基于电喷雾质谱法及化学计量法筛选鹿茸促睾丸间质细胞分泌睾酮的活性成分

采用电喷雾串联质谱法(ESI-MS/MS)及化学计量法快速筛选鹿茸促睾丸间质细胞分泌睾酮的活性成分。运用电喷雾串联质谱法分别检测6批鹿茸鲜品和6批鹿茸传统加工品中化学成分,并测定各样品增加大鼠睾丸间质细胞的睾酮分泌量,联合主成分分析(PCA)、偏最小二乘法判别分析(PLS-DA)和正交偏最小二乘法判别分析(OPLS-DA)对得出的一级质谱数据进行分析;实现鹿茸鲜品和鹿茸传统加工品两组样品的快速区分。采用偏最小二乘法(PLS)和灰色关联度分析(GRA)筛选出其中具有潜在促睾丸间质细胞分泌睾酮的活性成分,通过对二级质谱数据分析明确其中的可能活性成分的结构和组成。结果表明,鹿茸鲜品与加工品的组成成分有显著差异,且二者均具有良好的促睾丸间质细胞分泌睾酮的活性,其中活性较强的成分主要为Tyr-Ser-Phe(m/z416)和Phe-Phe-Leu/Ile(m/z 426)。本方法能够简单快速分析鹿茸中促睾丸间质细胞的活性成分,为鹿茸作为保健食品的后续开发提供参考。

The Active Components of Testosterone Secreted by Antler Testis Stromal Cells Rapidly Screened by Electrospray Tandem Mass Spectrometry and Chemometrics

Electrospray tandem mass spectrometry (ESI-MS/MS) and chemometrics were used to screen the active components of testosterone secreted by testis stromal cells of Antler antler. Using electrospray tandem mass spectrometry, the chemical composition of 6 batch of pilose antler and 6 batch of velvet antler prepared by traditional process was detected. The testosterone secretion of rat leydig cells was also determinated. The principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to analyze the mass spectrometry data. Rapid differentiation of fresh velvet antler and traditional processed products was achieved. Partial least square method (PLS) and gray correlation degree analysis (GRA) were used to select the active components with potential testosterone secretion by testicular stromal cells. The structure and composition of the possible active components were determined by secondary mass spectrometry data analysis. The results showed that the components of fresh antler and traditional processed products were significantly different, and both of them had good activity of testosterone secretion by testicular stromal cells. The most active components were Tyr-Ser-Phe (m/z 416), PHE-Phe-Leu/Ile (m/z 426), respectively. This method could be used to analyze the active components of testicular stromal cells in antler and provide reference for the development of antler as health food.