Stomatological Hospital, Xi'an Jiaotong University, Xi'an 710 004, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Sep 1;904:115-20. doi: 10.1016/j.jchromb.2012.07.031. Epub 2012 Aug 4.
We have developed an online analytical method that combines human periodontal ligament cell membrane chromatography (hPDLC/CMC) with high-performance liquid chromatography and mass spectrometry (LC/MS) for recognizing and identifying osteoplastic active components from Coptidis Rhizoma. Retention fractions on hPDLC/CMC were enriched onto an enrichment column and the components were directly analyzed by combining a 10-port column switcher with an LC/MS system for separation and preliminary identification. Using simvastatin (SIM) as a positive control, berberine from Coptidis Rhizoma was identified as the active component which could act on the hPDLC. The MTT colorimetric assay, alkaline phosphatase (ALP) activity, and staining tests revealed that berberine could promote hPDLC growth, increase the secretion of ALP in the culture medium, and enhance the formation of mineralized nodule, thus it is a potential osteoplastic ingredient. This hPDLC/CMC-online-LC/MS method can be applied for screening active components acting on hPDLC from traditional Chinese medicines exemplified by Coptidis Rhizoma and will be of great utility in drug discovery using natural medicinal herbs as a source of leading compounds.
我们开发了一种在线分析方法，将人牙周膜细胞膜色谱（hPDLC/CMC）与高效液相色谱和质谱（LC/MS）相结合，用于识别和鉴定黄连中的成骨活性成分。在 hPDLC/CMC 上的保留分数被富集到富集柱上，并用 10 通阀柱切换器与 LC/MS 系统相结合直接分析，用于分离和初步鉴定。以辛伐他汀（SIM）为阳性对照，鉴定黄连中的小檗碱为作用于 hPDLC 的活性成分。MTT 比色法、碱性磷酸酶（ALP）活性和染色试验表明，小檗碱可促进 hPDLC 生长，增加培养基中 ALP 的分泌，并增强矿化结节的形成，因此它是一种有潜力的成骨成分。这种 hPDLC/CMC-在线-LC/MS 方法可用于从黄连等中药中筛选作用于 hPDLC 的活性成分，对于以天然草药为先导化合物来源的药物发现将非常有用。
