高效液相色谱质谱联用法测定射干合剂中8种中药成分的含量

目的:建立一种高效液相色谱质谱联用法同时测定黄芩苷、野黄芩苷、汉黄芩素、咖啡酸、麻黄碱、射干苷、次野鸢尾黄素、黄芩素等8种中药成分的含量及其在我院自有制剂射干合剂中的应用。方法:色谱条件:色谱柱为ZORBAX SB-C18(2.1 mm×50 mm,1.8μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,流速为0.3 ml·min-1,柱温:35℃;质谱条件:采用电喷雾离子源(ESI),多反应离子监测(MRM),结合正负离子扫描切换,其中咖啡酸、野黄芩苷、射干苷采用负离子模式检测,黄芩苷、汉黄芩素、次野鸢尾黄素、黄芩素、麻黄碱采用正离子模式检测。结果:黄芩苷、野黄芩苷、汉黄芩素、咖啡酸、麻黄碱、射干苷、次野鸢尾黄素、黄芩素的定量限分别为1.44×10-4,4.20×10-3,2.95×10-4,7.80,4.90×10-3,4.6×10-2,3.18×10-4,4.85 ng·ml-1,检测限分别为4.32×10-5,1.3×10-3,8.84×10-5,0.77,2.90×10-4,3.33×10-4,9.5×10-5,1.46 ng·ml-1;在相应的线性范围内R2>0.992 3;日内和日间精密度(RSD)均小于5%,平均回收率均在80%~120%。结论:本方法在20 min内实现这8种化合物的分离和测定,简单、快速、灵敏、准确,可用于射干合剂多成分含量测定。     

射干主根与须根中6种异黄酮类成分的含量比较

目的用HPLC法测定射干主根与须根中射干苷、野鸢尾苷、鸢尾黄素、野鸢尾黄素、次野鸢尾黄素及白射干索6种异黄酮类成分的含量。方法色谱柱：KromasilC18（250mm×4．6mm,5μm）;流动相：0．2％磷酸水溶液（A）和乙腈（B）,梯度洗脱,检测波长265nm,柱温：30℃,流速1．0mL·min-1。结果射干主根中射干苷和鸢尾黄素的含量远远高于须根,主根和须根中野鸢尾苷、野鸢尾黄素和白射干素的含量相差不大（主根略高于须根）,而须根中次野鸢尾黄素的含量稍高于主根。结论该方法能准确简便地分析射干中6种成分的含量,可为射干药材的合理用药提供一定的科学依据。     

HPLC法测定射干合剂中多指标成分的含量

目的建立能同时检测射干合剂样品中5种黄酮类化合物及麻黄碱含量的HPLC法。方法样品用超声提取法,用Inertsi PODS-3色谱柱分离样品,流动相为乙腈与1‰磷酸（pH=2．25）,梯度洗脱,流量为1mL·min^-1,柱温为室温,用紫外检测仪215am测定;外标法定量。结果对于待测组分的r均〉0．998;黄芩苷、黄芩素、汉黄芩素、射干苷、次野鸢尾黄素、麻黄碱的线性范围均为0．05—50mg·L^-1.方法加样回收率为94．5％-103．2％;相对标准偏差（RSD）低于4．7％（n=3）。结论本法可用于射干合剂中多指标成分的含量测定,方法简便、准确、灵敏度高,也可用于制剂及体内指标成分的研究。     

黄芩射干汤中黄酮类成分指纹图谱的相关性

目的建立黄芩射干汤中黄酮类成分HPLC指纹图谱,研究黄芩、射干药材与复方指纹图谱的相关性。方法采用HPLC法建立黄芩射干汤的指纹图谱,以色谱峰保留时间相对偏差和紫外光谱相似性为考察指标,探讨全方指纹图谱与处方中黄芩、射干药材相关性,并对共有指纹峰进行化学成分确认。结果共得到23个色谱峰,确定了特征峰的药材归属,指认了其中11个化学成分。其中7个色谱峰来源于射干药材,分别是芒果苷、射干苷、野鸢尾苷、鸢尾黄素、野鸢尾黄素、次野鸢尾黄素和白射干素;4个峰来源于黄芩药材,分别是黄芩苷、千层纸素A-7-O-葡萄糖醛酸苷、黄芩素和汉黄芩素。结论建立的指纹图谱可为黄芩射干汤的质量控制提供依据。     

