HPLC柱切换法测定抗癌药米托蒽醌血浆浓度

新抗癌药米托蒽醌治疗剂量低,与血浆蛋白结合率较高,用一般样品预处理技术,操作麻烦费时。本文采用HPLC柱切换技术,以甲醇沉淀血浆蛋白,经超声振荡10 min,以YWG-CN为预处理柱(5cm×4.6 mm ID),水为预处理流动相,在线富集样品;以Shimpack CLC-ODS为分析柱(15cm×6 mm ID),甲醇—乙酸铵缓冲液(0.2 mol/L,pH 1.9)(48:52)为流动相,658 nm检测,内标法定量。净化富集样品操作简便,回收率好(85.5%);米托蒽醌的检测限为0.5 ng。血浆中最低检出浓度为6 ng/ml。日内变异系数为2.8～11%,日间变异系数为4.0～13.8%。在浓度为10～1000 ng/ml血浆范围内,呈线性关系。     

紫杉烷二萜类化合物10-DABⅢ的分离纯化

目的:建立分离纯化紫杉烷二萜类化合物10-脱乙酰基巴卡亭Ⅲ(10-DABⅢ)的方法。方法:采用柱层析和重结晶的方法精制10-DABⅢ。以杂质含量为指标确定层析液比例;以纯度和收率为指标筛选第1次重结晶时三氯甲烷的用量,第2次重结晶时丙酮、正己烷的用量以及2次重结晶的温度和时间。结果:层析液丙酮与三氯甲烷体积比为2.25:7.75,此时杂质含量最小,为3.7%。用15倍体积(W/V)的三氯甲烷、-25℃冷冻24h第1次重结晶以及35倍体积的丙酮(W/V)和30倍体积的正己烷(W/V)、-25℃冷冻12h第2次重结晶,纯化后的样品纯度为99.34%,收率为83.50%。结论:该方法对10-DABⅢ分离纯化效果好。     

毛细管气相色谱法测定水中2,4,6-三氯酚和五氯酚

目的:建立测定水中2,4,6-三氯酚和五氯酚的气相色谱测定方法。方法:水中的微量2,4,6-三氯酚和五氯酚经液-液萃取后,用DB-1701毛细管色谱柱,在程序升温条件下进行GC分析,以保留时间定性,外标法定量。结果:2,4,6-三氯酚的线性范围为0.005μg/L~1.0μg/L,最低检出浓度为0.0030μg/L,五氯酚的线性范围为0.01μg/L~2.0μg/L,最低检出浓度为0.0048μg/L,相关系数均≥0.9990,水样加标回收率为73.0%~91.0%,RSD为4.6%~7.9%。结论:方法仅用8.5 m in完成饮用水中2,4,6-三氯酚和五氯酚的测定,操作简便、灵敏度高。     

幽门螺杆菌尿素酶抗原的分离和纯化

本研究采用层析技术，以ＳｅｐｈａｄｅｘＧ２００分离幽门螺杆菌尿素酶，从幽门螺杆菌超声破碎物离心上清中一次提纯了尿素酶抗原，产物接近电泳纯并保持了良好的抗原性。我们对５５份抗幽门螺杆菌抗体阴性村本和５０分阳性标本检测发现，符合率达１００％，假阴性和假阳性均为零；大样本临床标本检测也取得了很好的结果。     

藏边大黄的化学成分研究

目的 研究蓼科波叶组藏边大黄的化学成分.方法 采用硅胶、聚酰胺和ODS柱层析法.结果 分离得到11个化合物,经化学方法和波谱学鉴定,确定结构分别为:大黄酚(Ⅰ)、大黄素甲醚(Ⅱ)、大黄素(Ⅲ)、β-谷甾醇(Ⅳ)、trans-3,5,3′,4′-四羟基芪(Ⅴ)、d-儿茶素(Ⅵ)、胡萝卜苷(Ⅶ)、trans-3,5,3′,4′-四羟基芪-4′-O-β-D-吡喃葡萄糖苷(Ⅷ)、trans-3,5,3′,4′-四羟基芪-4′-O-β-D-(6″-O-没食子酰)-吡喃葡萄糖苷(Ⅸ)、大黄素-8-O-β-D-吡喃葡萄糖苷(X)和大黄酚-8-O-β-D-吡喃葡萄糖苷(Ⅺ).结论 其中Ⅸ系首次从该植物中分得.     

