的含量检测方法简便、稳定、科学,为本品的质量控制提供了科学依据。     

菥蓂HPLC指纹图谱的建立和2种碳苷成分的含量测定

目的：采用高效液相色谱法建立菥蓂的指纹图谱,并同时测定2种主要黄酮碳苷的含量。方法：以异荭草苷和异牡荆苷为对照品,采用HSS T3色谱柱（250 mm×4.6 mm,5 μm）;以乙腈-0.5%醋酸水溶液（15：85）为流动相,流速1 mL·min^-1,检测波长350 nm,柱温30℃;采用CHEMPATTERN化学计量学软件对结果进行处理分析。结果：建立菥蓂的指纹图谱,标定5个特征峰,30 min内菥蓂的主要色谱峰能够达到完全分离,对2个特征峰成分进行含量测定。13批菥蓂中异荭草苷和异牡荆苷的含量分别为0.015%~0.094%和0.034%~0.323%。结论：建立了菥蓂的指纹图谱,结合2种主要碳苷含量测定,为菥蓂药材的质量评价提供科学依据。     

同时测定小儿咳喘灵颗粒中4种成分的方法研究

目的：建立同时测定小儿咳喘灵颗粒中（R,S）-告依春、绿原酸、甘草苷和木犀草苷含量的方法。方法：采用高效液相色谱法,Inertsil C8-3色谱柱（4.6 mm×250 mm,5 μm）;流动相：乙腈-0.5%乙酸溶液,梯度洗脱;流速：0.6 mL·min^-1;DAD检测器,检测波长：木犀草苷327 nm、绿原酸327nm、甘草苷276 nm、（R,S）-告依春245 nm。结果：金银花中木犀草苷和绿原酸、甘草中甘草苷及板蓝根中（R,S）-告依春在各自的线性范围内与其峰面积呈良好的线性关系;平均加样回收率在99.4%~103.4%范围内,RSD均符合规定。结论：该方法准确、简便、快速,可用于小儿咳喘灵颗粒中多味药材的多个活性成分的含量测定。     

正交试验法优选“武当二号金银花”藤中4种有效成分的提取条件及其含量测定

摘 要 目的：研究“武当二号金银花”藤中主要有效成分含量测定方法,为评价其品质奠定基础。方法: 采用超声法考察提取溶剂、超声时间、超声功率及料液比4个因素对样品中芦丁和木犀草苷成分提取的影响,并使用4因素3 水平的正交试验方法优化提取条件。含量测定色谱条件：色谱柱：绿原酸为C₁₈柱(Phenomenex Luna ,250 mm×4.6 mm,5 μm);马钱苷、芦丁与木犀草苷均为苯基柱(Fortis Xi phenyl ,250 mm×4.6 mm,5 μm);流动相：绿原酸、马钱苷为乙腈-0.4%磷酸溶液（15∶85）;芦丁与木犀草苷为乙腈-0.5%冰醋酸溶液梯度洗脱;柱温：30℃;检测波长分别为327,237,354,348 nm。 结果: 提取“武当二号金银花”藤中芦丁与木犀草苷成分的最佳条件：提取溶剂均为60%乙醇;料液比均为1∶30,超声功率分别为350,250 w;超声时间分别为50,60 min。样品中有效成分绿原酸、马钱苷、芦丁和木犀草苷的平均含量分别为10.27,6.33,0.401,0.450 mg·g^-1。结论:该方法简便、准确、重现性好,为其质量控制及进一步开发利用提供参考。     

高效液相色谱法测定灯盏细辛提取物中两种有效成分的含量

目的：探讨灯盏细辛提取物中野黄芩苷、3,4-二-O-咖啡酰奎宁酸2种有效成分含量的高效液相色谱测定方法。方法：色谱柱选择Dikma Kromasil C18（250mm×4.6mm,5.0μm）,流动相选择乙腈：0.2%磷酸溶液=30∶70,检测波长为332nm,柱温为35℃。结果：野黄芩苷的线性回归方程为Y=1.554×103-0.201×102（r=0.9997）;3,4-二-O-咖啡酰奎宁酸的线性回归方程为Y=1.724×103-0.081×102（r=0.9996）;野黄芩苷在0.0341～1.3121μg范围内,3,4-二-O-咖啡酰奎宁酸在0.0592～2.3426μg范围内呈良好的线性关系。结论：采用高效液相色谱法同时测定灯盏细辛提取物中2种有效成分的含量,操作简单,精密度高,重现性好,可以作为灯盏细辛提取物中这2种有效成分的定量分析方法。     