一测多评法测定川射干中8个异黄酮类成分的含量

目的:建立同时测定川射干中8个异黄酮成分(射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾黄素、鸢尾甲黄素B、鸢尾甲黄素A、野鸢尾黄素)的一测多评含量测定方法(QAMS法)。方法:采用高效液相色谱法,使用Agilent ZORBAX SB-C₁₈色谱柱(250 mm×4.6 mm, 5μm),以乙腈为流动相A,以水(含0.55%甲基-β-环糊精和0.1%磷酸)为流动相B,以0.1%磷酸为流动相C,进行梯度洗脱,流速为1.0 mL·min^-1,检测波长为266 nm,柱温为35℃。以鸢尾黄素为内参照物,采用多点校正法,分别建立射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾甲黄素B、鸢尾甲黄素A、野鸢尾黄素7个待测组分与鸢尾黄素的相对校正因子。采用校正因子计算8批川射干样品中7个待测物的含量,与外标法进行比较,验证QAMS法的准确性和可行性。结果:8个待测组分的线性范围分别是25.00～800.00、2.50～80.00、3.06～100.00、1.53～50.00、6.12～200.00、1.53～50.00、1.53～50.00、1.53～50...     

UPLC法测定射干药材中10个异黄酮类成分的含量

目的:建立同时测定射干药材中10个异黄酮类成分含量的测定方法,并用于评价不同产地射干药材中各成分的含量差异。方法:采用超高效液相色谱(UPLC)法。色谱柱为Waters ACQUITY UPLC BEH C18,流动相水相组成为含0.5%甲基-β-环糊精、0.1%磷酸的水溶液,有机相为乙腈,梯度洗脱,流速为0.2 m L/min,柱温为35℃,检测波长为265 nm,进样量为2μL,分析时间为20 min。对来自于8个省份的26个样品中的10个异黄酮类成分射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾黄素、鸢尾甲黄素B、鸢尾甲黄素A、野鸢尾黄素、次野鸢尾黄素、白射干素进行含量测定。结果:射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾黄素、鸢尾甲黄素B、鸢尾甲黄素A、野鸢尾黄素、次野鸢尾黄素、白射干素检测质量浓度线性范围分别为8.569 5～342.78、0.643～25.72、1.119 8～44.79、2.187 8～87.51、0.770 3～30.81、0.421 3～16.85、0.288 5～11.54、1.795 3～71.81、0.560 8～22.43、0.086～3.44μg/mL(r均≥0.999 6),定量限分别为0.015、0.102、0.096、0.013、0.036、0.088、0.102、0.019、0.067、0.092μg/m L;精密度、稳定性(24 h)、重复性试验的RSD均<2.00%(n=6);加样回收率为95.30%～103.30%(RSD均≤2.33%,n=6)。在26个射干样品中,以射干苷含量最高(3.66%～57.79%),白射干素含量最低(0.09%～0.59%),次野鸢尾黄素含量为0.29～2.80 mg/g,且不同产地中各异黄酮类成分含量差异较大。结论:建立的含量测定方法灵敏、分析时间较短、重复性较好,可以用于同时测定射干药材中10个异黄酮类成分的含量及评价各成分含量差异。     

