气相色谱—质谱法测定川芎哚血药浓度

目的 :建立测定大鼠血浆样品中川芎哚浓度的气相色谱 -质谱 (GC -MS)定量分析方法。方法 :血浆样品经乙醚 -异丙醇 (10∶1)提取、硅烷化处理后 ,选择GC -MS选择性离子和辅助定量离子监测。结果 :本法线性范围为 2～ 6 0ng·mL-1,灵敏度为 1ng·mL-1,回收率达 82 % ,日内日间RSD均小于 9 0 %。结论 :本法为川芎哚临床药代动力学研究提供了准确、可靠的方法。     

川芎哚及其类似物对凝血功能和血液流变学的影响

川芎哚 (川芎Ⅲ号碱 ,ｐｅｒｌｏｌｙｒｉｎｅ,ＰＬ)是中药川芎的有效活性成分之一[1] ,初步药理实验表明 ,川芎哚对冠心病有一定的疗效 ,但它在动物体内代谢迅速 ,排泄较快 ,生物利用度低及药效维持时间较短[2 ] ,因此有必要对其进行结构改造和药理筛选研究[2 ] 。本文对合成的川芎哚及其类似物进行了体外凝血实验及血液流变学实验研究。1　材料　　川芎嗪盐酸盐 (ＴＭＰ)、川芎哚 (ＰＬ)、甲磺酸川芎哚Ｔ 1)、阿魏酸川芎哚 (Ｔ 2 )、阿魏酸哌嗪 (Ｔ 3)、阿魏酸川芎嗪 (Ｔ 4 )以及川芎哚的类似物Ｔ 5～Ｔ 17,均为本实验室制备[2 ,3 ] ;钙试…     

稳定同位素结合GC-MS法测定大鼠体内川芎哚的药物动力学参数(英文)

目的:测定川芎哚(川芎Ⅲ号碱,perlolyrine)的药动学参数。方法:以[2-~(15)N]川芎哚为内标准及GC-MS的SIM(选择性离子监测)为检测手段,定量测定大鼠体内川芎哚的含量及其药代动力学参数。结果:大鼠灌胃给予川芎哚2mg·kg~(-1)后,川芎哚在大鼠体内呈二室模型分布,其药代动力学参数为:T_((1/2)α)=0.33h,T_((1/2)β)=4.52h,T_(1/2)(ka)=0.14h,T_(max)=0.35h,C_(max)=18.84μg/L,K_(12)=0.88h~(-1),K_(21)=0.42h~(-1),K_(10)=0.32h~(-1),V/F=109.22 L·kg~(-1),AUC=112.68μg·h·L~(-1)。结论:本法灵敏度高、特异性强且准确性好,为测定川草哚药代动力学参数提供了实用的分析方法。本研究为川芎哚临床应用提供了重要的参考资料。     

川芎哚药代动力学参数的稳定同位素法测定

目的　测定川芎哚 (川芎III号碱 ,perlolyrine)的药动学参数。方法　以氘标川芎哚为内标准及GC MS的SIM(选择性离子监测 )为检测手段 ,定量测定大鼠体内川芎哚的含量及其药代动力学参数。结果　川芎哚在大鼠体内呈二室模型分布 ,其药代动力学参数为 :T1/ 2α=0 34h ,T1/ 2 β=4 5 9h ,T1/ 2 (Ka) =0 12h ,Tmax=0 36h ,Cmax=18 86μg·L-1,K12 =0 87h-1,K2 1=0 40h-1,K10 =0 31h-1,VB=10 7 86L·kg-1,AUC =99 6 8μg·h·L-1。结论　本法灵敏度高、特异性强和准确性好 ,为临床应用提供了科学依据。     

川芎哚及其类似物对血小板聚集和实验性血栓形成的影响

目的　研究川芎哚 (T 1)及其类似物 (T 2～T 17)对血小板聚集和实验性血栓形成的影响。方法　T 1～T 17进行体外血小板聚集实验 ,计算血小板聚集抑制率 ,从中选出T 2、T 3、T 4、T 5、T 6、T 8、T 11、T 12及T 14按Umet su氏法进行实验性血栓形成实验 ,计算血栓形成抑制率 ,并与川芎嗪 (TMP)进行比较。结果　所有合成药物均有一定程度的抗血小板作用 ,其中T 5和T 6的抗血小板作用强于TMP ;各试验药物均有一定程度的抗血栓作用 ,其中T 5、T 6、T 11、T 12及T 14抗血栓作用时间和强度明显大于TMP。结论　川芎哚及其类似物具有抗血小板和抗血栓作用 ,其中T 5和T 6的作用强于TMP ,但川芎哚的作用弱于TMP。     

