HPLC同时测定龙胆泻肝制剂中黄芩苷、汉黄芩苷、黄芩素、汉黄芩素的含量

目的:建立同时测定龙胆泻肝制剂中黄芩苷、汉黄芩苷、黄芩素和汉黄芩素4种有效成分含量的HPLC分析方法.方法:色谱柱:Dikma Diamonsil C18柱(250 mm×4.6 mm,5μm);流动相:乙腈- 0.05％磷酸溶液,梯度洗脱;波长:278 nm.结果:黄芩苷、汉黄芩苷、黄芩素和汉黄芩素的线性范围分别为0.418～4.18μg(r=0.9999),0.2～2μg(r=0.9999),0.233～2.33 μg(r=0.9999),0.215～2.15(r=0.9998).2种制剂4种成分的平均回收率分别为98.10％ (RSD=0.59％),98.05％( RSD=1.37％),98.88％( RSD=1.44％),98.10％( RSD =0.59％)和98.40％ (RSD=0.46％),99.33％ (RSD=1.74％),99.09％( RSD=1.22％),98.82％(RSD=1.42％).结论:该方法快速、简便、准确、具有良好的重复性和回收率,可作为龙胆泻肝制剂中这4种成分的含量测定方法.     

安宫牛黄丸的临床应用及不良反应浅析

安宫牛黄丸的临床应用非常广泛,在传统的内科危急重症的基础上渗透、扩展到外科、儿科等临床多科疾病。在使用安宫牛黄丸治病时,为了适应患者的不同病情,缓解病情发展,需要与其他药物配合使用。可以根据患者的病情选择用药方式,还要根据年龄、病情等需要和体质强弱决定用药时间和剂量。安宫牛黄丸在临床应用上也会有不良反应发生。本文就安宫牛黄丸在临床上的新用途和不良反应进行阐述。     

滇黄芩再生植株中黄酮类化合物的组织化学定位及黄芩苷的含量测定

应用植物组织化学定位方法,研究了组织培养滇黄芩再生植株中黄酮类化合物的积累分布状态;并且通过HPLC对滇黄芩再生植株中黄芩苷的含量进行测定。结果表明,滇黄芩再生植株黄酮类化合物主要存在于营养器官的表皮和皮层,以及茎、叶腺毛中。同时,测定再生植株中黄芩苷的含量表明,地下部分(根)含量为0.12%;地上部分(茎、叶)含量为0.43%。     

胃灵合剂中黄芩苷的含量测定

目的:对胃灵合剂中黄芩苷的含量运用高效液相色谱(HPLC)法测定分析。方法:采用VP-ODS C18色谱硅胶分析柱为固定相,甲醇-0.3%的磷酸水溶液为流动相,流速为1 ml.min-1,紫外检测波长为280nm。结果:黄芩苷峰与其他杂质峰分离良好,0.3511μg~1.2872μg范围内,线性关系良好R=0.9999。平均回收率为99.04%,RSD=0.54%(n=9)。结论:该方法简便易行,可靠准确,专属性强,重现性好,可用于胃灵合剂的质量控制。     

滇黄芩毛状根的诱导及其黄芩苷含量测定

本文利用发根农杆菌A grobacterizum rhizogenes1.2556感染滇黄芩再生苗的茎段和叶片,建立了毛状根培养及其植株再生体系。毛状根可直接从受伤的茎、叶外植体表面产生,在无外源激素的MS固体和液体培养基上自主生长,表现出典型的发根特征。毛状根茎段的诱导率较叶片高,最高可达到14.44%;经rolB基因PCR分析和甘露碱纸电泳检测,证明Ri质粒T-DNA已整合到滇黄芩基因组中并表达;毛状根在附加6-BA2mg/L和NAA0.2mg/L的MS固体培养基上直接诱导不定芽,并在MS培养基上生根,形成再生植株。获得的毛状根系经MS液体培养基培养30d后通过HPLC都能检测到黄芩苷,其中1个转化系黄芩苷含量为2.59%,是药材黄芩的0.20倍,而从3年单位时间黄芩苷生成量计算,毛状根是药材黄芩的7.18倍。本研究建立的毛状根培养体系,将对滇黄芩转基因技术的完善和利用毛状根生产黄芩苷的生物转化提供了实验基础。     

安宫牛黄丸原方及简方对脑出血大鼠损伤保护作用的研究

探讨安宫牛黄原方及简方对脑出血大鼠的保护作用,寻找更合理的组方。方法:采用胶原酶尾状核注射法造模,观察安宫牛黄丸原方及简方对脑出血损伤大鼠脑组织匀浆中超氧化物歧化酶(SOD)、丙二醛(MDA)含量的影响。结果:安宫牛黄原方及简方能不同程度的降低谷氨酸损伤神经元MDA含量,提高SOD的活性。结论:安宫牛黄丸全方及简方可在一定程度上通过抗氧化损伤来起到脑保护作用。     