UHPLC—MS／MS法在医院制剂多成分含量测定中的应用

射干合剂和甘地胶囊是新华医院的两个院内中药复方制剂,本文建立了UHPLC—MS／MS方法同时测定这两个制剂中的麻黄碱、咖啡酸、阿魏酸、芦丁、野黄芩苷、射千苷、黄芩苷、黄芩素、黄芪甲苷、次野鸢尾黄紊、汉黄芩素等11种中药成分的含量。色谱柱采用ZORBAXSB-C18（2．1mm×50mm,1．8μm）,流动相为0．1％甲酸水溶液-乙腈,梯度沈脱,流速为0．3mL／min,柱温35℃;质谱采用电喷雾离子源（ESI）,多反应离子监测（MRM）,并结合正负离子扫描切换,其中咖啡酸、阿魏酸、野黄芩苷、射干苷采用负离子模式检测,麻黄碱、芦丁、黄芩苷、黄芩素、黄芪甲苷、次野鸢尾黄素、汉黄芩素采用正离子模式检测。结果显示麻黄碱、咖啡酸、阿魏酸、芦丁、野黄芩苷、射干苷、黄芩苷、黄芩素、黄芪甲苷、次野鸢尾黄素、汉黄芩素的定量限分别为4．90×10^-3ng／mL、7．80ng／mL、6．8ng／mL、5.3×10^-2ng／mL、4．20×10^-3ng／mL、4．6×10^-2ng／mL、1．44×10^-4ng／mL、4．85ng／mL、0．23ng／mL、3．18×10^-4ng／mL、2．95×10^-4ng／mL,检测限分别为2．90×10^-4ng／mL、0．77ng／mL、2．0ng／mL、0．016ng／mL、1．3×10^-3ng／mL、3．33×10^-4ng／mL、4．32×10^-5ng／mL、1．46ng／mL、0．07ng／mL、9．5×10^-5ng／mL、8．84×10^-5ng／mL。在相应的线性范围内R^2〉0．99;日内和日问精密度（RSD）均小于5％,平均同收率均在80％-120％。本方法在20min内实现这11种目标化合物的分离和测定,简单、快速、灵敏、准确,可用于射干合剂和甘地胶囊的指标成分含量测定,为这两个制剂的质量控制提供依据。     

高效液相色谱法测定射干利咽口服液中射干苷、次野鸢尾黄素的含量

目的建立高效液相色谱同时测定射干利咽口服液中射干苷、次野鸢尾黄素含量的方法。方法采用Dia-monsil C18（4.6 mm×250 mm,5μm）色谱柱,流动相为乙腈（A）-0.1%磷酸（B）（pH=2.25）,进行梯度洗脱,0 min：20%（A）;20 min：60%（A）;40 min：20%（A）;流速：1.0 mL·min^-1;进样量：20μL;紫外检测波长：265 nm;柱温：室温。结果射干苷和次野鸢尾黄素保留时间分别为20.5和35.5 min,与各自相邻峰的分离度均〉1.5。以峰面积对进样浓度（ng·mL^-1）线性回归,射干苷回归方程：Y=7 485.5X＋82.95,r=0.999 7,线性范围：150-3 000 ng·mL^-1;次野鸢尾黄素回归方程：Y=2 031X-78.14,r=0.999 9,线性范围：50-1 000 ng·mL^-1。射干苷和次野鸢尾黄素的回收率分别为97.2%和98.7%、RSD分别为2.1%和2.8%。结论本方法操作简便,测定结果准确可靠,可用于射干利咽口服液中射干苷、次野鸢尾黄素的含量测定。     

射干异黄酮成分对5-脂氧合酶的作用研究

目的:考查射干中鸢尾苷、野鸢尾苷、鸢尾黄素、野鸢尾黄素、次野鸢尾黄素及白射干素单体成分对脂氧合酶5-LOX的影响。方法:采用脂氧合酶5-LOX抑制剂体外筛选模型进行,计算各成分对5-LOX酶的抑制率。结果:各单体成分对5-LOX均有一定的抑制作用,其中野鸢尾黄素抑制率近100%,鸢尾黄素91.7%,野鸢尾苷50%。结论:射干异黄酮单体可能通过抑制5-LOX途径发挥抗炎作用。     

