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首页> 外文期刊>American Journal of Analytical Chemistry >Quantitation of Polydeoxyribonucletides (PDRNs) in Human Placental Extract by Fluorescence Spectroscopy Using Ethidium Bromide
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Quantitation of Polydeoxyribonucletides (PDRNs) in Human Placental Extract by Fluorescence Spectroscopy Using Ethidium Bromide

机译：溴化乙锭荧光光谱法测定人胎盘提取物中的聚脱氧核糖核酸（PDRNs）

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摘要

Characterization of an aqueous extract of human placenta, used as a licensed drug for wound healing, leads to the identification of several bioactive components including polydeoxyribonu-cleotides (PDRNs). PDRNs are mixture of DNA fragments of different molecular weight. A spectro-fluorimetric method of quantitation of PDRNs in the aqueous extract of human placenta by using ethidium bromide (EtBr) has been described here. It has been demonstrated by thin layer chromatography (TLC) followed by reversed phase HPLC that EtBr binds specifically with the PDRN fraction of the multi-component extract. The binding specificity of EtBr has been verified by the analysis of emission spectra of the extract. A concentration of 0.29 μg/ml EtBr exhibits a linear range of standard CT-DNA from 0.5 - 5 μg/ml of buffer (R2 = 0.992). The same concentration of EtBr shows a linear range of measurements of placenta extract from 5 - 35 μl/ml of buffer (R2 = 0.976). The points of the curve were the average of three sets where maximum variation observed was ±3%. PDRN content of the extract has been estimated based on the resultant fluorescence emission (after background correction) with respect to the standard calibration curve of calf thymus DNA (CT-DNA). Estimation of PDRN in a large number of batches of placenta extract (n = 100) has been done. The statistical analysis of the estimation was found to be significant and the lower and upper levels of PDRN were 158.30 and 239.03 μg/ml of the extract respectively. This easy-to-use method of estimation of PDRN in multi-component biological extract is reported for the first time. This will help in quantitation of PDRNs for other biological extracts.
机译：用作伤口愈合许可药物的人胎盘水提取物的表征导致鉴定了几种生物活性成分，包括聚脱氧核糖核苷酸（PDRN）。 PDRN是不同分子量的DNA片段的混合物。本文介绍了使用溴化乙锭（EtBr）的荧光定量荧光法定量人胎盘水提物中的PDRN。通过薄层色谱法（TLC），然后通过反相HPLC证明，EtBr与多组分提取物的PDRN组分特异性结合。 EtBr的结合特异性已通过分析提取物的发射光谱进行了验证。浓度为0.29μg/ ml的EtBr具有0.5-5μg/ ml缓冲液的线性CT-DNA线性范围（R2 = 0.992）。相同浓度的EtBr显示5-35μl/ ml缓冲液中胎盘提取物的线性测量范围（R2 = 0.976）。曲线的点是三组的平均值，观察到的最大变化为±3％。提取物的PDRN含量是根据相对于小牛胸腺DNA（CT-DNA）的标准校准曲线的荧光发射（背景校正后）估算的。大量胎盘提取物（n = 100）中PDRN的估算已经完成。估计的统计分析被认为是有意义的，PDRN的较低和较高水平分别为提取物的158.30和239.03μg/ ml。首次报道了这种易于使用的估算多组分生物提取物中PDRN的方法。这将有助于定量其他生物提取物的PDRN。

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