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ObjectiveTo improve the current standard of Belladonnae Herba in the 2020 edition of Chinese Pharmacopoeia. MethodTaking hyoscyamine sulfate， atropine sulfate and scopoletin as reference substances， and ethyl acetate-methanol-concentrated ammonia（17∶4∶2）as developing solvent， thin layer chromatography （TLC） was applied in the qualitative identification of Belladonnae Herba. The moisture， total ash and ethanol-soluble extract of Belladonnae Herba were determined based on the general principles in the 2020 edition of Chinese Pharmacopoeia （volume Ⅳ）. The contents of hyoscyamine sulfate and scopolamine hydrobromide were analyzed by high performance liquid chromatography （HPLC） with mobile phase of acetonitrile-54 mmol·L-1 phosphate buffer solution （14∶86）， flow rate of 1.0 mL·min-1 and detection wavelength at 210 nm. ResultThe spots in the TLC were clear with good separation and specificity. Hyoscyamine sulfate and scopolamine hydrobromide showed a good linearity with peak area in the range of 0.024 7-0.789 6 g·L-1 （r=0.999 9） and 0.003 9-0.124 0 g·L-1 （r=0.999 9）， the average recoveries of these two ingredients were 100.29% （RSD 1.6%） and 99.04% （RSD 1.4%）， respectively. The limits for moisture， total ash in Belladonnae Herba should be less than 13.0% and the limit for the ethanol-soluble extract should be more than 10.0%. Due to the low content and wide variation of scopolamine hydrobromide， the content of hyoscyamine sulfate should not be less than 0.098%. ConclusionThe established method is simple， specific and reproducible， which can be used to improve the quality control standard of Belladonnae Herba.
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AIM To establish an RP-HPLC method for the simultaneous content determination of two constituents in five parts (whole grass,roots,stems,leaves and fruits) of Belladonnae Herba.METHODS The analysis of Belladonnae Herba methanol extract was carried out on a 35 ℃ thermostatic Agilent TC-C18 column (4.6 mm × 150 mm,5 μm),with the mobile phase comprising of acetonitrile-phosphate buffer flowing at 1.0 ml/min in an isocratic elution manner,and the detection wavelength was set at 216 nm.RESULTS Atropine sulfate and scopolamine hydrobromide showed good linear relationships within the ranges of 51.60-1 290 mg/L (R2 =0.999 9)and 2.90-72.00 mg/L (R2 =0.999 5),whose average recoveries were 101.3％ and 101.5％ with the RSDs of 2.5％ and 1.3％,respectively.The contents of two constituents were the highest in the roots and leaves,respectively,and both of them demostrated the lowest contents in the stems.CONCLUSION The contents of atropine sulfate and scopolamine in different parts of Belladonnae Herba show obvious differences.
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Objective: To investigate the dynamic accumulations and distributions of eight chemical compounds of Belladonnae Herba in different growing periods and explore the distribution of each ingredient in various parts. Methods: The detection was performed by RP-HPLC with diode array detector (DAD). The mobile phase was acetonitrile-0.05% phosphonic acid in gradient elution. The HPLC method was established for the simultaneous determination of the eight compounds, such as scopolin, chlorogenic acid, quercetin-3-O-galactose (6→1) rhamnose-7-O-glucoside, quercetin-3-O-glucose (6→1) rhamnose-7-O-glucoside, kaempferol-3-O-galactose (6→1) rhamnose-7-O-glucoside, kaempferol-3-O-glucose-(6→1) rhamnose-7-O-glucoside, scopoletin, and rutin, and also for the the comparison on the changes of each ingredient in different growing periods and distribution in various parts. Results: The total amount of each ingredient in the whole plant increased along with its growth. However, each ingredient had different rate of increase, in addition, most compounds would reach to a balance of accumulation in the middle or the last third of June. The analysis results showed scopolin mainly distributed in the roots, while chlorogenic acid had a higher distribution in the leaves and flowers; Scopoletin distributed evenly among all parts in the plant, and the others possessed a higher distribution among the leaves and flowers, however, there was scare distribution in the roots. Conclusion: Each ingredient of Belladonnae Herba has different rate of increase and different distribution in each part, resulting in comparatively large difference of each compound. The experimental data of this research could provide the basis for the implantation, harvest and collection, and quality evaluation of Belladonnae Herba.