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.

Abstract

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 μg/mL for β-ecdysterone, 0.0035-130 μg/mL for 25S-inokosterone, 0.004-423 μg/mL for ginsenoside Ro, 0.0036-66 μg/mL for chikusetsusaponin IV and 0.0044-111 μg/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.