HPLC测定六味丁香丸中的土木香内酯

目的采用RP-HPLC法测定六味丁香丸中的土木香内酯。方法采用EClipse XDB-C18色谱柱,乙腈-0.05%磷酸溶液(50∶50)为流动相,柱温40℃,流速1.0 m L·min-1,检测波长194 nm。结果土木香内酯0.293~3.51μg与峰面积的线性关系良好(r=0.9999),平均加样回收率为98.74%,RSD=0.57%(n=6)。结论所用方法准确、简便、灵敏、可靠,可用于六味丁香丸的质量控制。     

Alantolactone,a sesquiterpene lactone,inhibits breast cancer growth by antiangiogenic activity via blocking VEGFR2 signaling

Alantolactone (ALA), a sesquiterpene lactone isolated from several medicinal plants such as Inula helenium, has been identified to have attractive anticancer activity. However, its role in the inhibition of angiogenesis during tumor development remains unclear. In this study, we found ALA can inhibit the proliferation, motility, migration, and tube formation of human umbilical vein endothelial cells. ALA also restrained angiogenesis in chick embryo chorioallantoic membrane and delayed the growth of human MDA‐MB‐231 breast cancer xenograft in mice through angiogenesis inhibition. Furthermore, ALA suppressed the phosphorylation of vascular endothelial growth factor receptor 2 and its downstream protein kinase including PLCγ1, FAK, Src, and Akt in endothelial cells. Taken together, the antiangiogenic activity of ALA and its molecular mechanism are identified for the first time, indicating that ALA may be a potential drug candidate or lead compound for antiangiogenic cancer therapy.     

High body clearance and low oral bioavailability of alantolactone,isolated from Inula helenium,in rats: extensive hepatic metabolism and low stability in gastrointestinal fluids

Alantolactone (ALA) is a major bioactive sesquiterpene lactone present in the roots of Inula helenium L. (Asteraceae) which has been used widely in traditional medicine against various diseases such as asthma, cancer and tuberculosis. The pharmacologic activities of alantolactone have been well characterized, yet information on the physicochemical and pharmacokinetic properties of alantolactone and their mechanistic elucidation are still limited. Thus, this study aims to investigate the oral absorption and disposition of alantolactone and their relevant mechanisms. Log P values of alantolactone ranged from 1.52 to 1.84, and alantolactone was unstable in biological samples such as plasma, urine, bile, rat liver microsomes (RLM) and simulated gastrointestinal fluids. The metabolic rate of alantolactone was markedly higher in rat liver homogenates than in the other tissue homogenates. A saturable and concentration‐dependent metabolic rate profile of alantolactone was observed in RLM, and rat cytochrome P450 (CYP) 1 A, 2C, 2D and 3 A subfamilies were significantly involved in its hepatic metabolism. Based on the well‐stirred model, the hepatic extraction ratio (HER) was estimated to be 0.890–0.933, classifying alantolactone as a drug with high HER. Moreover, high total body clearance (111 ± 41 ml/min/kg) and low oral bioavailability (0.323%) of alantolactone were observed in rats. Taken together, the present study demonstrates that the extensive hepatic metabolism, at least partially mediated by CYP, is primarily responsible for the high total body clearance of alantolactone, and that the low oral bioavailability of alantolactone could be attributed to its low stability in gastrointestinal fluids and a hepatic first‐pass effect in rats. Copyright © 2016 John Wiley & Sons, Ltd.     

正交试验优选藏木香与干姜中挥发性成分萃取工艺

目的:优选藏木香与干姜中挥发性成分的最佳超临界CO2萃取工艺。方法以土木香内酯含量、挥发油得率为指标,以萃取压力、萃取温度、萃取时间及夹带剂用量为考察因素,采用正交试验优选最佳萃取工艺。结果:最佳萃取工艺为夹带剂量225mL,在萃取压力35Mpa下,于40℃萃取1h。结论:该工艺合理、操作可行、质量可控,可用于工业化生产。     

Bioavailability and pharmacokinetics of alantolactone from Inula helenium in rats following intravenous and oral administrations

Alantolactone, as the principal constituent of Inula Helenium L, has been shown various pharmacologic activities, such as anti-inflammatory and deworming. In the present study, we developed a high performance liquid chromatography (HPLC) method for the determination of alantolactone in rat plasma, and pharmacokinetics of alantolactone was investigated after intravenous and oral administrations to Wistar rats. Separation was achieved on C₁₈ column (4.6 mm×250 mm, 5.0 μm) using a mobile phase consisting of methanol–water (70:30, v/v) at a flow rate of 1.0 mL/min. The wavelength of the ultraviolet detector was set at 239 nm. The excellent linearity was found over a concentration range of 0.08–10 μg/mL (R² = 0.9998).The intra- and inter-day precisions were good, and the RSD was lower than 2.27%. The mean absolute recovery of alantolactone in plasma ranged from 88.09% to 95.57%. After intravenous administration, alantolactone showed rapid systemic clearance (CL (0.11±0.014) L/h/kg) and small volume of distribution (Vd (0.71±0.14) L/kg). The biological half life (t_1/2) was 56.24 min. After oral administration, alantolactone showed rapid oral absorption in rats, with a short T_max of(45.02±0.88) and (45.13±0.39) min for 14 and 28 mg/kg, respectively.The bioavailability of alantolactone in rats was 50.88%, indicating that alantolactone was orally available.     

