Research On Quality Control Methods Of Ophiopogonis Radix And Seedling

Radix Ophiopogonis (RO) is the root tuber of Ophiopogon japonicus (L. f) Ker-Gawl., which has the functions of nourishing yin, producing body fluid, moistening the lung and clearing away heart-fire. In Chinese Pharmacopoeia (2010 edition), there are 95 preparations with RO which accounts for 8.9% of the total number of preparations. However, the item of RO contains the TLC identification of the standard sample and assay of total steroid saponin by spectrophotometry. RO is widely used in clinic medicine and preparation production, whose quality is mainly decided by Ophiopogonis seedling. However, breeding selection and promotion develop slowly, which will influence the development of the related industries of RO.Objective:With the methods of TLC, HPLC-ELSD and spectrophotometry, the root tubers and seedlings as the object of study, this paper has the TLC identifications and the assays of steroid saponin and steroidal sapogenin of RO, and total steroid saponins and total flavonoids of Ophiopogonis seedling, all of which has provided the scientific basis for the promotion and establishment of quality control for Ophiopogonis radix and seedling.Method:1. To determine Ophiopogonin D'of RO by both TLC and HPLC-ELSD, and to carry out the content of total steroid saponin, ash, water content and extractives followed as directed in Chinese Pharmacopoeia (2010 edition).2. To identify diosgenin and ruscogenin of RO and determine the previous one by HPLC-ELSD.3. To both determine total steroid saponins and total flavonoids of Ophiopogonis seedling by spectrophotometry.Result:1. The TLC identifications of ophiopogonin D', diosgenin and ruscogenin in RO were fulfilled.2. The determination of ophiopogonin D'and diosgenin of RO were performed by HPLC-ELSD. The linear range of ophiopogonin D'was 0.25-2.27μg, r=0.9991; and the linear range of diosgenin was 1.31-7.88μg, r=0.9991. Additionally, the results of the content of total steroid saponin, ash, water content and extractives conformed to the requirements of Pharmacopoeia.3. By means of determining the content of total steroid saponins and total flavonoids in Ophiopogonis seedling by spectrophotometry, the linear range of ophiopogonin D'was 0.071～0.152mg (r=0.9994) and the one of hesperidin was 0.010～0.033 mg·ml-1(r=0.9992).Conclusion:1. The methods of the TLC identification of ophiopogonin D'were convenient and accurate, and the assays of ophiopogonin D'and diosgenin of RO were established, which could be used to explore the methods to perfect quality contral of RO.2.In reference to Ophiopogonis seedling, both the TLC identification of diosgenin and ruscogenin and the assays of total steroid saponins and total flavonoids were established, which could provide the basis for quality contral of Ophiopogonis seedling.