赤芝子实体新三萜化合物的结构鉴定

目的　对赤芝子实体的化学成分进行研究。方法　利用各种色谱技术进行分离,采用¹HNMR ,¹³CNMR ,¹H-¹HCOSY ,HMQC和HMBC谱进行结构鉴定。结果　从赤芝子实体中分离得到3个化合物,分别鉴定为:lucidenicacidA (1) ,3β,7β-dihydroxy-4,4,14α-trimethyl-11,15-dioxo-5α-chol-8-en-24-oic acid (2 )和lucidenic acid C (3)。结论　化合物2为新化合物,命名为赤芝酸LM₁(lucidenicacidLM₁)。     

反式曲马多及氧去甲基曲马多对映体跨血脑屏障转运

目的　研究反式曲马多(trans T)及其活性代谢物氧去甲基曲马多(M₁)对映体的跨血脑屏障转运。方法大鼠ip盐酸 trans T (16.7mg·kg^-1 或5.0mg·kg^-1) 1 h后取血、脑脊液和大脑皮层,高效毛细管电泳测定血清、脑脊液和大脑皮层中 trans T和M₁ 对映体的浓度。结果　trans T和M₁ 各对映体浓度以大脑皮层中最高,血清中浓度居中,脑脊液中浓度最低。在血清中,(+)-trans T的浓度明显高于(-)-trans T的浓度,M₁ 两对映体的浓度无明显区别;在脑脊液和大脑皮层中,(+)-trans T的浓度明显高于(-)-trans T的浓度,(+)-M₁ 的浓度明显低于(-)-M₁ 的浓度。结论　trans T和M₁ 的跨血脑屏障转运具有立体选择性,脑组织中分别以(+)-对映体和(-)-对映体浓度较高。     

毛细管电泳法分离测定血浆样品中盐酸多奈哌齐对映体

目的分离、测定血浆样品中盐酸多奈哌齐对映体的含量。方法建立高效毛细管电泳法血浆中加入内标L-酒石酸布托菲诺后，碱化并以异丙醇-正己烷(3∶97)提取血浆样品，采用未涂层熔融石英毛细管柱(70 cm×50 μm ID)，以25 mmol·L^-1磷酸-三乙胺(pH 2.5)为运行缓冲液，2.5%磺化β-环糊精为手性添加剂，进样电压为-20 kV，进样时间为15 s，运行电压为-20 kV。对盐酸多奈哌齐对映体进行电泳分离，并进行家兔血浆样品中盐酸多奈哌齐对映体定量分析的方法验证。结果在上述电泳条件下，盐酸多奈哌齐对映体可达基线分离。测得血浆样品中R(-)-多奈哌齐的回归方程为：A_s/A_i=0.024 2+0.289 2C，r=0.999 2，S(+)-多奈哌齐的回归方程为：A_s/A_i=0.010 8+0.273 7C，r=0.999 7，线性范围为0.1～5 mg·L^-1，检测限为0.05 mg·L^-1。结论本法与手性柱HPLC法相比，有高效、快速和经济等优点。     

乌骨藤中C21甾体苷诱导SACC-83和SACC-LM细胞凋亡的作用及其机制研究

目的 研究乌骨藤提取物C₂₁甾体苷对唾液腺腺样囊性癌（salivary adenoid cystic carcinomacv,SACC）低侵袭细胞株（salivary adenoid cystic carcinoma-83,SACC-83）和肺高转移细胞株（SACC-LM）增殖抑制和诱导凋亡的作用及其机制。方法 用不同浓度（5,10,20,40,60,80,100 μmol·L^-1） C₂₁甾体苷处理SACC-83和SACC-LM细胞48 h后,MTT法检测细胞活力,并计算药物的IC₂₀和IC₅₀;细胞克隆形成实验检测细胞增殖能力;流式细胞术检测SACC-83及SACC-LM细胞凋亡情况;实时荧光定量PCR和Western blot检测SACC-83和SACC-LM细胞Bcl-2、Bax、caspase 3的mRNA和蛋白的表达。结果 不同浓度的C₂₁甾体苷降低SACC-83和SACC-LM细胞活力,抑制细胞增殖,并且作用于SACC-83细胞C₂₁甾体苷的IC₂₀浓度为7.49 μmol·L^-1,IC₅₀浓度为38.34 μmol·L^-1;作用于SACC-LM细胞的C₂₁甾体苷IC₂₀浓度为9.30 μmol·L^-1,IC₅₀浓度为46.04 μmol·L^-1;细胞克隆集落形成明显减少。C₂₁甾体苷IC₂₀浓度分别促进SACC-83及SACC-LM细胞凋亡,且随着给药时间延长,凋亡率增加,具有显著性差异（P<0.05,P<0.01）。经7.49,9.30 μmol·L^-1 C₂₁甾体苷分别处理SACC-83及SACC-LM细胞后,Bcl-2的mRNA及蛋白水平显著降低（P<0.01）,而Bax、Caspase 3的mRNA和蛋白水平显著升高（P<0.05）。结论 乌骨藤C₂₁甾体苷抑制SACC-83及SACC-LM细胞增殖、促进凋亡,其作用机制可能与调控Bcl-2、Bax和Caspase 3表达有关。     

