| Fortune’s Drynaria Rhizome is the dried rhizome of Drynaria fortunei (Kunze) J. Sm., which can replenish the kidney, strengthen the bones, promote the healing fracture and relieve pain. In order to control the quality and ensure the security and efficiency in clinical use of Fortune’s Drynaria Rhizome, the quality standard of Fortune’s Drynaria Rhizome was developed and its pharmacokinetics was studied.The method of fingerprint analysis and quantification of Rhizoma Drynariae was performed by high performance liquid chromatography on Diamonsil C18 column. The column was eluted with acetonitfile-0.4% glacial acetic acid at 25℃and 254 nm. Fingerprint analysis of Rhizoma Drynariae using HPLC profiling was established for both ethanol and cyclohexane extracts. The contents of naringin, neoeriocitrin and (E)-4-O-β-D-glucopyranosyl caffeic acid in nineteen batches were determined simultaneously. Significant differences were observed between genuine and fake samples in the principal components analysis (PCA) score plot, finding four markers in the PCA loading plot as well. Three important markers were naringin, neoeriocitrin and (E)-4-O-β-D-glucopyranosyl caffeic acid, and the contents of them in ten genuine batches were 6.36～10.1 mg/g,5.14～9.21 mg/g and 1.87～3.19 mg/g, respectively. Combining determination of the index components with fingerprint analysis of Rhizoma Drynariae using HPLC profiling, the quality of Rhizoma Drynariae can be assessed better.We developed the Gas chromatographic fingerprint of Rhizoma Drynariae. The samples were placed in a Soxhlet’s extractor with cyclohexane and refluxed for 3h. The GC conditions were as follows:a DB-5 column, programmed column temperature, FID detector. Significant differences were observed between genuine and fake samples in the PCA score plot. Twenty commom peaks were identified by GC-MS. This method provides useful reference for quality control of Rhizoma Drynariae.A rapid, specific and sensitive LC-MS/MS method was developed for the determination of naringin and naringen in rat plasma. The analysis was carried out on an ACQUITY UPLCTM EH C18 column(100 mm×2.1 mm, i.d.,1.7μm) by isocratic elution with acenitrile-0.4% acetic acid (80:20, v/v) as mobile phase at a flow rate of 0.25 ml/min. Paracetamol was used as the internal standard. Detection was carried out by means of negative electrospray ionization (ESI) mass spectrometry with multiple-reaction monitoring (MRM). Quantification was performed using the transitions of m/z 578.4â†'270.7 for naringin, m/z 270.7â†'150.7 for naringenin and m/z 149.8â†'106.6 for paracetamol, respectively. Linear calibration curves of naringin and naringenin were obtained over the concentration ranges of 5.00～5.00×103 ng/ml and 2.00～400 ng/ml, with a low limit of quantification of 5.00 ng/ml and 2.00 ng/ml, respectively. The matrix effect was minimized. The intra- and inter-day precisions(RSD) were within 6.9% and 8.1% for naringin and 10.0% and 12.6% for naringenin. The accuracy(RE) was from 0.7 to 3.2% for naringin and from -0.6 to 0.9% for naringenin. The validated method was successfully applied to the pharmacokinetic studies of naringin and naringenin in rats after oral administration of Rhizoma Drynariae extract.The Cmax of naringin and naringenin were (2.56±0.77)×103 ng/ml and 222±45 ng/ml, the Tmax were 0.67±0.20 h and 8.0±1.3 h, the t1/2 were 4.1±0.8 h and 3.5±0.8 h, and the AUC0～∞were (3.29±0.54)×103 ng/ml-h and (1.79±0.43)×103 ng/ml-h, respectively. Compared with naringenin, naringin appeared to be absorbed fast and eliminated slowly. |
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