HPLC法同时测定产灵丸中4种有效成分

目的建立同时测定产灵丸(防风、苍术、白术等19味药物)中升麻素苷、5-O-甲基维斯阿米醇苷、苍术素醇和苍术素4种成分的高效液相色谱法。方法色谱柱为依利特C18柱(250 mm×4.6 mm,5μm);以乙腈-水为流动相进行梯度洗脱;流速为1.3 mL/min;检测波长:254 nm(升麻素苷、5-O-甲基维斯阿米醇苷)、337 nm(苍术素醇、苍术素);柱温为30℃。结果升麻素苷、5-O-甲基维斯阿米醇苷、苍术素醇和苍术素质量浓度分别在5.22~104.40μg/mL(r=0.999 3)、4.95~99.00μg/mL(r=0.999 7)、4.89~97.80μg/mL(r=0.9995)、6.34~126.80μg/mL(r=0.999 2)时与其峰面积线性关系良好;平均加样回收率分别为98.79%、97.21%、100.07%、96.90%,RSD(n=6)分别为1.19%、1.13%、0.89%、0.85%。结论该定量测定方法简便、结果准确可靠并且重现性好,可作为产灵丸中升麻素苷、5-O-甲基维斯阿米醇苷、苍术素醇和苍术素的质量控制。     

HPLC梯度洗脱法同时测定复方荆芥熏洗剂中的升麻素苷、5-O-甲基维斯阿米醇苷、齐墩果酸和熊果酸含量

目的采用高效液相梯度洗脱法建立复方荆芥熏洗剂中升麻素苷、5-O-甲基维斯阿米醇苷、齐墩果酸和熊果酸含量测定方法。方法 Hypersil C18色谱柱(4.6 mm×250 mm,5μm);流速:1.1 mL·min-1;流动相A为乙腈-甲醇(2:1),流动相B为1%冰醋酸溶液,梯度洗脱;检测波长分别为254 nm(升麻素苷和5-O-甲基维斯阿米醇苷)和205 nm(齐墩果酸和熊果酸)。结果升麻素苷、5-O-甲基维斯阿米醇苷、齐墩果酸和熊果酸分别在0.028 4⁰.568 0μg(r=0.999 7)、0.022 60.568 0μg(r=0.999 7)、0.022 6⁰.452 0μg(r=0.999 8)、0.010 80.452 0μg(r=0.999 8)、0.010 8⁰.216 0μg(r=0.999 3)、0.035 80.216 0μg(r=0.999 3)、0.035 8⁰.716 0μg(r=0.999 5)进样量与峰面积呈良好的线性关系,平均加样回收率分别为98.2%、98.0%、96.9%、97.7%,RSD(n=6)分别为0.85%、0.54%、0.93%、1.4%。结论方法简便、准确、灵敏、重复性好,可作为复方荆芥熏洗剂中升麻素苷、5-O-甲基维斯阿米醇苷、齐墩果酸和熊果酸的含量控制方法。     

关节熏洗汤剂中升麻素苷和5-O-甲基维斯阿米醇苷的含量测定

目的：测定关节熏洗汤剂中升麻素苷和5-O-甲基维斯阿米醇苷含量。方法：采用高效液相色谱法。色谱柱：Agilent Zorbax SB-C_（18）柱（250 mm×4.6 mm,5μm）;流动相：甲醇-乙腈-水（12：10：78）;检测波长：254 nm;流速：1 ml·min~（-1）;柱温：30℃。结果：升麻素苷和5-O-甲基维斯阿米醇苷的线性范围分别为0.035~0.350μg和0.037~0.370μg（r分别为0.999 9和0.999 8）,平均回收率分别为101.49%（RSD=2.5%）、98.77%（RSD=2.2%）。结论：升麻素苷和5-O-甲基维斯阿米醇苷的含量测定为关节熏洗汤剂的疗效提供了初步依据。     

RP-HPLC梯度洗脱测定至圣保元丸中升麻素苷和5-O-甲基维斯阿米醇苷的含量

目的 测定至圣保元丸中升麻素苷和5-O-甲基维斯阿来醉苷含量.方法 样品用甲醇提取,采用反相高效液相色谱法,色谱条件:Luna C18柱(250 nm×4.60 mm,5 μm)为分析柱,甲醇-乙腈-0.04%醋酸铵为流动相进行梯度洗脱;检测波长254 nm,流速1.0ml·min-1;对4批样品进行了主成分升麻素苷和5-O-甲基维斯阿米醇苷的总量测定.结果 每克至圣保元丸中升麻素苷、5-O-甲基维斯阿米醇苷的含量分别为0.159 mg,0.11 mg.回收率分别为104.16%,95.60%.结论 本方法测定至圣保元丸中的升麻素苷、5-O-甲基维斯阿米醇苷的含量准确、快速.     