川芎哚的结构改造及生物活性研究

目的:寻找新型的心血管药物。方法:根据推理药物设计原理,设计合成目标化合物,并研究其生物活性。结果:合成了16个川芎哚的类似物(I～XVI),其中8个化合物(I,I,V～VI,IX,XI和XV)是新化合物。结论:所合成的川芎哚类似物均有一定程度的抗血小板聚集作用,其中I～IV,VI,X,XI及XII有一定程度的抗血栓作用。     

罂粟碱的体内与体外代谢物研究

利用HPLC-MSⁿ检测罂粟碱及其在大鼠体内(大鼠粪便)和体外(肝微粒体, 肠道菌)代谢物。体内与体外的代谢物经C₁₈小柱进行富集和纯化后, 直接采用优化的HPLC-ESI/ITMSⁿ方法对样品进样分析。以罂粟碱标准品为对象优化高效液相色谱/电喷雾离子阱质谱(HPLC-ESI/ITMSⁿ )实验条件, 分析总结其电喷雾质谱的一级电离规律和多级质谱裂解规律, 作为罂粟碱大鼠体内与体外代谢物结构分析的依据。代谢物的结构推导主要依据代谢物的色谱保留时间及其电喷雾离子阱质谱HPLC-ESI/ITMSⁿ电离规律。在大鼠粪便中有原药及其8种代谢产物。体外代谢物检测到原药的脱甲基和羟基化。     

川芎哚及其类似物的抗血小板与抗血栓作用

为了研究川芎哚 (PL)及其类似物 (T 1～T 17)的抗血小板和抗血栓作用 ,T 1～T 17按Foye法进行体外血小板聚集实验 ,计算血小板聚集抑制率 ,从中选出T 1,T 2 ,T 3,T 4 ,T 6 ,T 8,T 11,T 12及T 14按Chandler法进行实验性血栓形成实验 ,计算血栓形成抑制率 ,并与川芎嗪 (Lig)进行比较 .结果表明 :所有合成药物均有一定程度的抗血小板作用 ,其中T 4和T 6的抗血小板作用强于Lig ;各试验药物均有一定程度的抗血栓作用 ,其中T 4 ,T 6 ,T 11,T 12及T 14抗血栓作用强度大于Lig .由此可见 ,PL及其类似物具有抗血小板和抗血栓作用 ,其中T 4和T 6的作用强于Lig ,但PL的作用弱于Lig     

LC/MS分析大鼠体内氧化苦参碱及其主要代谢物LC/MS分析大鼠体内氧化苦参碱及其主要代谢物

目的研究氧化苦参碱在大鼠体内的主要代谢产物。方法以氧化苦参碱和苦参碱为对象优化液相色谱/电喷雾离子阱质谱(LC/ESI-ITMSⁿ)实验条件，分析总结其电喷雾质谱的一级电离规律和二级质谱裂解规律，作为氧化苦参碱大鼠体内代谢物结构分析的依据。健康大鼠腹腔肌注40 mg·kg^-1氧化苦参碱，收集0~24 h的尿样，尿样中的代谢物经C₁₈小柱进行富集与纯化后，在优化的LC/ESI-ITMSⁿ条件下进样分析。代谢物的结构推导主要依据代谢物的色谱保留时间及其电喷雾离子阱质谱(ESI-ITMSⁿ)电离规律。结果在大鼠尿样中有原药及其6种I相氧化及还原代谢产物，且主要代谢物为苦参碱。未发现II相代谢物。结论本法不仅操作简便、快速，而且灵敏度高、专属性强。该分析技术是研究药物代谢最有效的方法之一。     