黄芩的结构与黄芩苷含量的关系

应用解剖学、组织化学定位和植物化学技术,研究了一、二年生黄芩(Scutellaria baicalensis Georgi)营养器官的结构特征与黄芩苷积累的关系.结果表明:黄芩各营养器官的结构类似于一般草本双子叶植物,黄芩苷主要分布在各器官的薄壁组织细胞内,其中以维管组织的薄壁细胞为多.器官间含量的不同表现为根>叶>茎,这与其结构特征相对应,根中维管组织所占比例最大,其中薄壁组织细胞又占主要部分,故根中黄芩苷含量最高.说明根部是黄芩苷的主要贮存部位,符合黄芩以根入药的传统.不同生长期黄芩苷含量测定结果表明,两年生营养生长前期春季根内含量最高,所以春季采挖为好.     

几种影响黄芩愈伤组织中黄芩苷含量的因素

研究碳源、生长调节物质、氮源、磷酸盐以及有机添加物对黄芩愈伤组织中黄芩苷含量和产量影响的结果表明,在(25±1)℃暗培养条件下,黄芩苷积累的最佳培养基为MS基本培养基,其中氮源浓度为60mmol·L-1(NH4+:NO3-为1:1),KH2PO4浓度为1.5mmol·L-1,附加80g·L-1蔗糖、0.3mg·L-1IAA、2mg·L-16-BA和200mg·L-1蛋白胨。在上述培养基中,黄芩愈伤组织培养40d后的黄芩苷含量达到167.4mg·g-1,产量达到4.8g·L-1,分别是对照的5.54倍和18.2倍。     

黄芩的结构与黄芩苷含量的关系

应用解剖学、组织化学定位和植物化学技术,研究了一、二年生黄芩（Scutellaria baicalensis Georgi）营养器官的结构特征与黄芩苷积累的关系。结果表明：黄芩各营养器官的结构类似于一般草本双子叶植物,黄芩苷主要分布在各器官的薄壁组织细胞内,其中以维管组织的薄壁细胞为多。器官间含量的不同表现为根〉叶〉茎,这与其结构特征相对应,根中维管组织所占比例最大,其中薄壁组织细胞又占主要部分,故根中黄芩苷含量最高。说明根部是黄芩苷的主要贮存部位,符合黄芩以根入药的传统。不同生长期黄芩苷含量测定结果表明,两年生营养生长前期春季根内含量最高,所以春季采挖为好。     

定量核磁共振波谱法测定黄芩中的黄芩苷含量研究

目的:建立核磁共振波谱法测定黄芩中黄芩苷的含量.方法:以黄芩苷作为评价黄芩质量的指标,利用超声波提取法提取黄芩苷,采用核磁共振波谱法测定黄芩中黄芩苷的含量.以吡嗪为内标,氘代二甲基亚砜(DMSO-d6)为溶剂,观测频率600 MHz,脉冲延迟时间1 s,扫描32次,采集黄芩苷的1H-NMR谱图,定量峰黄芩苷质子信号δ5...     

牛黄消炎丸"泛白"现象探析

牛黄消炎丸,为百草霜包衣丸,该药常出现"泛白"现象,严重影响了产品的质量。而引起"泛白"的物质是易溶性阳离子盐类。为解决此问题,我们对百草霜的前处理工艺进行了改进,降低了百草霜中Na、K、Ca等金属离子的含量,从而减少其"泛白"现象,最终提高了药品质量。     

HPLC法测定慢严舒柠颗粒中黄芩苷的含量

采用高效液相色谱法测定慢严舒柠颗粒中黄芩苷的含量。色谱柱为ODS(C18),流动相为V(甲醇):V(水):V(磷酸)=47.0:53.0:0.2,流速为1 mL/min,检测波长为280 nm,柱温为室温。结果表明,黄芩苷进样量在0.084-0.840μg范围内线性关系良好(r=0.999 9),平均加样回收率为98.58%(相对标准偏差RSD=1.49%,n=5)。该方法简便、快捷、准确、重现性好,可用于慢严舒柠颗粒中黄芩苷的含量测定。     

Enzymatic conversion of Baicalin into Baicalein by β-glucuronidase encapsulated in biomimetic core-shell structured hybrid capsules

In this study, Baicalein was produced through an enzymatic conversion catalyzed by β-glucuronidase (GUS) encapsulated in biomimetic alginate/protamine/silica (APSi) capsules. Experimental results indicated that the thermal and pH tolerance as well as the storage and recycling stability of GUS were significantly improved after encapsulation. Under the optimum conversion conditions (37 °C, pH 7), a high productivity of Baicalein (73%) was obtained. No loss in enzyme activity was observed after 11-day storage and 90% of the initial activity remained after 26-day storage. No appreciable loss in activity was found during 10 repeated reaction cycles. The facile encapsulation process, the high conversion efficiency and the enhanced stability set an encouraging example for converting natural compounds into high value-added functional products.     