高效液相色谱-电喷雾电离质谱法分析不同时期金银花中酸性成分及含量变化

目的考察不同时期金银花中酸性成分的含量变化。方法采用高效液相色谱-电喷雾电离质谱法对金银花中酸性成分进行分析;高效液相色谱用梯度洗脱,质谱用负离子模式。结果不同时期绿原酸、异绿原酸及其同分异构体6种成分含量有一定变化。结论本方法简单、快速、重现性好,可为道地药材的评价研究提供依据。     

氯原酸与异氯原酸薄层扫描的定量研究

本文采用薄层扫描外标两点法,测定了银翘解毒片中单味药金银花的抗菌有效成分氯原酸与异氯原酸的含量和回收率分别是:0.22%、0.56%,99.5%、99.7%。     

Rapid and sensitive HPLC-MS/MS method for quantitative determination of isochlorogenic acid B in rat plasma and its application in pharmacokinetic study

Asensitive LC-ESI-MS/MS method for determination of isochlorogenic acid B in rat plasma was developed and validated in the present study. Plasma samples were prepared by a simple protein precipitation with methanol containing resveratrol as internal standard (IS). The chromatographic separation was performed on a Zorbax SB-C₁₈ column (3.5 μm, 2.1 mm×100 mm, Agilent, USA) at a flow rate of 0.2 mL/min using methanol/water containing 0.1% formic acid (v/v) as mobile phase. The detectionwas performed on a triple quadrupole tandem mass spectrometer equipped with Electronic Spray Ion by selected reaction monitoring (SRM) of the transitions at m/z 515.3→352.9 for isochlorogenic acid B and m/z 227.1→143.1 for IS, respectively. The calibration curve of the method was linear over the range of 5–2500 ng/mL (r² = 0.9982). The intra- and inter-day precisions (R.S.D.%) were less than 12.46%, and the accuracy (R.E.%) was within ±5.80%. Isochlorogenic acid B was sufficiently stable under all relevant analytical conditions. The validated method was successfully applied to the plasma pharmacokinetic studies of isochlorogenic acid B in rats. It was found that isochlorogenic acid B had non-linear pharmacokinetic characteristics in rats within the dosage ranges from 5 to 20 mg/kg.     

Protective Properties of Laggera alata Extract and its Principle Components Against d-Galactosamine-Injured Hepatocytes

Laggera alata extract (LAE) was quantitatively analyzed, and its principle components isochlorogenic acids were isolated and authenticated. Protective properties of LAE were studied using a d-galactosamine (d-GalN)-induced injury model in neonatal rat hepatocytes and a d-GalN-induced acute liver damage model in mice. Meanwhile, the effect of isochlorogenic acids derived from LAE on d-GalN-induced hepatocyte injury were also measured in vitro. LAE at concentrations of 10-100 μg/ml significantly reduced cellular leakage of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and improved cell viability. The isochlorogenic acids (4,5-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid and 3,4-O-dicaffeoylquinic acid) at concentrations of 1-100 μg/ml also remarkably improved viability of hepatocytes. The oral treatment of LAE at doses of 50, 100 and 200 mg/kg markedly reduced the serum AST and ALT activity of mice and resulted in significant recovery of hepatocytes in liver sections.     

Two new flavonoids from Centella asiatica (Linn.)

Two new flavonoids named castilliferol 1 and castillicetin 2, as well as a known compound, isochlorogenic acid 3, were isolated from the whole plant of Centella asiatica. Isolates 1 and 2 exhibited good antioxidant activity using 2,2-diphenyl-1-picryl hydrazyl radical solution with IC₅₀ values of 23.10 and 13.30, respectively. The structures of these isolates were determined by analytical and spectral data, including 1-D and 2-D NMR spectra.     