川芎中川芎嗪和阿魏酸含量的毛细管电泳测定

目的：研究川芎中川芎嗪和阿魏酸的毛细管电泳分离分析的可行性。方法：通过用毛细管区带电泳分离、紫外检测模式研究内标、电泳缓冲液、进样方式、分离电压等对样品中川芎嗪和阿魏酸分离与定量的影响,优化实验条件。结果：以对硝基苯甲酸为内标,未涂层熔融石英毛细管(39.5 cm×50 μm ID,有效分离长度34.8 cm)为分离通道,30 mmol.L^-1硼砂缓冲液(pH 9.43)为电泳介质,34 kPa.s压力进样,17 kV分离电压,295 nm检测,川芎嗪和阿魏酸分别在7～423 μg.mL^-1和4～900 μg.mL^-1范围内可进行定量检测,回收率分别为100.9％±1.9％和99.8％±1.0％。结论：毛细管电泳法可用于川芎中川芎嗪和阿魏酸的同时分离分析。     

丁基苯酞的体内代谢转化研究

大鼠po丁基苯酞(NBP),定时收集尿液,经酶水解、提取浓缩、衍生化处理后用GC/MS分析。在给药后0～24h及24～48h尿中,NBP原药含量很低,主要以代谢物形式存在,依次为γ-羟基、β-羟基与3-羟基取代物。NBP体内代谢结果与肝微粒体中代谢结果基本一致。     

Isolation and antihyperglycemic activity of flavonoid from flower petals of <Emphasis Type="Italic">Opuntia stricta</Emphasis>

The fresh flowers of Opuntia stricta were collected and extracted with 85% methanol under reflux. The concentrated extract was fractionated with Et₂O, EtOAc and petroleum ether. Rhamnetin 3-O-rutinoside was obtained from EtOAc fraction, as confirmed through ¹H and ¹³C NMR, ¹H-¹H NOESY, ¹H-¹³C HSQC, and mass spectra. The isolated flavonoid glycoside was investigated for its antihyperglycemic activity on alloxan-induced diabetic male albino rats.     

Amino acid sequence of a neutral phospholipase A2 (III) in the venom of Bungarus fasciatus

Two phospholipases were found in the venom of Bungarus fasciatus, one in fraction III, the other in fraction X of the chromatographic separation. A neutral PLA₂(III) purified from fraction III was subjected to amino acid sequencing by means of an automated sequenator applied to the intact RCM-PLA₂ (III) and the individual peptides obtained from HPLC separation of the three types of enzymatic peptides. PLA(III) was shown to consist of 118 amino acid residues with 14 half-cystines. It is 65% homologous to the basic PLA₂ obtained from fraction X.     

Effect of pH on uptake,tissue distribution and retention of hexavalent chromium in rainbow trout (Salmo gairdneri)

Uptake, distribution and retention of chromium in rainbow trout (Salmo guirdneri) was studied after short-term (2–4 days) exposure to ⁵¹CrO₄^2? -containing Na₂CrO₄ solutions of different concentrations (2–50 mg/l Cr) and pH (7.8 and 6.5). At pH 7.8, highest contents of chromium were found in gill, liver, kidney, and digestive tract of the trout. Chromium was not distributed evenly among the different subcellular fractions of the tissues, but was concentrated in the nuclear fraction of the gill tissue and in the soluble fraction of the kidney and liver tissue. Upon transfer of exposed fish to tap-water, chromium was rapidly eliminated from blood, gill and digestive tract. However, chromium contents tended to remain high in kidney and liver. When the pH was decreased from 7.8 to 6.5, the lethal action of hexavalent chromium increased and a different pattern of accumulation and elimination of chromium was observed. The major differences were found in the gills, which concentrated significantly more chromium at pH 6.5 than at pH 7.8, irrespective of the exposure time and concentration. As an electronspin-resonance signal characteristic for trivalent or pentavalent chromium was detected in the gills, the differences must have been at least partly due to the higher oxidizing action of hexavalent chromium at the lower pH.     

