HPLC检测苦参中苦参碱和氧化苦参碱的含量

建立HPLC法测定苦参中苦参碱和氧化苦参碱含量。色谱柱采用Aichrom NH_2(250 × 4.6mm,5 μ m),流动相为乙腈-乙醇-3%磷酸溶液(80:10:10),流速:1.0mL/min,检测波长:210nm,进样量为10μL,柱温:30℃。苦参碱在0.5～10 μ g/mL范围内有良好的线性关系(r=0.9998),氧化苦参碱在0.5～10 μ g/mL范围内有良好的线性关系(r=0.9997)。苦参碱平均加标回收率为100.42%(n=5),RSD为1.91%;氧化苦参碱平均加标回收率为100.26%(n=5),RSD为2.03%。该方法操作简单、重现性好、检测灵敏度、准确度高,且样品分离效果好。本方法适用于苦参药材中苦参碱和氧化苦参碱的含量测定。     

HPLC-MS法测定Beagle犬血浆中氧化苦参碱及其药代动力学

目的: 建立Beagle犬血浆中氧化苦参碱的LC-MS测定法,测定它的绝对生物利用度。方法: Beagle犬6只,随机分为2组,采用单剂量双周期自身交叉设计,分别给犬单剂量静脉注射或灌胃氧化苦参碱,用LC-MS法测定给药后的血浆中药物浓度。结果: 氧化苦参碱在2～5000μg·L^-1的范围内呈良好的线性关系(r=0.9990),最低检出限达0.6μg·L^-1,日内和日间误差均小于4.2 %,方法回收率大于96.7 %;氧化苦参碱的药-时数据符合二室模型,C_max为2.42±0.97μg·L^-1,T_max为1.0±0.3 h,T_1/2β为5.54±1.58h,AUC_0→∝为6.12±1.08μg·L^-1·h^-1,绝对生物利用度为(19.4±9.01)%。结论: 该法灵敏、简单,专属性强;氧化苦参碱在Beagle犬体内绝对生物利用度较低。     

H103大孔吸附树脂分离纯化苦参碱与氧化苦参碱的研究

通过对比苦参总碱在九种大孔树脂上的静态吸附,筛选出对苦参总碱吸附能力较强的H103树脂;研究了H103树脂对上述两种生物碱的吸附等温曲线及吸附动力学行为;考察了苦参总碱在H103树脂上的动态吸脱附过程,用薄层色谱法(TLC)对洗脱液跟踪分析,用高效液相色谱法(HPLC)检测苦参碱、氧化苦参碱含量,确定了动态吸脱附的过程参数。结果表明,H103树脂对苦参总碱的等温吸附可采用Freundlich方程描述;由吸附动力学曲线得H103树脂在100 min内达到平衡,具有较快吸附速度,吸附动力学规律可用二级速率方程表示;动态吸脱附过程的洗脱条件为：用30%乙醇-25%氨水（115:1,v/v）,80%乙醇梯度洗脱,氧化苦参碱先被洗脱,苦参碱随后,两种生物碱可达到很好的分离,经计算苦参碱收率为90.1%,氧化苦参碱收率为85.3%。     

Development of Inhalable Chitosan-Coated Oxymatrine Liposomes to Alleviate RSV-Infected Mice

Human respiratory syncytial virus (RSV) infection is the most important cause of acute lower respiratory tract infection in infants, neonates, and young children, even leading to hyperinflation and atelectasis. Oxymatrine (OMT), originating from natural herbs, possessed potential antivirus activity against influenza A virus, Coxsackie B3 virus, and RSV, whereas the absence of an in vivo study indicated the difficulties in overcoming the physiological obstacles. Since RSV basically replicated in lung tissue, in this study, we fabricated and characterized a chitosan (CS)-coated liposome with OMT loaded for the treatment of lethal RSV infection via inhalation. The results uncovered that OMT, as a hydrophilic drug, was liable to diffuse in the mucus layer and penetrate through the gas–blood barrier to enter systemic circulation quickly, which might restrict its inhibitory effect on RSV replication. The CS-coated liposome enhanced the distribution and retention of OMT in lung tissue without restriction from mucus, which contributed to the improved alleviative effect of OMT on lethal RSV-infected mice. Overall, this study provides a novel inhalation therapy for RSV infection, and the CS-coated liposome might be a potential inhalable nanocarrier for hydrophilic drugs to prevent pulmonary infections.     