新疆木香挥发油气相色谱—质谱分析

用ＳＥ－５４毛细管柱ＧＣ－ＭＳ联用技术分析了新疆木香挥发油的化学成分，初步鉴定了其中３１个化合物，占挥发油色谱峰总面积的７９．７８％，主要成分为土木香内酯，异土木香内酯，β－榄香烯，香橙烯，夙毛菊内酯，肉豆莞醚及二氢异土木香内酯等。     

HPLC-DAD法同时测定七珍汤散（努布炖屯汤）中4个成分

目的： 建立HPLC-DAD法同时测定七珍汤散中没食子酸、6-姜辣素、异土木香内酯、土木香内酯的含量,为七珍汤散的质量控制研究、考察其处方各味藏药材的成分和质量标准的提高提供参考。方法： 采用Luna C₁₈色谱柱（4.6 mm×250 mm,5μm）,流动相乙腈-0.2%醋酸溶液,梯度洗脱（0~10min,2%→12% A;10~20 min,12%→50% A;20~30 min,50%→85% A;30~50 min,85%→2% A）,流速1.0 mL·min^-1,柱温35℃,检测波长230、280 nm。结果： 4种成分实现完全分离;没食子酸、6-姜辣素、异土木香内酯、土木香内酯的线性范围依次分别在7.42~148.4、0.92~18.4、1.83~91.25、1.71~85.30 μg（r ≥ 0.9990）;平均加样回收率依次分别为99.06%、99.27%、99.86%、99.36%,RSD依次分别为1.33%、0.67%、0.35%、0.18%。结论： 该方法对4个成分同时测定,简便准确,重复性好,灵敏度高,阴性试验无干扰,可用于七珍汤散的质量控制。     

HPLC法同时测定康妇软膏中异土木香内酯、土木香内酯、花椒毒酚、氧化前胡素、欧前胡素和蛇床子素

摘 要 目的：建立HPLC梯度洗脱法同时测定康妇软膏中异土木香内酯、土木香内酯、花椒毒酚、氧化前胡素、欧前胡素和蛇床子素的含量。 方法: 以Agilent TC C18（250 mm×4.6 mm,5 μm）为色谱柱,甲醇 0.1%甲酸溶液为流动相,梯度洗脱,流速0.9 ml·min^-1 ,检测波长分别为220 nm和300 nm,柱温35 ℃。 结果:异土木香内酯、土木香内酯、花椒毒酚、氧化前胡素、欧前胡素和蛇床子素分别在6.16～123.20 μg·m^-1 、3.78～75.60 μg·m^-1 、1.87～37.40 μg·m^-1 、4.06～81.20 μg·m^-1 、9.27～185.40 μg·m^-1 、13.89～277.80 μg·m^-1 范围内线性关系良好,相关系数分别为0.999 4,0.999 8,0.999 6,0.999 5,0.999 9和0.999 7;平均加样回收率分别为98.04%（RSD=1.06%）,97.10%（RSD=1.53%）,96.73%（RSD=0.90%）,98.92%（RSD=1.36%）,99.12%（RSD=0.83%）和100.27%（RSD=0.58%）。结论:该方法简便、准确,可用于康妇软膏质量标准的提高。     

新疆木香中土木香内酯与异土木香内酯的分离鉴定

采用络合柱层法从新疆木香Ｉｎｕｌａ ｈｅｌｅｎｉｕｍ石油醚溶出部分分离得到两个结晶，经光谱分析鉴定为土木香内酯（Ａｌａｎｔｏｌａｃｔｏｎｅ）和异土木香内酯（Ｉｓｏａｌａｎｔｏｌａｃｔｏｎｅ）。     

气相色谱内标法同时测定土木香中土木香内酯、异土木香内酯的含量

[目的]建立内标法测定土木香药材中土木香内酯、异土木香内酯的含量.[方法]SE-30毛细管柱（30m×0.32mm×0.25μm）;FID检测器温度为260℃;进样口温度240℃程序升温：初始温度50℃,以20℃/min的速率升温至170℃,保持28min,再以25℃/min的速率升温至250℃,保持1 min;以尼泊金丁酯为内标物定量.[结果]土木香内酯与异土木香内酯分离良好,在0.6～4.2mg/ml范围内,呈良好的线性关系（r=0.9999,n=5）.[结论]本法简便、灵敏、专属性强、重复性好,可用于土木香药材的含量测定,控制其质量.