Chinese Medicine Qingluo Side Component Analysis

In clinical practice, traditional Chinese medicine (TCM) is characterized by Chinese herbal formula usually consisting of more than two herbs, which reflects the TCM rule of treating diseases by the differentiation of ZHENG ("Bian Zheng Lun Zhi") as well. Thus, research work of herbal formula contributes a lot to the development of TCM. In the present study, an anti-angiogenesis herbal formula, QingLuo Fang (QLF), is employed to address this key point. Modern analytical technologies of reversed-phase high-performance liquid chromatography (HPLC) and LC-MSn are used to analyze the components of QLF in vitro and in vivo.Firstly, we developed a method using HPLC-ultraviolet detector for simultaneous determination of five bioactive alkaloids (matrine, oxymatrine, oxysophocarpine, sinomenine and berberine) in the QLF decoction. We carefully selected the HPLC column, modulated components and the pH values of the mobile phase, and adjusted the gradient procedure. Then, an efficient and quick analysis within 28 minutes was achieved for the five bioactive alkaloids in QLF. Next, the influences on the chromatographic behavior of each alkaloid caused by different pH values were explored, which may benefit the related chromatographic theory. Moreover, the developed method was also applied to analyze the five alkaloids in decoction of individual herbs, and comparisons were made to evaluate the inter-herb influences in the formula.Secondly, to clarify the compounds derived from QLF that absorbed in rat blood, a HPLC-MSn method was employed to systemically analyze the QLF decoction, the decoction of each component herb, and rat blood after the administration of QLF. In this work, we compared the HPLC-MSn information about QLF and each component herb to classify each of the detected compounds to the corresponding herbs at first. Next, we obtained the association of QLF with the detected compounds by revisited literatures about each herb in QLF. Then, we gained the new compounds presenting in rat blood after the administration of QLF at 20min, 1h and 2h, respectively. Lastly, by the comparison of in vitro and in vivo, we defined the following m/z information about compounds derived from QLF and absorbed in rat blood: 205 (N-methylcytisine), 265 (Sophoranol or some compound with a molecular weight of 264 from Root Sophora), 249 (1) (Sophoridine or Iosmatrine or lupanine), 330 (Sinomenine), 249 (2) (Matrine) and 247 (Sophocarpine). Interestingly, when administrated after 20min, 1h and 2h, two peaks with the mass number of 293 and 316 are observed which were undetectable in QLF. These two peaks are speculated to be the transforms in vivo of Berberine, other components of QLF, or even the endogenous factors of rat blood stimulated by QLF. The peak of 293 is likely the in vivo transforms of Berberine since one peak at m/z = 292 falls into 2 to 3 MS cataclastic...