液相色谱-串联质谱法测定犬血浆中布地奈德

建立液相色谱-串联质谱法测定犬血浆中布地奈德。血浆样品碱化后，经乙酸乙酯液-液萃取，以乙腈-5 mmol·L^-1醋酸铵(60∶40，v/v)为流动相，Capcell Pak C₁₈ MG柱分离；采用电喷雾电离源，以多反应监测(MRM)方式进行负离子检测，用于定量分析的离子反应分别为m/z 489→m/z 357(布地奈德)和m/z 493→m/z 413(内标，曲安奈德)。测定血浆中布地奈德方法的线性范围为25.0～2 000 pg·mL^-1，定量下限为25.0 pg·mL^-1，日内、日间精密度(RSD)均小于15%，准确度(RE)在-8.1%～-1.7%。应用本法研究6只比格犬单次和多次给予布地奈德缓释胶囊9 mg后的药代动力学结果显示：单次给药后T_max为(3.5±3.3) h，C_max为(786±498) pg·mL^-1；多次给药后C_max为(2 142±1 515) pg·mL^-1。该法选择性强、灵敏度高、操作简便，适用于布地奈德缓释制剂的药代动力学研究。     

温莪术挥发油中莪术醇的含量测定

本文报告用薄层层析-红外光谱法测定莪术挥发油中莪术醇的含量。以石油醚-丙酮(96:4)为展开剂,在硅胶G板上将挥发油中极性较小的干扰物质层析至高R_f值区而将莪术醇留在原点附近,然后无需标记出莪术醇的斑点,根据经实验求出的莪术醇的R_f值直接将薄层上莪术醇部分刮下洗脱,再以红外光谱中莪术醇分子中亚乙烯基的吸收峰(面外弯曲振动890 cm^-1,11.24μm)为定量谱带,用四氯化碳为溶剂,以红外光谱法测定莪术醇的含量。由于亚乙烯基谱带的专属性较强,除干扰组分外,其他组分与莪术醇无需严格分离。本法操作简便,结果稳定,统计结果表明,8次测定的平均相对偏差小于2%。     

液相色谱-质谱-质谱联用法测定人血浆中氯诺昔康液相色谱-质谱-质谱联用法测定人血浆中氯诺昔康

目的建立测定人血浆中氯诺昔康的液相色谱-质谱-质谱联用法,并用于中国受试者口服氯诺昔康的体内药代动力学研究。方法血浆样品经液-液萃取后,以甲醇-水-甲酸(80∶20∶0.5)为流动相,吡罗昔康为内标,采用Zorbax XDB-C8柱分离,通过液相色谱串联质谱,以选择反应监测(SRM)方式进行检测。定量分析离子反应分别为m/z 372→121(氯诺昔康)和m/z 332→121(吡罗昔康)。结果线性范围为2.0～1 600 μg·L^-1,定量下限为2.0 μg·L^-1。18名受试者po氯诺昔康8 mg后主要药动学参数t_1/2为(4.7±1.1) h,AUC_0-∞为(5.5±2.4) mg·h·L^-1。而另一名受试者t_1/2为105 h,AUC_0-∞为189.5 mg·h·L^-1。结论该法灵敏度高,线性范围宽,操作简便、快速,适用于临床药代动力学研究。     