Protective effect of isochlorogenic acid B on liver fibrosis in non‐alcoholic steatohepatitis of mice

Liver fibrosis is a common symptom of non‐alcoholic steatohepatitis (NASH) and a worldwide clinical issue. The miR‐122/HIF‐1α signalling pathway is believed to play an important role in the genesis of progressive fibrosis. Isochlorogenic acid B (ICAB), naturally isolated from Laggera alata, is verified to have antioxidative and hepatoprotective properties. The aim of this study was to investigate the effect of ICAB on liver fibrosis in NASH and its potential protective mechanisms. NASH was induced in a mouse model with a methionine‐ and choline‐deficient (MCD) diet for 4 weeks, and ICAB was orally administered every day at three doses (5, 10 and 20 mg/kg). Pathological results indicated that ICAB significantly improved the pathological lesions of liver fibrosis. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hepatic hydroxyproline (Hyp), cholesterol (CHO) and triglyceride (TG) were also significantly decreased by ICAB. In addition, ICAB inhibited hepatic stellate cells (HSCs) activation and the expressions of hepatic genes involved in liver fibrosis including LOX, TGF‐β1, MCP‐1, COL1α1 and TIMP‐1. ICAB also attenuated liver oxidative stress through Nrf2 signalling pathway. What is more, the decreased levels of miR‐122 and over‐expression of hepatic HIF‐1α could be reversed by ICAB treatment. These results simultaneously confirmed that ICAB had a significant protective effect on fibrosis in NASH by inhibiting oxidative stress via Nrf2 and suppressing multiple profibrogenic factors through miR‐122/HIF‐1α signalling pathway.     

Comparison of the choleretic properties of bile acids

The choleretic properties of cholic, chenodeoxycholic, and deoxycholic acid and their taurine and glycine conjugates were compared to their ability to form micelles. It has previously been concluded that deoxycholate has the lowest critical micellar concentration; chenodeoxycholate is slightly higher and cholic is much higher. Conjugation with glycine and taurine has little or no effect on the critical micelle concentration. Since the choleretic properties of bile salts are thought to be directly proportional to their osmotic activities, one might suspect that deoxycholic acid would be the least choleretic, chenodeoxycholic slightly more choleretic and cholic much more choleretic, with little difference between the conjugated and unconjugated forms. However, in the present study, cholic, chenodeoxycholic and taurocholic acid produced similar increases in bile flow (450–700 μl/kg) after an equimolar dose (55 μM/kg). Except for the conjugation of deoxycholic acid with taurine, conjugation of these bile acids with glycine or taurine always decreased the choleretic properties of the bile acids. Therefore, it has been concluded that there is not a good correlation between the in vitro osmotic properties of bile acids and their ability to increase bile flow.     

抗癌活性成分满五酸等三个化合物的分离和鉴定

从产于四川宜宾翼梗五味子果实中,分得抗癌活性成分满五酸(1),同时分得schizandronic acid(2)和cuparenic acid(3),结构由与标准化合物比较光谱数据和物理常数而确证。     

HPLC法测定熊去氧胆酸片有关物质的含量

[摘要]目的：建立高效液相色谱法测定熊去氧胆酸片有关物质含量的方法。方法：用十八烷基硅烷键合硅胶为填充剂，以磷酸盐缓冲液（pH3.0）-甲醇-乙腈（35:37:28）为流动相，示差折光检测器测定。结果：在选定的色谱条件下，各成分可达到很好的分离，7-酮基胆石酸、胆酸、鹅去氧胆酸、胆石酸分别在0.05~2.00，0.05~2.00，0.05~2.00，0.10~2.00mg·mL-1的范围内线性关系良好(r>0.9989)；精密度、稳定性试验的RSD均不大于0.5%；回收率分别为100.1%、99.8%、100.7%和101.9%（n=9,RSD<1.4%）。7-酮基胆石酸、胆酸、鹅去氧胆酸、胆石酸定量限分别为3.0、2.5、7.0、20μg·mL-1，检测限分别为0.8、0.7、2.0、3.5μg·mL-1。测定供试品两批，供试品一7-酮基胆石酸检出量为0.03%，胆酸和胆石酸均未检出，鹅去氧胆酸检出量为0.66%；供试品二7-酮基胆石酸、鹅去氧胆酸检出量均为0.10%，胆酸和胆石酸均未检出。结论：该方法灵敏度高，专属性强，适于测定熊去氧胆酸片中有关物质的含量。