N-Trimethyl chitosan-coated multivesicular liposomes for oxymatrine oral delivery

Background: Multivesicular liposomes (MVLs), uncoated and coated with N-trimethyl chitosan (TMC), have been studied for their potential use for drug delivery by the oral route. Method: Synthesized TMC was characterized by infrared spectroscopy, revealing the presence of trimethyl groups, and by proton nuclear magnetic resonance spectroscopy, allowing the calculation of the degree of substitution quaternization (70.2%). Oxymatrine (OM), a natural quinolizidine alkaloid used clinically for treating hepatitis B, was chosen as a model drug. The surface-modified MVLs and uncoated MVLs were characterized in vitro in terms of their shape, size, zeta potential, entrapment efficiency, coating efficiency, the stability of MVLs in polymer suspension, and the stability in simulated gastric and intestinal fluids. Results: In vivo, the area under the plasma concentration–time curve obtained from the pharmacokinetics study of TMC-coated MVLs was found to be about 3.26- and 1.96-fold higher than that of OM solution and uncoated MVLs, respectively. Conclusion: TMC-coated MVLs can be considered as a potential carrier for oral drug administration.     

氧化苦参碱的合成

以苦参碱和双氧水为原料合成氧化苦参碱。考察了原料配比、反应时间、反应温度三个主要因素对反应产率的影响,优选出最佳合成工艺。结果最佳合成工艺为:n(苦参碱)∶n(H2O2)=1∶8(摩尔比)、反应时间4 h、反应温度50~60°C。氧化苦参碱收率达95%以上。     

氧化苦参碱提取条件的优化

采用单因素实验和正交实验优化了苦参中氧化苦参碱的提取条件.结果表明,最适提取溶剂为乙醇溶液;最佳提取条件为:提取溶剂乙醇的体积分数60%、料液比1:30(g : mL)、提取温度80℃、提取时间85 min,在此条件下氧化苦参碱得率达到6.58 mg·g-1,较优化前提高了2.8倍.     

氧化苦参碱通过调控上皮间质转化抑制三阴性乳腺癌细胞侵袭转移

目的: 探讨氧化苦参碱对三阴性乳腺癌细胞MDA-MB-231生长增殖及其对迁移能力的影响和机制。方法: CCK-8活细胞计数法分析氧化苦参碱不同浓度（0、1、2、4、6、8、10、20 mg/mL）在不同时间（24、48、72 h）对三阴性乳腺癌细胞MDA-MB-231生长增殖的影响,流式细胞仪检测细胞周期和凋亡,Transwell法观察氧化苦参碱对MDA-MB-231细胞迁移和侵袭能力的影响,Q-PCR法检测EMT相关因子Snail-2和E-cadherin表达水平。结果: 与对照组比较,浓度为2、4、6、8、10、20 mg/mL的氧化苦参碱处理MDA-MB-231细胞24、48和72 h后,对细胞的生长呈时间和剂量依赖性的抑制作用。流式细胞实验提示2 mg/mL氧化苦参碱可使G0/G1期细胞数明显增加（P<0.05）,S期和G2/M期的细胞数明显减少（P<0.05）,细胞生长阻滞于G0/G1期。中、高浓度氧化苦参碱（4 mg/mL、6 mg/mL）处理12 h能明显抑制MDA-MB-231的侵袭能力,抑制Snail 2 mRNA的表达（P<0.01）,上调E-cadherin mRNA和蛋白水平的表达（P<0.01）。结论: 氧化苦参碱对三阴性乳腺癌细胞生长和迁移侵袭有抑制作用,具有潜在的抗肿瘤生长和转移作用。     

HPLC法测定氧化苦参碱的含量

用HPLC法采用C18柱分离,流动相为甲醇:3%磷酸溶液=8∶2(体积分数),检测波长220nm,流速1.0mL/min,柱温为30℃测定合成氧化苦参碱的含量。结果表明:氧化苦参碱在1～10μg范围内,进样量与峰面积呈良好的线性关系(r=0.9998),氧化苦参碱的平均回收率为99.33%(n=6),RSD为0.296%。建立的HPLC方法灵敏、准确、重现性好,操作简便,可有效测定氧化苦参碱的含量。     

同位素内标法—超高效液相色谱/串联质谱法测定六堡茶中苦参碱和氧化苦参碱残留量

目的:以广西六堡茶为例,建立超高效液相色谱—质谱/质谱(UPLC-MS/MS)分析方法测定苦参碱和氧化苦参碱残留量。方法:样品经乙腈水溶液(80%乙腈+0.2%氨水)超声提取,流动相为0.1%甲酸—水和0.1%甲酸—甲醇溶液,梯度洗脱,内标法定量。结果:苦参碱和氧化苦参碱在0.1～80.0 ng/mL质量浓度范围内呈线性,相关系数均>0.999 6,检出限均为1.0μg/kg,定量限均为3.0μg/kg,满足茶叶中苦参碱和氧化苦参碱残留量的监管判定要求。通过加标验证,回收率均为92.0%～104.7%,相对标准偏差(RSD)均<5.4%。结论:试验方法结合净化管净化(147.7 mg PSA,15.1 mg GCB,887.2 mg硫酸镁),以Kinetex~? 2.6μm Biphenyl 100?色谱柱(100 mm×3.0 mm)分离,解决了苦参碱和氧化苦参碱提取回收率低、前处理复杂的问题,使苦参碱和氧化苦参碱出峰良好,抗干扰性强。