不同炮制方法对中药延胡索中有效成分含量的影响

目的:比较醋炙、醋煮、酒炙、醋烘等炮制方法对延胡索中有效成分含量的影响.方法:将延胡索生品分别经醋炙、醋煮、醋烘、酒炙后,以水超声法对不同炮制品提取,高效液相色谱法测定去氢紫堇碱、延胡索乙素成分的含量,乙腈:0.1%磷酸溶液=72:28作为试验的流动相,色谱柱AgilentC18色谱柱,柱温为室温,检测波长为282nm.结果:延胡索经不同炮制后生物碱去氢紫堇碱含量均有所升高,醋烘、醋炙、酒炙延胡索中延胡索乙素的含量有所上升,差异有统计学意义(P<0.05).结论:高效液相色谱法测定中药延胡索不同炮制品中的2种有效成分,操作步骤简单、重复性好,根据药材的治疗需要,采用醋烘、醋炙、酒炙等炮制方法可以增加延胡索有效成分的含量.     

HPLC法测定黄柏炮制前后盐酸小檗碱的含量

目的考察黄柏炮制前后盐酸小檗碱的含量变化。方法色谱柱:Lana C18柱(4.6mm×250mm,5μm,美国Phenom enex公司):流动相:甲醇-乙腈-水(42∶16∶42)(含0.05mol.L-1十二烷基硫酸钠及0.037mol.L-1酒石酸);检测波长:345nm。结果盐酸小檗碱在0.1108~1.108μg范围内,浓度与峰面积线性关系良好(r=0.9999),样品平均回收率为99.12%,RSD为1.62%。3批黄柏经过炮制后其含有盐酸小檗碱的含量均降低。结论该方法简便、快速准确、灵敏度高,重现性好。有助于为黄柏合理的炮制工艺提供依据,可用于其炮制品的质量控制。     

炮制对黄柏中3种生物碱含量的影响

目的:考察炮制对黄柏中3种生物碱含量的影响。方法:选用高效液相色谱(HPLC)法测定3种炮制(酒炙、盐炙和炒炭)品中3种黄柏生物碱的含量。色谱柱为迪马C18(250mm×4.6mm,5μm),流动相为乙腈-0.1%磷酸(48∶52,每100mL含十二烷基磺酸钠0.1g),检测波长为265nm,流速为1.0mL·min-1,柱温为室温,进样量为10μL。结果:盐酸小檗碱、盐酸巴马汀和盐酸药根碱含量在炮制后普遍降低,以炒炭最为显著;也有个别特例。结论:黄柏在炮制后(酒炙品、盐炙品)仍然可以选择3种生物碱作为质量控制的指标。本方法简单、快速、准确,适用于同时测定黄柏炮制品中3种生物碱的含量。     

黄柏炮制过程中生物碱成分含量变化的研究

目的探究黄柏炮制过程中生物碱成分的量变和质变。方法采用高效液相色谱法测定黄柏不同炮制品,在不同温度下生物碱含量的变化。结果黄柏经炮制之后,随着温度的增加,其原有的生物碱、小檗碱、黄柏碱含量降低,并会生成新的化学成分小檗红碱,其含量随着温度的升高而增加。结论黄柏炮制过程中生物碱发生了量变与质变,加热是关键的因素,可以促进黄柏生物碱的含量变化或转化为新的化学物质。     

盐炙对关黄柏化学成分溶出的影响

目的考察盐炙对关黄柏水煎液中主要生物碱类及柠檬苦素类成分溶出的影响。方法采用HPLC法比较用分析纯NaCl、MgCl2、KCl和食盐炮制的关黄柏样品中3种主要生物碱小檗碱、药根碱、巴马汀的峰面积。色谱柱:Diamonsil ODS C18柱(250 mm×4.6 mm,5μm),流动相:乙腈(A)-体积分数为0.1%的磷酸溶液(B)(梯度洗脱程序:0～20 min:90%B～65%B,20～35 min:65%B～50%B),流速:0.8 mL.min-1,检测波长:345 nm。同时采用分光光度法测定水煎后药渣中柠檬苦素类成分的含量。结果不同种类的盐炮制的关黄柏中生物碱类成分的峰面积均有变化。药渣中的柠檬苦素成分含量也均有变化。结论盐炙对关黄柏中主要成分的水中溶出有较大影响,为阐明盐炙的炮制机理提供一定的依据。     