Simultaneous Determination of Diphenyl and o-Phenylphenol in Citrus Fruits

A simple method for simultaneous determination of diphenyl and o-phenyl phenol in citrus fruits was established. Fruits were distilled with a distillable oil analyzer. The citrus fruit extract was taken from this apparatus and anthracene was added as an internal standard. Gas chromatography was carried out with a column packed with 10% FFAP and a flame ionization detector. Column temperature was increased from 150 to 210°C. The retention times of diphenyl, o-phenylphenol and anthracene were 4.2, 14.0 and 15.4 min, respectively.

This method was completed within 2.5 hr and applied to samples of grapefruits, lemons, oranges, navel oranges, ponkan, iyokan, hassaku, unshu mikan and amanatsu mikan. In the recovery tests with these fruits, diphenyl was recovered in the range from 90.1 to 96.9% and o-phenylphenol was recovered in the range from 86.5 to 99.3%.     

RP-HPLC法测定银黄冲剂中黄芩苷在大鼠体内的血药浓度及药代动力学研究

采用高效液相色谱法测定银黄冲剂中黄芩苷在大鼠血浆中的浓度,并研究其药代动力学。色谱柱为D iamonsilTMC18(250 mm×4.6 mm,5μm)柱,流动相:甲醇-磷酸二氢钠缓冲液(pH 2.6),体积比为52:48,检测波长275 nm,流速:1.0 mL/m in。结果血浆中黄芩苷在0.05~20μg/mL范围呈良好的线性关系,r2=0.9997,最低定量限为6 ng/mL。大鼠按1.6 g/kg灌胃银黄冲剂后,黄芩苷的血药浓度时间曲线符合口服吸收有滞后时间的二房室模型,主要药动学参数为t1/2:0.17 h,Vd:2.65 L,K12:5.36/h,K21:0.71/h,Ke:4.25/h,AUC:2.01μg.h/mL。HPLC测定黄芩苷的血药浓度操作便捷,灵敏度高,适用于黄芩苷的药代动力学研究。     

单细胞LDL受体与清道夫受体活性的同时测定

结合应用激光扫描共聚焦显微镜系统(LSCM)和DiI-AcLDL及BODIPY FL-LDL两种荧光配基选择性标记技术,可在单细胞水平上同时测定LDL受体和清道夫受体活性.C57BL/6J小鼠巨噬细胞用终浓度为5mg/L的 DiI-AcLDL及BODIPY FL-LDL,在37℃负载5 h左右的条件下可获得良好的标记效果.两种荧光配基选择性标记具有高度特异性,在激光共聚焦显微镜下可清晰、定量地观察细胞对LDL和AcLDL摄入,是一种灵敏度高且可定量研究LDL受体和清道夫受体功能的非同位素方法.     

Simultaneous Determination of Purine and Pyrimidine Metabolites in Hprt-Deficient Cell Lines

Genetic mutations in the purine salvage enzyme, hypoxanthine-guanine phosphoribosyltransferase (HPRT), are known to cause Lesch–Nyhan syndrome and Kelley–Seegmiller syndrome. In patients, purine metabolism is different from that of normal persons. We have previously developed a method for simultaneously determining the concentration of purine and pyrimidine nucleosides and nucleotides. This system was applied to determine the concentrations of nucleosides and nucleotides in HPRT-deficient cell lines. The amount of inosine 5′-monophosphate (IMP) was different in Lesch–Nyhan syndrome, Kelley–Seegmiller syndrome, and control cell lines. The difference in the amount of IMP confirmed the mutation of the enzyme.     

Simultaneous Single-Sample Determination of NMNAT Isozyme Activities in Mouse Tissues

A novel assay procedure has been developed to allow simultaneous activity discrimination in crude tissue extracts of the three known mammalian nicotinamide mononucleotide adenylyltransferase (NMNAT, EC 2.7.7.1) isozymes. These enzymes catalyse the same key reaction for NAD biosynthesis in different cellular compartments. The present method has been optimized for NMNAT isozymes derived from Mus musculus, a species often used as a model for NAD-biosynthesis-related physiology and disorders, such as peripheral neuropathies. Suitable assay conditions were initially assessed by exploiting the metal-ion dependence of each isozyme recombinantly expressed in bacteria, and further tested after mixing them in vitro. The variable contributions of the three individual isozymes to total NAD synthesis in the complex mixture was calculated by measuring reaction rates under three selected assay conditions, generating three linear simultaneous equations that can be solved by a substitution matrix calculation. Final assay validation was achieved in a tissue extract by comparing the activity and expression levels of individual isozymes, considering their distinctive catalytic efficiencies. Furthermore, considering the key role played by NMNAT activity in preserving axon integrity and physiological function, this assay procedure was applied to both liver and brain extracts from wild-type and Wallerian degeneration slow (Wld^S) mouse. Wld^S is a spontaneous mutation causing overexpression of NMNAT1 as a fusion protein, which protects injured axons through a gain-of-function. The results validate our method as a reliable determination of the contributions of the three isozymes to cellular NAD synthesis in different organelles and tissues, and in mutant animals such as Wld^S.