柱前衍生-高效液相色谱荧光法测定大鼠血浆中硫普罗宁

建立了测定大鼠血浆中硫普罗宁含量的柱前衍生-高效液相色谱荧光法。血浆样品经二硫苏糖醇(DTT)还原后，再经N-(1-芘)马来酰亚胺(NPM)衍生化后进样测定。以0.2%乙酸(含0.015 mol·L^-1磷酸二氢钾)-乙腈(56∶44)为流动相，采用Kromasil C₁₈色谱柱分离，在λ_ex=340 nm，λ_em=375 nm下荧光检测。硫普罗宁线性范围为0.1～10.0 μg·mL^-1，定量下限为0.1 mg·L^-1，日内、日间精密度(RSD)分别为5.3%～10.8%和7.0%～10.8%，提取回收率在73.7%～79.7%。大鼠单剂量给予硫普罗宁后，药代动力学行为符合二室模型。该法准确可靠，专属性强，适用于硫普罗宁的药代动力学研究。     

两种测定小鼠体内力达霉素药代动力学方法的比较

目的对测定小鼠血清浓度的总放射性方法和经过高效液相色谱分离后再测定放射性的两种方法进行比较。方法首先对力达霉素进行同位素标记并且分离纯化。给小鼠尾iv ¹²⁵I-力达霉素(100 μg·kg^-1)，采集血样，样品经两种方法测定。对得到的药代动力学参数进行统计学处理。结果两种方法测定得到的药代动力学参数(V_d, t_1/2α, t_1/2β, k₂₁, k₁₀, k₁₂, AUC和CL) 存在显著性的差异 (P<0.05)。结论采用色谱分离后再测定¹²⁵I-力达霉素原形的放射性较为合理和准确。     

麻黃揮发油的研究

本文报告由麻黄中提取出一种淡黄色具有花香的挥发油,n_D²⁰1.4940,[α]_D²⁰—31.44°,d₂₀²⁰0.8819。将此油反复减压分馏和用氧化铝柱层离后得到一种萜醇(C₁₀H₁₈O),经制备其衍生物和红外吸收光谱、气相层离法鉴定,证明为l-a-萜松醇。     

手性溶液萃取分离氧氟沙星对映体

目的研究氧氟沙星对映体在手性环境中的萃取分配行为,为外消旋体膜萃取拆分提供理论和设计依据。方法以0.25 mol·L^-1 L-二苯甲酰酒石酸醇溶液为有机相,含0.14 g·L^-1氧氟沙星的0.1 mol·L^-1磷酸氢二钠/磷酸缓冲液为水相,考察了pH对氧氟沙星对映体在水-有机相分配系数(K)和分离因子(α)的影响,研究了不同碳数醇的溶剂化效应,同时运用中空纤维膜对氧氟沙星外消旋体进行初步分离。结果以癸醇为溶剂,pH 6.86时,L-二苯甲酰酒石酸对氧氟沙星外消旋体萃取分配系数大于4.7,α达1.18;用中空纤维支载液膜双有机相逆流分级萃取技术,11个22 cm长膜器串联,产品光学纯度达90%以上。结论L-二苯甲酰酒石酸在醇溶剂中对磷酸氢二钠缓冲液中的RS-氧氟沙星对映体有较强的立体选择性,其中癸醇的效果最好,pH对K和α有较大影响,综合K和α值,取pH 6.86时,手性萃取效果最佳;利用高效中空纤维膜,可使RS氧氟沙星对映体得到不同程度的分离。     

INFLUENCE OF DIETARY RESTRICTION AND PROTEIN DEFICIENCY ON PLASMA HALF-LIFE AND TISSUE DISTRIBUTION OF TETRACYCLINE IN RATS

The effects of dietary restriction and protein deficiency on plasma half-life and tissue distribution of tetracycline were studied in rats by feeding either a 20% protein diet in restricted quantity or a 9% protein diet ad lib and compared with rats given a 20% protein diet ad lib (control group). 1 It was observed that half-life of tetracycline was shortened and that plasma and tissue C_min levels at steady-state were lower in undernourished rats. Tissue concentrations in liver, kidney, muscle and bone correlated well with plasma levels. A high degree of correlation was also observed between plasma and tonsillar concentrations of tetracycline in human subjects. 2 These studies indicate that undernourished subjects may require an altered dosage regimen of tetracycline to maintain effective steady-state concentrations of the drug.     