Oxidation Potentials of Flavonoids Determined by Flow-through Column Electrolysis

1　ＩｎｔｒｏｄｕｃｔｉｏｎＦｌａｖｏｎｏｉｄｓａｒｅｎａｔｕｒａｌｃｏｍｐｏｕｎｄｓ ,ｗｉｄｅｌｙｄｉｓｔｒｉｂｕｔｅｄｉｎｆｒｕｉｔｓ,ｖｅｇｅｔａｂｌｅｓ,ａｎｄｔｅａｓ.Ｔｈｅｒｅｉｓａｎｉｎｃｒｅａｓｉｎｇｉｎｔｅｒｅｓｔｉｎｔｈｅｓｅｃｏｍｐｏｕｎｄｓｂｅｃａｕｓｅｏｆｔｈｅｉｒｂｒｏａｄ ｐｈａｒｍａｃｏｌｏｇｉｃａｌａｃｔｉｖｉ ｔｉｅｓ ,[1,2 ] ｅｓｐｅｃｉａｌｌｙｔｈｅｉｒａｎｔｉｏｘｉｄａｎｔａｃｔｉｖｉｔｙ .[3～ 5 ] Ａｓａｎｔｉｏｘｉｄａｎｔｓ ,ｆｌａｖｏｎｏｉｄｓｈａｖｅｔｈｅａｂｉｌｉｔｙｔｏｓｃａｖｅｎｇｅｆｒｅｅｒａｄｉｃａｌｓｂｙｐｒｏｔｅｉｎａｎｄｅｌｅｃｔｒｏｎｔｒａｎｓｆｅｒｐｒｏｃｅｓｓｅｓ.Ｏｎｔｈｅｏｔｈｅｒｈａｎｄ ,ａｓｍｅｔａｌ ｃｈｅｌａｔｉｎｇａｇｅｎｔｓ ,ｆｌａｖｏｎｏｉｄｓｃａｎｄｅｐｒｅｓｓｔｈｅｓｕｐｅｒｏｘｉｄｅ ｄｒｉｖｅｎＦｅｎｔｏｎｒｅａｃｔｉｏｎ .[6 ] Ｔｈｅｒｅａｒｅａｌｓｏｒｅｐｏｒｔｓｏｆｆｌａｖｏｎｏｉｄｓｉｎｈｉｂｉｔｉｎｇｔｈｅａｃｔｉｖｉｔｉｅｓｏｆａｎａｒｒａｙｏｆｅｎｚｙｍｅｓ.[1,7,...     

离子交换法分离纯化酶反应液中5-氟尿苷的研究

为开发从酶反应液中分离纯化5-氟尿苷的方法,研究了利用阴离子交换树脂以及硅胶柱色谱实现转化产物分离纯化的工艺;实验采用HPLC为检测方法,以5-氟尿苷含量为考察指标,从吸附量,洗脱率,回收率的角度考察了各种条件的影响。最终确定以D301树脂为吸附剂;最佳吸附条件为动态吸附,温度25℃,pH 8;动态洗脱,洗脱剂5%NaCl溶液,体积流量为3BV/h;再利用硅胶柱色谱,采用200～300目硅胶,以正己烷∶乙酸乙酯=12∶88作为洗脱剂,洗脱后结晶可得到5-氟尿苷纯品,色谱纯,最终收率72.4%。     

A validated stability-indicating LC method for estimation of etoposide in bulk and optimized self-nano emulsifying formulation: Kinetics and stability effects

The present investigation was aimed to establish a validated stability-indicating liquid chromatographic method for the estimation of etoposide (ETP) in bulk drug and self-nano emulsifying formulation. ETP was successfully separated from the degradation products formed under stress conditions on LiChrospher 100 C₁₈ reverse-phase column (a 250 mm × 4.6 mm i.d., 5-μm particle size) using 55:45 (v/v) acetonitrile–phosphate buffer saline (pH 4.5) as the mobile phase, at a flow rate of 1.0 mL min⁻¹ and detection at 283 nm. The response was a linear function of analyte concentration (R² > 0.9997) over the concentration range of 0.05–50 μg mL⁻¹. The method was validated for precision, accuracy, robustness, sensitivity and specificity. The % recovery of ETP at three different levels (50%, 100% and 150%) ranged between 93.84% and 100.06% in optimized self-nano emulsifying formulation, Etosid® soft-gelatin capsule and Fytosid® injection. First-order degradation kinetics of ETP were observed under acidic and alkaline conditions. The method was also applied for the stability assessment of self-nano emulsifying formulation under accelerated conditions, the formulation was found to be stable at all storage conditions with the shelf-life of 2.37 years at 25 °C. The method holds promise for routine quality control of ETP in bulk, pharmaceutical formulations as well as in stability-indicating studies.     