优选延胡索炮制方法

目的比较延胡索不同炮制工艺有效成分的含量,确定最佳的炮制工艺。方法采用HPLC测定延胡索不同炮制品中原阿片碱、盐酸小檗碱和延胡索乙素的含量,采用电位滴定法测定其炮制品中总生物碱的含量,比较不同炮制品有效成分的含量。结果延胡索生品及炮制品的醇提液中3种生物碱的含量均高于水提液,与生品比较,炮制品的水提液中3种生物碱的含量均有不同程度的升高,醇提液中3种生物碱含量则有不同程度的降低,总生物碱也有不同程度的降低。与传统炮制品比较,醋烘法和微波炮制法中水提液的3种生物碱的含量均较高。结论延胡索经醋制和酒制后,水提液中有效成分的溶出率增大,以醋烘法和微波炮制法最佳。     

不同炮制方法对黄柏中小檗碱含量的影响

目的：探讨不同炮制方法对黄柏中小檗碱含量的影响,以得知黄柏的最佳炮制方法。以单波长锯齿扫描法测定了不同炮制方法之黄柏1%HCL甲醇提和水煎液中小檗碱的含量。结果：小檗碱的含量：盐炒甲醇提取0.531%;100℃烘后甲醇提取:0.597%;微波照射后甲醇提取：0.726%;盐炒后水煎提取:0.338%;100℃烘后水煎提取：0.447%;微波照射后水煎提取：0.476%。结论：黄柏中小檗碱的含量以微波法炮制为最高,故盐制黄柏应用微波法炮制。     

响应面法优化白屈菜醋焖法炮制工艺研究

摘要：目的:采用星点设计响应曲面法优选醋焖法炮制白屈菜的最佳炮制工艺。方法:建立原阿片碱、白屈菜碱、盐酸黄连碱、血根碱、小檗碱和白屈菜红碱等6种生物碱成分含量的HPLC法,色谱柱:Kromasil C18（250 mm×4.6 mm,5μm）;流动相:0.1%三乙胺（用磷酸调节pH至3.0）-乙腈,梯度洗脱;检测波长:274 nm;流速:1.0 ml·ml-1;柱温:35℃。以6种生物碱总含量为评价指标,采用响应曲面设计法,考察醋用量、闷润时间、烘干时间和烘干温度对白屈菜醋焖法炮制工艺的影响,确定最佳炮制工艺。结果:原阿片碱等6个对照品在0.640～12.800,1.024～20.480,0.862～17.240,0.436～8.720,0.226～4.520,0.324～6.480μg质量范围内线性关系良好（r值范围0.999 7～0.999 9）,平均加样回收率分别为97.5%,99.4%,100.8%,99.7%,98.9%和98.6%,RSD在1.82%～2.43%之间（n=6）。白屈菜总碱含量提取的最佳工艺条件为:醋用量17%,闷润时间2 h,烘干时间16h,烘干温度75℃。结论:所建立的白屈菜红碱等6种生物碱含量的HPLC法稳定可行。优选的醋焖法炮制白屈菜炮制工艺方法可行,为白屈菜的进一步研究提供了科学依据。     

白屈菜甘草制减毒炮制工艺研究

摘要：目的:建立甘草制白屈菜减毒增效炮制工艺。方法:以甘草用量、闷润时间、烘干时间和烘干温度等4个因素,利用正交试验进行炮制工艺研究,以别隐品碱等7个生物碱成分作为指标,采用HPLC法对炮制前后成分进行含量测定。结果:所建立的HPLC法符合相关规定。确定最佳工艺为A3B3C1D1,即甘草用量15%,闷润3 h, 60℃下烘12 h。结论:所建立的甘草制炮制工艺方法可行,提示本甘草制工艺具有一定的减毒作用。     

