Résumé
Objective:To establish the quality evaluation methods of Asparagi Radix decoction pieces and its standard decoction. Method:Ten batches of Asparagi Radix standard decoction were prepared. High performance liquid chromatography-evaporative light scattering detection method (HPLC-ELSD) was established for the determination of protodioscin and protoneodioscin in Asparagi Radix decoction pieces and its standard decoction, and the fingerprint detection of Asparagi Radix decoction pieces with acetonitrile-water as mobile phase for gradient elution. UHPLC-LTQ-Orbitrap-MS/MS was used to identify ten main common peaks in the fingerprint with acetonitrile-0.1% formic acid solution as mobile phase for gradient elution, electrospray ionization (ESI) and positive and negative ion mode scanning were employed, the detection range was m/z 100-1 400. Result:The total content of protodioscin and protoneodioscin in Asparagi Radix decoction pieces was 0.41%-0.72%, and their total content in Asparagi Radix standard decoction was 0.33%-0.59%, the transfer rate of these two components was 73.6%-98.3%. The dry extract yield of the standard decoction was 59.0%-73.0%, and its pH was 4.9-5.6. There were 10 common peaks in the fingerprint, and all of them were saponins, including protoneodioscin, protodioscin, aspacochioside A and its isomer, methyl protodioscin, asparagoside F, (25R)-26-O-β-D-glucopyranosyl-furostan-5, 20-diene-3β, 26-diol-3-O-[α-L-rhamnopyranosyl (1→2)]-[β-D-glucopyranosyl (1→4)-α-L-rhamnopyranosyl (1→4)]-β-D-glucopyranoside, 26-O-β-D-glucopyranosyl-furostan-20 (22)-ene-3β, 26-diol-3-O-[α-L-rhamnopyranosyl (1→2)]-[α-L-rhamnopyranosyl (1→4)]-β-D-glucopyranoside, pseudodiosgenin, aspacochioside C. Conclusion:In this paper, the quality evaluation methods of Asparagi Radix decoction pieces and its standard decoction are established, and these methods are stable and feasible, which can provide reference for the quality control of pharmaceutical preparations containing Asparagi Radix.
Résumé
Objective To control the quality of the species in Dioscorea L. better. Methods An HPLC-ELSD method was developed for the first time to simultaneously determine four bioactive ingredients:dioscin gracillin,protoneodioscin, and protoneogracillin in 31 samples belonging to seven species of Dioscorea L. from different areas. The column was an Inertsil HILIC (250mmx4.6 mm,5pm). The separation was carried out with a gradient program. The mobile phase was acetonitrile-water at a flow rate of 0.8 mL/min. Results The standard curve was rectilinear in the range of 0.464-12.97 gg (r=0.9969) for dioscin, 0.310-7.09 ltg (r = 0.9953) for gracillin, 0.469-11.66 gg (r=0.9970) for protoneodioscin, and 0.276-6.87 gg (r＝0.9992) for protoneogracillin. The recoveries of the markers were 98.1%, 100.1%, 97.2%, and 96.4%, respectively. The contents of the four components were quite different among the seven species of Dioscorea L. Conclusion The proposed HPLC-ELSD method is convenient,fast, accurate, and applicable for simultaneous analysis of multiple bioactive components of species in Dioscorea L.for quality control, which could facilitate discovering new natural resources of steroidal saponin.