Biological activities of Pseudevernia furfuracea (L.) Zopf extracts and isolation of the active compounds

Ethnopharmacological relevance

Pseudevernia furfuracea (L.) Zopf (Parmeliaceae) is a common epiphytic lichen in the conifer-hardwood forest of Anatolia. This species is used in traditional medicine in Turkey as a treatment for wounds, eczema and hemorrhoids.

Aim of the study

The present study was designed to investigate the active compounds from Pseudevernia. furfuracea, and the isolation studies yielded atraric acid ( Aslan et al., 2006) as the major compound and a mixture of methyl hematommate ( Baumann, 1960) and methyl chlorohematommate ( Bay?r et al., 2006). Furthermore, methanolic extract from thalli of Pseudevernia. furfuracea and its fractions and isolates ( 1, 2 and 3) were investigated for in vitro antimicrobial and antioxidant activities, and in vivo antinociceptive, anti-inflammatory and wound healing activities.

Material and methods

Antimicrobial activities of the samples were determined by using the disc diffusion technique. 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was used as a rapid TLC screening method to evaluate the antioxidant activity of Pseudevernia. furfuracea. The thiobarbituric acid (TBA) test was used to assess the efficacy of the extracts in protecting liposomes from lipid peroxidation. In vivo inhibitory effect of the extracts on the carrageenan-induced hind paw edema model in mice was studied for the assessment of anti-inflammatory activity. p-Benzoquinone-induced abdominal constriction test was used to explore the antinociceptive effects of the extracts. Moreover, the wound healing potential of the plant extracts that were evaluated by using in vivo incision and excision wound models on rats and mice, were comparatively assessed with a reference ointment Madecassol^®.

Results

Significant antimicrobial activities were observed against Gram (+) microorganisms and Candida krusei and Candida. dubliniensis in dichloromethane (DCM) and ethyl acetate (EtOAc) extracts and isolates. The methanol (MeOH), DCM and EtOAc extracts of the lichen were found to possess moderate inhibitory activity on lipid peroxidation. Methanolic extract of the lichen was found to possess significant inhibitory activity on the carrageenan-induced hind paw edema model in mice whereas the other fractions did not show any activity. While DCM and EtOAc extracts and fractions showed notable anti-inflammatory activity on carrageenan-induced hind paw edema model without inducing any apparent acute toxicity or gastric damage. Moreover, topical application of the ointment prepared with MeOH extract and EtOAc fraction onto the incised wounds exerted remarkable wound healing activity.

Conclusion

The results of these experimental studies exhibited that nonpolar fractions of Pseudevernia. furfuracea have significant antimicrobial activity against especially Candida species and polar fractions (especially MeOH) display antioxidant, anti-inflammatory, antinociceptive and wound healing activities.     

Single versus double column fixation in transverse fractures of the acetabulum: A randomised controlled trial

IntroductionBiomechanical studies have compared fixation methods in transverse acetabular fractures, yet there is not enough clinical data to suggest an optimal fixation method. The aim of this randomized controlled trail was to compare fracture stability in posterior plating alone versus posterior plating and anterior column lag-screw fixation in treatment of transverse and transverse with posterior wall acetabular fractures.MethodsThirty patients were randomized to one of two groups, either posterior fixation alone (single column group), or posterior plating and anterior fixation with percutaneous anterior column screw (double column group). Patients were followed up with serial radiographic assessments documenting any loss of reduction, utilizing Matta’s radiological criteria, measuring the roof arc angles and by measuring any change in the femoral head offset.ResultsFifteen patients were randomized to each group. Mean patient age was 31 years, mean follow up period was 19 months (range 12–24). There was no significant differences between the two groups with regards the quality of post-operative reduction, blood loss, hospital stay and functional score using the modified Merle D'Aubinge and Postel score. The operative time was significantly longer in the double column fixation group (130 min versus 104 min). There was no loss of reduction observed in either of the two groups.ConclusionSingle poster column fixation in transverse and transverse posterior wall fractures showed similar result to double column fixation, in terms of fractures stability in the follow up period, quality of reduction and early functional outcome.