何首乌有效成分二苯乙烯苷的药代动力学研究

目的建立小鼠和兔血浆中二苯乙烯苷浓度的HPLC测定方法,研究何首乌中二苯乙烯苷在小鼠和兔体内的药代动力学行为。方法用Diamonsil^TM C₁₈色谱柱(250 mm×4.6 mm,5 μm),以乙腈-甲醇-1%甲酸(15∶18∶67)为流动相,流速1.0 mL·min^-1,检测波长320 nm。结果线性范围为0.41～42.0 μg·mL^-1(γ=0.9999),最低检测浓度为0.051 μg·mL^-1。高、中、低3种不同浓度的平均回收率分别为97.98%,101.7%和104.5%,日内精密度RSD分别为8.7%,2.9%和5.5%。小鼠和兔iv二苯乙烯苷后药代动力学行为均符合二室模型,药代动力学参数分别为:T_1/2α=1.9,2.7 min;T_1/2β=8.3,13.5 min;K₂₁=6.6,4.2 h^-1;K₁₂=3.8,3.0 h^-1;K₁₀=16.0,11.2 h^-1;V_c=0.090,0.198 L·kg^-1;AUC=6.918,4.530 mg·h·L^-1;CL=1.445,2.208 L·h^-1·kg^-1。小鼠ig给药后二苯乙烯苷在胃肠道内的吸收不规则,且血药浓度很低,药代动力学行为不符合房室模型。结论建立了二苯乙烯苷血药浓度的HPLC测定方法,阐明了二苯乙烯苷的药代动力学特征。方法的专属性高,操作简便,结果准确。     

姜黄及其制剂中姜黄素类化合物的高效液相色谱分离与测定

姜黄为姜料植物Curcuma longa L.的根茎,为一常用中药。近年来用于治疗高脂血症有较好疗效,且无毒副作用。其主要化学成分为姜黄素、去甲氧基姜黄素及二去甲氧基姜黄素。实验证实姜黄素为其降血脂的主要有效成分。姜黄素类化合物过去多采用在420 nm附近测定黄色素的方法,此类方法不够稳定,易受其它黄色素的干扰。利用姜黄素与硼生成红色络合物而进行测定,比较专一而灵敏。对     

Genotoxicity of tetracycline as an emerging pollutant on root meristem cells of wheat (Triticum aestivum L.)

Increasing attention has been paid to antibiotic contamination as an increasingly serious environmental issue. Tetracycline has been widely used for decades in human and veterinary medicines, with incremental residues in the environment and adverse influences on living organisms. In the present study, the genetic toxicity of tetracycline was investigated using a bioassay method with wheat (Triticum aestivum L.) root‐meristem cells at a concentration range of 0.25–300 mg L^?1 and exposure times of 24, 48, and 72 h. The results indicated that tetracycline at lower concentrations (0.25–1 mg L^?1) stimulated cell mitotic division, whereas at 50–300 mg L^?1 concentration caused a concentration‐related decrease in mitotic index (MI). The lower tetracycline concentrations induced a slight increase in the frequency of micronucleus (MN), chromosomal aberration (CA), and sister chromatid exchange (SCE) in wheat root tips. However, there were significant increases in these indices at higher concentrations in concentration‐ and time‐dependent manners, including the frequencies of MN (25–200 mg L^?1), CA (10–200 mg L^?1), and SCE (5–200 mg L^?1), respectively. The inducement of MN, CA, and SCE decreased at 250 and 300 mg L^?1 due to acute cell toxicity for all tested times. Comparatively, SCE was the most sensitive, followed by CA, with MN the least sensitive to the genotoxicity of tetracycline in wheat. These results imply that tetracycline may be genotoxic to plant cells, and exposure to tetracycline may pose a genotoxic risk to living organisms. The results also suggest that the wheat bioassay was efficient, simple, and reproducible in monitoring the genotoxicity of tetracycline in the environment. © 2010 Wiley Periodicals, Inc. Environ Toxicol, 2011.     

绵毛马兜铃化学成分研究——Ⅳ.绵毛马兜铃内酯的化学结构测定

从马兜铃属植物绵毛马兜铃Aristolochia mollissima Hance根茎中分得9种(Ⅰ～Ⅸ)成分,其中结晶Ⅲ为一新倍半萜内酯,用光谱法测定了化学结构,命名为绵毛马兜铃内酯Mollislactone,晶Ⅰ为尿囊素,晶Ⅱ为马兜铃内酯,晶Ⅳ为β-谷甾醇,晶Ⅸ为马兜铃酸,其余成分尚在研究中。     

一叶萩碱对5种神经递质受体和蛙脊髓膜电位的影响

Using radioligand binding assay method, securinine was shown to be specifically bound to GABA receptors of rat brain with IC₅₀ of 10^(-4)～10^-5mol/L. However, securinine did not exhibit affinity for a₁-, a₂- adrenoceptors and M-cholinergic receptors of rat brain and the β-adrenoceptors of duck erythrocyte membrane. In isolated and perfused bullfrog spinal cord experiment, the depolarization induced by GABA was slightly depressed by securinine (1 mmol/L), but no antagonistic effect on glycine or taurine evoked depolarization was observed with securinine. These results indicate that securinine is a weak GABA antagonist.     