HPLC法同时测定川黄柏中盐酸黄柏碱和盐酸小檗碱的含量

目的:建立同时测定川黄柏中盐酸黄柏碱和盐酸小檗碱含量的方法。方法:采用高效液相色谱法。色谱柱为Waters XTERRA?MS C18(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸溶液(每100 ml中加0.1 g十二烷基磺酸钠,梯度洗脱),检测波长为284 nm(盐酸黄柏碱)和265 nm(盐酸小檗碱),流速为1.0 ml/min,柱温为30℃。结果:盐酸黄柏碱和盐酸小檗碱的质量浓度分别在9.45²36.15、74.77236.15、74.77¹ 869.25 mg/L范围内与各自峰面积积分值呈良好的线性关系(r分别为0.999 1、0.999 8);二者精密度、稳定性、重复性试验的RSD≤1.55%;平均加样回收率分别为97.47%1 869.25 mg/L范围内与各自峰面积积分值呈良好的线性关系(r分别为0.999 1、0.999 8);二者精密度、稳定性、重复性试验的RSD≤1.55%;平均加样回收率分别为97.47%⁹8.24%、95.03%98.24%、95.03%⁹5.63%,RSD分别为0.39%95.63%,RSD分别为0.39%⁰.93%、0.42%0.93%、0.42%⁰.93%(n均为3)。结论:该方法简便、快速、专属性强,可有效评价川黄柏中盐酸黄柏碱和盐酸小檗碱的质量。     

Synthesis and evaluation of 2,6-piperidinedione derivatives as potentially novel compounds with analgesic and other CNS activities

New 2,6-piperidinediones 2_a–g and 4_a–d were prepared by initial condensation of aromatic aldehydes or cycloalkanones with cyanoacetamide to give α-cyanocinnamides l_a–g or cycloalkylidenes 3_a,b which underwent Michae1 addition with ethyl cyanoacetate or diethylmalonate. Compounds 4_a–d were alkylated by various alkyl halides to produce the N-alkylated 2,6-piperidinedione derivatives 5_a–m. Some new selected compounds 2_a–c,f, 4_a–d & 5_e,h,j were pharmacologically evaluated for potential anticonvulsant, sedative and analgesic activities. These compounds exhibited significant anticonvulsant and analgesic effects after a single I.P. administration 100 mg/kg b.wt. . On the other hand all the investigated compounds induced hypnotic activity and prolonged the phenobarbital sodium- induced sleep as compared with the control group and the most potent compound was found to be 2_f.     

去壁灵芝孢子粉片的含量测定与免疫调节及心脏保护功能研究

目的 研究并评价“仙芝3号”去壁灵芝孢子粉片的免疫调节和心脏保护作用。方法 首先运用UPLC-Q-TOFMS和比色法分析“仙芝3号”去壁灵芝孢子粉片的主要化学成分,考察其对斑马鱼巨噬细胞减少症模型、斑马鱼心衰模型、H2O2诱导的心肌及内皮细胞氧化应激模型、脂多糖诱导的小胶质细胞炎症模型的影响,通过巨噬细胞形成能力、巨噬细胞吞噬能力、抗神经炎症能力、心脏收缩舒张改善能力、抗心肌及内皮细胞氧化应激损伤能力等多个指标评估其免疫调节及心脏保护作用。结果 “仙芝3号”去壁灵芝孢子粉片可以提高巨噬细胞形成及吞噬能力,改善心脏收缩舒张功能,减少神经炎症并缓解心肌及内皮细胞因氧化应激而造成的损伤,从而起到免疫调节和心脏保护作用。结论 “仙芝3号”去壁灵芝孢子粉片有望用于免疫功能调节及心脏功能保护。     

Tricalysiolide G and tricalysiols A and B: rearranged ent-kaurane-type and ent-kaurane-type diterpenoids from the leaves of Tricalysia dubia (Lindl.) Ohwi

Three rearranged ent-kaurane diterpenes with the cafestol-type framework have been isolated from the leaves of Tricalysia dubia. Two were found to be known diterpenoids, tricalysiolides B and C. Tricalysiolide B was isolated as colorless prisms in this experiment and its three-dimensional structure was determined by X-ray crystallography. The remaining diterpenoid was a new compound and was named tricalysiolide G. Two new ent-kaurane-type diterpenoids, given the trivial names tricalysiol A and tricalysiol B, were also isolated. The structures of the new compounds were elucidated from spectroscopic evidence.     