8种黄酮类成分的LC-MS/MS分析

目的：为了探讨液相色谱-质谱联用（LC-MS）技术在天然产物成分分析中的应用,以黄芩苷、野黄芩苷、汉黄芩素、黄芩素、射干苷、次野鸢尾黄素、芦丁、金丝桃苷等8个黄酮类化合物为研究对象,采用超高压液相色谱-三重四级杆串联质谱分析该8个黄酮类化合物的色谱、质谱特性.方法：色谱柱为ZORBAX SB-C18（2.1 mm×50 mm,1.8 μm）,流动相为0.1％甲酸水溶液-乙腈,梯度洗脱,流速为0.3 ml·min-1,柱温为35℃,质谱离子源采用电喷雾离子源（ESI）.结果：8个黄酮化合物在色谱柱上的色谱保留行为与取代羟基及苷化的数目具有相关性,在质谱中野黄芩苷、射干苷、金丝桃苷在负离子模式下具有较好的响应,而黄芩苷、汉黄芩素、黄芩素、次野鸢尾黄素、芦丁在正离子模式下具有较好的响应.苷元的裂解以RDA裂解为主.结论：该8个黄酮化合物的色谱和质谱行为对于未知结构的黄酮化合物的结构解析具有一定的指导意义.     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Polymorphisms in genes involved in neurotransmission in relation to smoking

Smoking behavior is influenced by both genetic and environmental factors. The genetic contribution to smoking behavior is at least as great as its contribution to alcoholism. Much progress has been achieved in genomic research related to cigarette-smoking within recent years. Linkage studies indicate that there are several loci linked to smoking, and candidate genes that are related to neurotransmission have been examined. Possible associated genes include cytochrome P450 subfamily polypeptide 6 (CYP2A6), dopamine D₁, D₂, and D₄ receptors, dopamine transporter, and serotonin transporter genes. There are other important candidate genes but studies evaluating the link with smoking have not been reported. These include genes encoding the dopamine D₃ and D₅ receptors, serotonin receptors, tyrosine hydroxylase, trytophan 2,3-dioxygenase, opioid receptors, and cannabinoid receptors. Since smoking-related factors are extremely complex, studies of diverse populations and of many aspects of smoking behavior including initiation, maintenance, cessation, relapse, and influence of environmental factors are needed to identify smoking-associated genes. We now review genetic polymorphisms reported to be involved in neurotransmission in relation to smoking.     

Tramadol concentrations in blood and in cerebrospinal fluid in a neonate

Based on blood and cerebrospinal fluid samples collected in a full-term neonate, the penetration of tramadol in the central nervous system is described. Following intravenous administration of tramadol, a lag time of about 4 h was observed until full blood–brain equilibration was achieved. This pharmacokinetic observation is in line with a recent pharmacodynamic evaluation of the central opioid effects of tramadol in adults.     

Disease control in asthmatic children seen in private practice in Switzerland

ABSTRACT

Background: Asthma is the most common chronic childhood disease in Switzerland with a prevalence of 10%. Asthma has a high economic burden accounting for high medical costs. Assessment of disease control is likely to be of help in the implementation of strategies to improve asthma. Therefore, we aimed to evaluate asthma control and therapy regimens among children in private practice.

Methods: We assessed asthma control as well as therapy regimens in 575 asthmatic children in an experience programme in Switzerland by using an abbreviated questionnaire based on the asthma control questionnaire and the child health questionnaire on Visit 1 and Visit 2.

Results: Good asthma control at Visit 1 was only present in 25.7% of asthmatic children. Occasional asthma symptoms, limitation of physical activity, nocturnal awakening and anxiety of the parent was present in 80.5%, 41.2%, 46.8% and 57% of the children, respectively. After adjustment of therapy regimens at Visit 1, mainly by adding a leukotriene receptor antagonist, asthma control was reported to be much better in 53.4% of the children at Visit 2.

Conclusions: As asthma control is inadequately achieved within a major portion of asthmatic children, it is imperative to find measures to improve asthma control and hence, to reduce the burden of disease.