Isotopically labeled boldenone as a better marker of derivatization efficiency for improved quality control in anti‐doping analysis

At present, anti‐doping laboratories use androsterone, a major urinary steroid metabolite, to evaluate completeness of the derivatization step. This is typically done by calculating the ratio of mono‐trimethylsilyl (TMS) androsterone to the total mono‐ and di‐TMS androsterone. Certain samples may show an elevated percentage of mono‐TMS androsterone indicating a failed derivatization step. In such cases, the laboratory would have to repeat the analysis or perform other remedial actions to ensure that completeness of derivatization is achieved. We have noticed that a poorly derivatized positive control sample spiked with various target analytes has a disproportionally low abundance of the di‐TMS derivatives of boldenone and 18‐nor‐17β‐hydroxymethyl‐17α‐methylandrosta‐1,4,13‐trien‐3‐one (methandienone long‐term metabolite). A follow‐up investigation confirmed that 1,4‐diene‐3‐one steroids are more likely to fail during the trimethylsilylation step. To better control derivatization efficiency, ¹³C₃‐boldenone (13C‐BLD) was incorporated into our routine procedure as an additional internal standard. Analysis of a large number of urine samples has shown that derivatization of 13C‐BLD could be grossly incomplete even in cases when mono‐TMS androsterone is well below 1%. In other words, one or both of boldenone and the long‐term metabolite of methandienone could remain undetected unless the laboratory has the means to recognize samples where derivatization of 1,4‐diene‐3‐one steroids failed.     

Studies on the alkaloidal components of the fruits ofLycium chinense

N₉-Formylharman, 1-carbomethoxy-β-carboline and perlolyrine with an unidentified alkaloid (C₁₀H₁₃NO₂, M⁺ 179.094) have been isolated from the fruits ofLycium chinense Miller (Solanaceae) and characterized on the basis of chemical and physical evidence.     

Opioidergic mechanisms underlying the actions of Vitex agnus-castus L.

Vitex agnus-castus (VAC) has been used since ancient Greek times and has been shown clinically to be effective for the treatment of pre-menstrual syndrome. However, its mechanism of action has only been partially determined. Compounds, fractions, and extracts isolated from VAC were used in this study to thoroughly investigate possible opioidergic activity. First, an extract of VAC was found to bind and activate μ- and δ-, but not κ-opioid receptor subtypes (MOR, DOR, and KOR respectively). The extract was then resuspended in 10% methanol and partitioned sequentially with petroleum ether, CHCl₃, and EtOAc to form four fractions including a water fraction. The highest affinity for MOR was concentrated in the CHCl₃ fraction, whereas the highest affinity for DOR was found in the CHCl₃ and EtOAc fractions. The petroleum ether fraction had the highest agonist activity at MOR and DOR. Several flavonoids from VAC were found to bind to both MOR and DOR in a dose-dependent manner; however only casticin, a marker compound for genus Vitex, was found to have agonist activity selective for DOR at high concentrations. These results suggest VAC may exert its therapeutic effects through the activation of MOR, DOR, but not KOR.     

Simultaneous HPLC analysis of ceramide and dihydroceramide in human hairs

Ceramide, a major class of hair lipid, can help determine the physicochemical properties of human hairs such as the chemical diffusion barrier and water retention. In this study, we developed a quantitation method for ceramide and dihydroceramide, a saturated form of ceramide, in human hairs. Lipids were extracted with ethanol from human hairs spiked with N-oleoyl-D-erythro-C₁₇ sphingosine, an internal standard. Ceramide and dihydroceramide were resolved by TLC and deacylated by sphingolipid-ceramide deacylase to release sphingosine and dihydrosphingosine, respectively. The hair content of ceramide was measured by HPLC following derivatization with o-phthalaldehyde. The limits of detection and quantification for ceramide extracted from hair fibers were 0.1 and 1 pmol, respectively. The linear range of hair weight for determining ceramide and dihydroceramide contents was 1 to 50 mg, with R² values of 0.9695 and 0.9898, respectively. The recoveries of ceramide and dihydroceramide from intra-day and interday assays were 99.55% to 98.53%, respectively. Concentrations of dihydroceramide from the hair roots to distal tip ends ranged from 10.42 ± 2.19 to 1.20 ± 0.11 nmol/g hair, while those of ceramide ranged from 2.27 ± 0.22 to 1.47 ± 0.15 nmol/g hair. The present analytical method provides a simultaneous and reproducible quantification of ceramide and dihydroceramide, and may be used as a potential biomarker for lipid abnormality-related diseases.