Metabolism and hepatotoxicity of N,N-dimethylformamide,N-hydroxymethyl-N-methylformamide,and N-methylformamide in the rat

The metabolism and hepatotoxicity ofN,N-dimethylformamide (DMF) and two of its metabolites,N-hydroxymethyl-N-methylformamide (HMMF) andN-methylformamide (NMF) were evaluated over a 4-day period in rats. DMF toxicity was dose dependent and delayed toxicity after the administration of a high DMF dose (13.7 mmol/kg) in comparison to a lower dose (4.1 mmol/kg) was observed. Treatment of rats with 13.7 mmol/kg DMF, HMMF, or NMF showed i) that DMF is more toxic than HMMF or NMR, and ii) that hepatotoxicity occurs later for DMF than for HMMF or NMF. Analysis of serum and urine samples demonstrated that DMF is first metabolized to HMMF, which is then partially converted to NMF. After HMMF administration, NMF was found both in serum and in urine. The time course of DMF and HMMF toxicity in relation to NMF formation fitted the hypothesis that the hepatotoxicity of DMF and HMMF is mediated via NMF. The degree of hepatotoxicity after HMMF and NMF treatment is similar. However, the degree of DMF hepatotoxicity is much higher than in the case of NMF or HMMF. The role of NMF as an obligatory intermediate in DMF and HMMF hepatotoxicity is discussed.     

Terbinafine susceptibility patterns for onychomycosis-causative dermatophytes and Scopulariopsis brevicaulis

The in vitro antifungal activity of terbinafine against 521 clinical isolates of seven species of dermatophytes, including four onychomycosis-causative species, as well as five Scopulariopsis brevicaulis isolates was determined by a modified Clinical and Laboratory Standards Institute microdilution method. Results showed a high antifungal activity of terbinafine against all dermatophyte isolates (geometric minimal inhibitory concentration (MIC)=0.026 microg/mL; concentration inhibiting 50% of mycological growth (MIC50)=0.03 microg/mL; and concentration inhibiting 90% of mycological growth (MIC90)=0.06 microg/mL). The geometric mean MICs against onychomycosis-causative dermatophyte species was lower (0.024 microg/mL) than the global MIC. However, the in vitro activity of terbinafine against S. brevicaulis was considerably lower (geometric mean MIC=1.38 microg/mL) in comparison with dermatophytes. The antifungal activity of itraconazole was lower than that of terbinafine against these fungi. These data confirm the high in vitro antifungal activity of terbinafine against dermatophytes, under standardised conditions.     

Investigation of the prevalence of tetQ, tetX and tetX1 genes in Bacteroides strains with elevated tigecycline minimum inhibitory concentrations

In this study, the antibiotic susceptibilities to tigecycline and tetracycline of 35 selected Bacteroides fragilis group strains were determined by Etest, and the presence of tetQ, tetX, tetX1 and ermF genes was investigated by polymerase chain reaction (PCR). tetQ was detected in all 12 B. fragilis group isolates (100%) exhibiting elevated tigecycline minimum inhibitory concentrations (MICs) (≥8 μg/mL) as well as the 8 strains (100%) with a tigecycline MIC of 4 μg/mL, whilst tetX and tetX1 were present in 15% and 75% of these strains, respectively. All of these strains were fully resistant to tetracycline (MIC ≥ 16 μg/mL). On the other hand, amongst the group of strains with tigecycline MICs < 4 μg/mL (15 isolates), tetQ, tetX and tetX1 were found less frequently (73.3%, 13.3% and 46.7%, respectively). All but two strains harbouring the tetQ gene in this group were non-susceptible to tetracycline, with a MIC > 4 μg/mL. These data suggest that in most cases tigecycline overcomes the tetracycline resistance mechanisms frequently observed in Bacteroides strains. However, the presence of tetX and tetX1 genes in some of the strains exhibiting elevated MICs for tigecycline draws attention to the possible development and spread of resistance to this antibiotic agent amongst Bacteroides strains. The common occurrence of ermF, tetX, tetX1 and tetQ genes together predicted the presence of the CTnDOT-like Bacteroides conjugative transposon in this collection of Bacteroides strains.     

