Determination of major components from Radix Achyranthes bidentate using ultra high performance liquid chromatography with triple quadrupole tandem mass spectrometry and an evaluation of their anti-osteoporosis effect in vitro
Determination of major components from Radix Achyranthes bidentate using ultra high performance liquid chromatography with triple quadrupole tandem mass spectrometry and an evaluation of their anti-osteoporosis effect in vitro
复制标题
超高效液相色谱-三重四极杆串联质谱法测定牛膝中主要成分并评价其体外抗骨质疏松作用
DOI：
10.1002/jssc.201900146
复制
发表时间：
2019-07-01
影响因子：
3.1
通讯作者：
Cai, Baochang
中科院分区：
文献类型：
--
作者：
Tao, Yi;Huang, Surun;Cai, Baochang
蜕皮甾酮和皂苷是牛膝中最具特色的成分，作用于人体，促进胶原蛋白合成，刺激细胞生长。然而，这些成分与MC 3 T3-E1成骨细胞分化之间的关系尚不清楚。建立了超高效液相色谱-三重四极杆串联质谱法直接测定牛膝生品和盐炙品中1种蜕皮甾酮和4种皂苷的含量。对该方法的线性、日内和日间精密度、重复性、稳定性和回收率进行了研究。该方法在0.003-336 μ g/mL的β-蜕皮甾酮、0.0035-130 μ g/mL的25 S-肌醇甾酮、0.004-423 μ g/mL的异黄酮、0.0036-66 μ g/mL的竹皂苷IV和0.0044-111 μ g/mL的竹皂苷IVa的浓度范围内呈线性。日内和日间精密度均在2.7%以内。标准加入法测定各组分的回收率（98.7-102.5%）。该方法已成功地应用于10批牛膝生品和盐炙品中5种成分的同时测定。随后，进行这些提取物对MC 3 T3-E1成骨细胞的分化的检查。最后，采用典型相关分析方法，对五种成分的含量与其抗骨质疏松作用的关系进行了研究。
Ecdysterone and saponins are the most characteristic components of Radix Achyranthes bidentate, which acts on the human body to promote collagen synthesis and stimulates cell growth. However, the relationship between these components and the differentiation of MC3T3-E1 osteoblastic cells is unknown. We developed a rapid ultra high performance liquid chromatography with triple quadrupole tandem mass spectrometry method for direct determination of one ecdysterone and four saponins in crude and salt-processed Radix Achyranthes bidentate. The method was interrogated in terms of linearity, intra- and inter-day precision, repeatability, stability and recovery. The method was linear within the concentration ranges of 0.003-336 mu g/mL for beta-ecdysterone, 0.0035-130 mu g/mL for 25S-inokosterone, 0.004-423 mu g/mL for ginsenoside Ro, 0.0036-66 mu g/mL for chikusetsusaponin IV and 0.0044-111 mu g/mL for chikusetsusaponin IVa. The intra-and inter-day precisions were all within 2.7%. The standard addition method determined recovery rates for each component (98.7-102.5%). The method was successfully applied to simultaneously quantify five components in ten batches of crude and salt-processed Radix Achyranthes bidentate. Subsequently, the examination of these extracts on the differentiation of MC3T3-E1 osteoblastic cells were carried out. Finally, the relationships between the contents of five components and their anti-osteoporosis effect were investigated by using canonical correlation analysis.