Promotion of IL-8 production in PMA-stimulated HL-60 cells by sesquiterpene quinones from a marine sponge, <Emphasis Type="Italic">Hippospongia</Emphasis> sp.

An extract of a marine sponge, Hippospongia sp., collected in Palau has inhibitory activity against colony formation by Chinese hamster V79 cells. Bioassay-guided isolation gave eleven sesquiterpene quinones. Compounds 1–8 inhibited colony formation by V79 cells with EC₅₀ values between 0.6 and 2.8 μmol L⁻¹. Their effects on the production of an inflammatory cytokine, IL-8, in tetradecanoyl phorbol acetate (PMA)-stimulated HL-60 cells were also investigated, because IL-8 production is sometimes correlated with inhibition of cell growth. Ilimaquinone (1) and its 5 epimer (2) had similar activity against V79 cells (EC₅₀ = 2.8 and 2.3 μmol L⁻¹, respectively) but did not modulate IL-8 production even at 10 μmol L⁻¹. Smenospongidine (3) and its 5 epimer (4), smenospongiarine (5) and its 5 epimer (6), and smenospongine (7) and its 5-epimer (8), at 10 μmol L⁻¹, promoted IL-8 production. Compounds 3, 5, and 7 had slightly stronger activity against V79 cells (EC₅₀ = 0.6, 1.7, and 0.8 μmol L⁻¹, respectively) than the corresponding 5 epimers 4, 6, and 8 (EC₅₀ = 0.8, 2.3, and 2.0 μmol L⁻¹, respectively). A similar structure–activity relationship was observed for promotion of IL-8 production. This is the first report of modulation of IL-8 production by these sesquiterpene quinones.     

核磁共振氢谱鉴别植物中药的研究

目的：建立核磁共振氢谱法鉴别植物中药的理论和方法。方法：选择适当的提取分离工艺,制备植物中药的特征总提物,在NMR波谱仪上测定中药的1HNMR指纹图;对特征总提物进一步分离,得到单体化合物后,通过结构鉴定和NMR研究,以对指纹图中的各特征谱线进行归属。结果：所研究的各种植物中药的1HNMR指纹图均有高度的特征性和重现性,同一品种不同产地的样品的¹HNMR指纹图有很好的一致性;人参、天麻、黄连等多数植物中药特征总提物的¹HNMR指纹图主要显示了其活性成分的特征共振峰,人参的¹HNMR指纹图在δ 0.87～1.64区及δ 4.16～5.12区的共振信号能准确地反映人参皂苷的存在,天麻在δ 6.60～7.35区显示出其活性成分天麻苷和对-羟基苯甲醇的存在,而黄连的1HNMR指纹图主要为原小檗碱型生物碱的贡献。结论：用本文试验方法测定的多数植物中药特征总提物的¹HNMR指纹图及其数据是这些中药的特征共振信号和数据,可作为鉴别植物中药的参照及其相对标准图谱。     

Identification and characterization of flavonoids from semen zizyphi spinosae by high-performance liquid chromatography/linear ion trap FTICR hybrid mass spectrometry

Semen zizyphi spinosae (SZS) has been used to treat insomnia and anxiety for thousands of years. In this paper, a novel high-performance liquid chromatography coupled with the photodiode array detector/linear ion trap-MS ⁿ (HPLC-PDA/LTQ-MS ⁿ ) method was established to separate and identify flavonoids from the extract of SZS. Separation was performed on an HYPERSIL C₁₈ column by gradient elution using CH₃CN/H₂O–CH₃COOH as the mobile phase at a flow rate of 0.8 ml/min. UV spectral data, accurate molecular weights, and multi-stage MS/MS fragmentation information were obtained. Electrospray ionization/MS/MS fragmentation patterns were proposed. Nineteen flavonoid glycosides were identified or tentatively characterized based on their retention time, UV spectral data, accurate molecular weights, and mass fragmentation behavior. The method was useful for separation and identification of the flavonoid components from SZS and could be applied to other complex samples, especially for minor constituents.