高效液相色谱波长切换法同时测定表实感冒颗粒中6种成分的含量

目的建立HPLC波长切换法同时测定表实感冒颗粒中桂皮醛、3'-羟基葛根素、葛根素、升麻素苷、升麻素和5-O-甲基维斯阿米醇苷的含量。方法采用Agilent TC C18色谱柱(4.6 mm×250 mm,5μm);流动相A:乙腈-甲醇(1∶1),流动相B:0.1%磷酸溶液,梯度洗脱,流速1.0 mL·min~(-1);柱温30℃;检测波长分别为290 nm(桂皮醛)和254 nm(3'-羟基葛根素、葛根素、升麻素苷、升麻素和5-O-甲基维斯阿米醇苷)。结果桂皮醛、3'-羟基葛根素、葛根素、升麻素苷、升麻素、5-O-甲基维斯阿米醇苷分别在1.99～49.75、2.37～59.25、4.26～106.50、3.62～90.50、1.06～26.50、1.79～44.75μg·mL~(-1)与峰面积线性关系良好(r≥0.9992),6种成分的回收率均大于96.0%,RSD均小于2.0%。结论该方法操作便捷、重复性好,可用于表实感冒颗粒的质量控制。     

HPLC法同时测定舒筋通络酒中两种有效成分的含量

目的:建立高效液相色谱法同时测定舒筋通络酒中升麻苷和5-O-甲基维斯阿米醇苷含量的方法。方法:色谱柱为Lichrospher C18(150 mm×4.6 mm,10μm);流动相为乙腈-0.05%磷酸(12∶88);检测波长为254 nm;柱温为40℃;流速为1.0ml.min-1。结果:升麻苷在0.098～0.491μg范围内线性关系良好(r=0.999 9);5-O-甲基维斯阿米醇苷在0.135～0.675μg范围内线性关系良好(r=0.999 8),平均回收率分别为98.41%(RSD=1.84%),99.28%(RSD=1.56%)。结论:本法简单、方便、准确,可测定该制剂升麻苷、5-O-甲基维斯阿米醇苷的含量。     

左乙拉西坦pH敏感鼻用凝胶剂在大鼠脑组织内生物利用度的研究

目的研究左乙拉西坦pH敏感鼻用凝胶剂在大鼠脑组织内的药动学及其生物利用度。方法采用HPLC-UV法测定大鼠脑组织中左乙拉西坦浓度;以1.0%左乙拉西坦溶液灌胃给药为参比,测定该药物pH敏感鼻用凝胶剂经鼻腔给药后在脑组织中的药物浓度并计算t_(max)、C_(max)、AUC_(0～t)、AUC_(0～∞)及相对生物利用度(F_r)等主要药动学参数。结果给药剂量为1.0mg·kg~(-1)的左乙拉西坦的pH敏感鼻用凝胶与10.0 mg·kg~(-1)的左乙拉西坦溶液分别经鼻腔和灌胃给药后,测得药物在脑组织中药动学参数t_(max)分别为(0.625±0.25)和(1.25±0.50)h,C max分别为(10.60±2.52)和(19.75±2.88)μg·g~(-1),AUC_(0～t)分别为(22.84±2.73)和(50.71±6.08)μg·h·g~(-1),AUC_(0～∞)分别为(24.57±2.81)和(51.54±6.09)μg·h·g~(-1)。求得左乙拉西坦的pH敏感鼻用凝胶在脑组织中的Fr为476.83%。结论左乙拉西坦的pH敏感鼻用凝胶经鼻腔给药后,药物入脑速度快,生物利用度高,有较强脑靶向性。     

辛防通窍片中升麻素苷和5-O-甲基维斯阿米醇苷的含量测定

目的建立同时测定辛防通窍片中升麻素苷和5-O-甲基维斯阿米醇苷含量的高效液相色谱法。方法采用资生堂MG C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-水(16∶84)为流动相,检测波长为254 nm,流速为1.0 mL/min,进样量为10μL,柱温为25℃。结果升麻素苷、5-O-甲基维斯阿米醇苷进样量分别在0.151 2～6.048 0μg(r=0.999 99,n=8)和0.060 0～2.400 0μg(r=1.000 00,n=8)范围内与相应峰面积有良好线性关系,平均回收率分别为101.79%(RSD=1.44%)和101.32%(RSD=1.40%)。结论该法操作简便、结果准确、专属性强,可用于辛防通窍片的质量控制。     

HPLC法测定防风通圣丸中升麻素苷和5-O-甲基维斯阿米醇苷的含量

目的建立高效液相色谱法测定防风通圣丸中升麻素苷和5-O-甲基维斯阿米醇苷含量的方法。方法色谱柱为Inertsil ODS-3(C18)(4.6 mm×250 mm,5μm);流动相为甲醇-醋酸-水(35∶4∶61);检测波长为254 nm;流速为1.0 mL·min-1;柱温为室温。结果升麻素苷进样量在0.147 2～0.736 0μg之间,5-O-甲基维斯阿米醇苷进样量在0.134～0.670μg之间与峰面积呈良好的线性关系(相关系数r分别为0.999 7、0.999 8);二者的平均回收率分别为99.78%和99.86%。结论该方法简便、准确,专属性好,可用于防风通圣丸的质量控制。     

