Study on Active Constituents of Rehmanniae Radix Praeparata Based on HPLC Multi Wavelength Fusion Fingerprint and Spectral Effect Relationship

OBJECTIVE To establish HPLC multi-wavelength fusion fingerprints of Rehmanniae Radix Praeparata and to screen the anti-diabetic osteoporosis active ingredients. METHODS The multi-wavelength(203, 210, 254, 334 nm) fusion fingerprints of 10 batches of Rehmanniae Radix Praeparata were established by HPLC. Through the efficacy evaluation of the diabetic osteoporosis rat model, the partial least squares regression method was used, the main active components were obtained and the efficacy was verified by the high glucose osteoblasts model, and the anti-diabetic osteoporosis effect of Rehmanniae Radix Praeparata was investigated. RESULTS A total of 26 common peaks were identified in the fusion fingerprints of 10 batches of Rehmanniae Radix Praeparata, and 17 peaks were positively correlated with the bone formation marker(serum alkaline phosphatase activity). Regression coefficients and VIP values of peak 8(catalpol) and peak 12(rehmannioside D) were the most significant. Both osteoblast proliferation activity and cellular alkaline phosphatase levels. CONCLUSION Catalpol and rehmannioside D have significant anti-diabetic osteoporotic effect, and might be the main active components of Rehmanniae Radix Praeparata. The multi-wavelength fusion fingerprint combined with the spectral effect relationship can be used as an effective strategy for the screening of active ingredients.