Development and Validation of an HPLC Method for Karanjin in Pongamia pinnata linn. Leaves

A rapid, simple and specific reversed-phase HPLC method has been developed for analysis of karanjin in Pongamia pinnata Linn. leaves. HPLC analysis was performed on a C₁₈ column using an 85:13.5:1.5 (v/v) mixtures of methanol, water and acetic acid as isocratic mobile phase at a flow rate of 1 ml/min. UV detection was at 300 nm. The method was validated for accuracy, precision, linearity, specificity. Validation revealed the method is specific, accurate, precise, reliable and reproducible. Good linear correlation coefficients (r²>0.997) were obtained for calibration plots in the ranges tested. Limit of detection was 4.35 μg and limit of quantification was 16.56 μg. Intra and inter-day RSD of retention times and peak areas was less than 1.24% and recovery was between 95.05 and 101.05%. The established HPLC method is appropriate enabling efficient quantitative analysis of karanjin in Pongamia pinnata leaves.     

Red cell haemolysis test as an in vitro approach for the assessment of toxicity of karanja oil

The karanja tree grows in parts of India and Australia. The oil from seed kernels was found to be toxic to animals. The annual potential availability of the oil is around 135,000 tons in India. In order to use it for beneficial purposes, it is necessary to detoxify the oil. In the present study, the oil was assessed for toxicity by the red cell haemolysis test and estimating the LDH in the supernatant. The non-lipid constituents were isolated from raw oil by aqueous methanol extraction. The raw oil and the non-lipid fraction were found to haemolyse the red cells with release of LDH, whereas the extracted oil did not show such a manifestation. There was a good correlation between haemolytic activity and LDH released from cells. These findings were further confirmed with in vivo studies where the raw and extracted karanja oils showed 100% and nil mortality in rats dosed orally at 10 and 20 ml/kg body weight, respectively. This haemolysis test can be used as an in vitro method to predict toxicity and to monitor the detoxification of the oils prior to use in in vivo studies for toxicological evaluation. The fatty acid composition of the raw and extracted karanja oils showed no difference.     

Red cell haemolysis test as an in vitro approach for the assessment of toxicity of karanja oil.

The karanja tree grows in parts of India and Australia. The oil from seed kernels was found to be toxic to animals. The annual potential availability of the oil is around 135,000 tons in India. In order to use it for beneficial purposes, it is necessary to detoxify the oil. In the present study, the oil was assessed for toxicity by the red cell haemolysis test and estimating the LDH in the supernatant. The non-lipid constituents were isolated from raw oil by aqueous methanol extraction. The raw oil and the non-lipid fraction were found to haemolyse the red cells with release of LDH, whereas the extracted oil did not show such a manifestation. There was a good correlation between haemolytic activity and LDH released from cells. These findings were further confirmed with in vivo studies where the raw and extracted karanja oils showed 100% and nil mortality in rats dosed orally at 10 and 20 ml/kg body weight, respectively. This haemolysis test can be used as an in vitro method to predict toxicity and to monitor the detoxification of the oils prior to use in in vivo studies for toxicological evaluation. The fatty acid composition of the raw and extracted karanja oils showed no difference.     

薄层色谱-荧光分析法测定水罗伞药材中水黄皮素含量的研究

目的建立薄层色谱-荧光分光光度法测定水罗伞中水黄皮素含量的方法。方法采用无水乙醇为溶剂,硅胶G为薄层板,苯-乙酸乙酯(8.5∶1.5)为展开剂,激发波长与荧光波长分别为355nm和428nm。结果水黄皮素线性范围为0.138～0.69μg,r=0.9991,平均回收率为99.14%,RSD为1.94%(n=5)。结论该法便捷、灵敏、准确、重现性好,可以控制该药材的质量。     

基于网络药理学和分子对接的薏苡附子散对类风湿性关节炎及慢性心力衰竭“异病同治”作用机制探讨

目的 基于网络药理学和分子对接方法探讨薏苡附子散治疗类风湿性关节炎(RA)及慢性心力衰竭(CHF)"异病同治"的作用机制。方法 通过中药系统药理学数据库与分析平台(TCMSP)数据库筛选薏苡附子散的活性成分,并利用 Pubchem 数据库获得 Smile 号,进而通过 Swiss Target Prediction 数据库获取活性成分对应的靶点。在 GeneCards 和 OMIM数据库平台预测 RA 及 CHF 疾病作用靶点。对活性成分靶点与疾病交集靶点进行蛋白质相互作用(PPI)网络构建及核心靶点筛选,并进行基因本体(GO)功能及京都基因与基因组百科全书(KEGG)通路富集分析。筛选核心成分和核心靶点,利用 AutoDock 软件进行分子对接验证。结果 从薏苡附子散中共筛选得到符合条件的活性成分 10 个,活性成分潜在作用靶点 403 个,RA 疾病靶点 1 112 个,CHF 疾病靶点 2 983 个,取交集,获得药物、疾病交集靶点 69 个。GO 富集分析结果显示主要与炎症反应的调节、MAPK 级联调控、对氧化应激的反应、骨化的调节、平滑肌细胞增殖等相关;KEGG 通路富集分析得到 236 条通路,与RA及CHF相关且排序较靠前的通路有 PI3K-Akt 信号通路、NF-κB 信号通路、MAPK 信号通路等。分子对接结果表明,薏苡附子散中的核心成分多根乌头碱、水黄皮素、豆甾醇具有与核心靶点 TNF、SRC、ESR1 和 PTGS2良好的结合潜力。结论 薏苡附子散治疗 RA 和 CHF 具有多靶点效应,涉及多条生物过程和信号通路,可能通过调控炎症反应、氧化应激反应、血管生成和发育、心脏收缩调节及 MAPK 级联调控等发挥"异病同治"作用。