高效液相色谱梯度洗脱法同时测定伤科跌打丸中4种有效成分的含量

目的采用高效液相梯度洗脱法测定伤科跌打丸中异鼠李素-3-O-新橙皮苷、香蒲新苷、升麻素苷和5-O-甲基维斯阿米醇苷的含量。方法 Hypersil C18色谱柱(4.6 mm×200 mm,5μm);流速:0.9 mL·min-1;流动相A为乙腈,流动相B为0.05%磷酸溶液梯度洗脱;检测波长254 nm。结果异鼠李素-3-O-新橙皮苷、香蒲新苷、升麻素苷和5-O-甲基维斯阿米醇苷分别在0.031 2⁰.624 0μg(r=0.999 7)、0.023 80.624 0μg(r=0.999 7)、0.023 8⁰.476 0μg(r=0.999 3)、0.015 80.476 0μg(r=0.999 3)、0.015 8⁰.316 0μg(r=0.999 6)、0.010 20.316 0μg(r=0.999 6)、0.010 2⁰.204 0μg(r=0.999 4)进样量与峰面积线性关系良好,平均加样回收率分别为98.4%、97.7%、98.5%、97.7%,RSD(n=6)分别为1.1%、0.82%、0.89%、0.79%。结论该含量测定方法结果准确、灵敏、重复性好。     

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.     

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 x kg(-1)) or i.p. (50 mg x kg(-1)) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) 1 x h(-1) x kg(-1) in the male rat and 10.6 (95% CI: 7.5, 15.0) 1 x h(-1) x kg(-1) in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was approximately 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.     

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.     

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。     

Changes in angiopoietin expression in glomeruli involved in glomerulosclerosis in rats with daunorubicin-induced nephrosis

AIM: To study the potential pathological role of endogenous angiopoietins in daunorubicin-induced progressive glomerulosclerosis in rats. METHODS: Seventy male Wistar rats were allocated randomly into a daunorubicin group (DRB; n=40) or a control group (n=30). The rats in the DRB group were injected with DRB (15 mg/kg), in their tails. Subsequently, at intervals of 1, 2, 4, 6, 8, and 12 weeks, 5 male Wistar rats in each group were chosen randomly for 24 h urinary protein quantitative measurements (24 h UPQM), and determination of plasma tumor necrosis factor alpha (TNF-alpha), angiopoietin-1 (Ang1), and angiopoietin-2 (Ang2) levels. Kidney sections were examined by electron microscopy, Periodic Acid Schiff (PAS) staining, immunohistochemical staining and in situ hybridization histochemistry. RESULTS: As glomerulosclerosis progressed in the DRB group, expression of Ang1 mRNA and protein in glomeruli decreased and expression of TNF-alpha protein, Ang2 mRNA and protein in glomeruli increased. Expression of Ang1 mRNA and protein in glomeruli were negatively correlated with 24 h UPQM, Fn protein expression, and mean area of extracellular matrix (MAECM). In comparison, expression of Ang2 mRNA and protein in glomeruli were positively correlated with 24 h UPQM, Fn protein expression and MAECM; furthermore, there was a positive correlation between plasma Ang2 and 24 h UPQM. Plasma TNF-alpha and expression of TNF-alpha in glomeruli were positively correlated with expression of Ang2 mRNA and protein in glomeruli. There was a negative correlation between Ang1 protein expression and Ang2 protein expression in glomeruli. CONCLUSION: During DRB-induced glomerulosclerosis, podocyte injury led to a shift in the balance of Ang1 and Ang2 in glomeruli. Increased TNF-alpha in plasma and glomeruli may upregulate Ang2 expression in glomeruli. Elevated Ang2 in both plasma and glomeruli may mediate protein permeability through the glomerular filtration barrier. Moreover, local expression of Ang2 may facilitate the progress of glomerulosclerosis by upregulating a component expression of extracellular matrix.     

Deoxynivalenol in cereals in Russia

A survey of the occurrence of deoxynivalenol (DON) and zearalenone (ZEN) in wheat, rye, barley and maize harvested in 1989-2001 in several regions of Russia has been conducted. A total of 5652 samples of cereals were analysed for DON and ZEN by using TLC and normal-phase HPLC with UV-detector. DON was detected in 69% of 2166 samples from Krasnodar region which is considered to be the major Fusarium endemic region of Russia. The contamination levels ranged from 0.1 till 8.6 ppm, MTEL was exceeded in 37% of these samples. The positive correlation between DON concentration and a percentage of Fusaria-damaged wheat kernels has been shown. DON occurrence and contamination levels were much lower that for wheat. Based on the results of monitoring and the data of average actual consumption of wheat products in Russia, the estimated daily intake of DON per 1 kg of body weight (EDI)was calculated. EDI varied from 0.07 ug in 1990-1991 till 1.40 ug in 1992. Although average EDI were lower than adopted tolerable daily intake (TDI, 3 ug/kg body weight) EDIs for the North-Caucasian region in some cases exceeded TDI.