Bioaccessibility of Hg, Cd and As in cooked black scabbard fish and edible crab

Regular consumption of seafood has been widely recommended by authorities. Yet, some species accumulate high levels of contaminants like Hg, Cd and As. In addition, the risks associated to the consumption of such seafood may increase if consumers use cooking practices that enhance the concentration of contaminants and their bioaccessibility. In this study, the bioaccessibility of Hg, Cd and As was assessed with in vitro human digestion of raw and cooked black scabbard fish (Hg; steamed, fried and grilled) and edible crab (Cd and As; steamed and boiled) tissues. Additionally, the toxicological hazards associated with the consumption of these products were also discussed. Generally, Hg, Cd and As bioacessibility increased throughout the digestion process. Cadmium and As revealed high bioaccessibility rates in raw and cooked samples (up to 100%), whereas lower bioaccessible fractions of Hg was observed (up to 40%). Furthermore, this study pointed out the importance of food matrix, elemental chemical properties and cooking practices in the bioaccessibility of Hg, Cd and As. The toxicological hazards revealed that edible crab brown meat (Cd) and grilled black scabbard fish (MeHg) consumption in children should be moderated. In contrast, edible crab muscle (Cd) and fried or steamed black scabbard fish (MeHg) should be consumed to minimize exposure. The use of bioaccessible contaminant data strongly reduced the toxicological risks of MeHg, whereas less risk reduction occurred with Cd and inorganic As.     

Metabolic activation of aromatic and heterocyclicN-hydroxyarylamines by wild-type and mutant recombinant human NAT1 and NAT2 acetyltransferases

Recombinant human NAT1 and polymorphic NAT2 wild-type and mutantN-acetyltransferases (encoded byNAT2 alleles with mutations at 282/857, 191, 282/590, 341/803, 341/481/803, and 341/481) were expressed inEscherichia coli strains XA90 and/or JM105, and tested for their capacity to catalyze the metabolic activation (viaO-acetylation) of theN-hydroxy (N-OH) derivatives of 2-aminofluorene (AF), and the heterocyclic arylamine mutagens 2-amino-3-methylimidazo [4,5-f]quinoline (IQ), 2-amino-3,4-dimethyl-imidazo[4,5-f]quinoxaline (MeIQx), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Both NAT1 and NAT2 (including all mutant human NAT2s tested) catalyzed the metabolic activation of each of theN-hydroxyarylamines to products that bound to DNA. Metabolic activation of N-OH-AF was greater than that of the heterocyclicN-hydroxyarylamines. The relative capacity of NAT1 versus NAT2 to catalyze activation varied withN-hydroxyarylamine substrate. N-OH-MeIQx and N-OH-PhIP exhibited a relative specificity for NAT2. These results provide mechanistic support for a role of the genetic acetylation polymorphism in the metabolic activation of heterocyclic amine mutagens and carcinogens.     

Photodegradation and phototoxicity of thioridazine and chlorpromazine evaluated with chemical analysis and aquatic organisms

The photochemical behaviour of chlorpromazine (CPZ) and thioridazine (THR) incubated under VIS light and a UV-A lamp was investigated with a high-performance liquid chromatography photodiode array detector (HPLC-PAD) and two bioassays. VIS light caused the decrease of CPZ and THR to 25% and 34% of the initial level, respectively, while UV-A degraded the drugs almost totally. CPZ and THR were very toxic to the protozoan Spirostomum ambiguum (Spirotox) and anostracan crustacean Thamnocephalus platyurus (Thamnotoxkit F^TM) with 24-h LC50 values of around 0.5 mg l⁻¹. In spite of the drastic decrease of the concentration of the drugs, the irradiated samples were toxic to the protozoan, especially when a sublethal end-point was taken into consideration. Contrary to the protozoan the crustacean was not sensitive to the products of photodegradation. Mass spectrometry analysis showed the presence of dimers and trimers of the CPZ and mono-, di-, and tri-oxygenated derivatives of THR. The presented data give a strong indication of the importance of the investigation of the environmental fate of drugs, especially those known to be phototoxic.     

Prevalence and characterization of the mechanisms of macrolide, lincosamide and streptogramin resistance in viridans group streptococci

The presence of erm genes conferring constitutive and inducible resistance, as well as that of the mefA gene conferring only constitutive resistance, was investigated using PCR in 70 erythromycin resistant (MIC≥1 mg/l) strains of viridans group streptococci (VGS) (18 Streptococcus mitis biotype 1, 16 S. mitis biotype 2, 15 S. oralis, 12 S. salivarius and nine S. sanguis) isolated from the oropharynx of healthy Greek children. All of the 56 isolates belonging to resistance phenotype M harbored the mefA gene. All of the 14 isolates constitutively resistant to macrolides and lincosamides (phenotype CR) harbored the ermB gene. Co-presence of both genes was not observed, whereas class A erm gene (previously known as ermTR) was not detected. Our results are consistent with a possible role of VGS as a reservoir of resistance genes now prevalent in pathogenic species of streptococci.