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@#This study aimed to isolate and prepare highly purified impurity C from doxycycline hyclate by a preparative HPLC method and to inspect the toxicity and in vitro antimicrobial activity of the impurity C of doxycycline hyclate. The solution of doxycycline hyclate treated with heat produced a solution containing 10% of impurity C which was firstly separated by the Sapphire C18(21. 2 mm×250 mm, 5 μm)column with 0. 1% acetic acid-acetonitrile(83 ∶17)as the mobile phase at 20 mL/min. Secondly, rotary evaporation of the eluted solution at the time of 8. 4 min was performed at 50 °C to remove organic solvent. Then the target product was prepared after freeze drying of evaporated solution adjusting pH to 1. 8 with formic acid. The target product was identified with ultraviolet absorbance(UV), infrared(IR), mass spectrometry(MS)and nuclear magnetic resonance(NMR), and its purity was be determined by HPLC. Meanwhile, cytotoxicity and genotoxicity in the Chinese hamster lung cells, toxicity on the development of zebrafish embryos and in vitro antimicrobial activity were compared among impurity C of doxycycline hyclate, doxycycline, metacycline and β-doxycycline. Results showed that prepared product was confirmed to be the impurity C of doxycycline hyclate. Its purity was 90. 1%, which had been the highest so far. In the cellular toxic tests and genetic toxic tests of Chinese hamster lung cells, impurity C of doxycycline hyclate, doxycycline, metacycline and β-doxycycline were somewhat toxic to Chinese hamster lung cells. Toxicity gradually decreased from doxycycline, impurity C of doxycycline hyclate, β-doxycycline to metacycline from -S9mix test results; toxicity gradually decreased from doxycycline, β-doxycycline, impurity C of doxycycline hyclate to metacycline from +S9mix test results; the aberration rate of all the tested related substances was less than 5%, and no obvious genotoxicity was found. According to test results of the development of zebrafish embryos, impurity C of doxycycline hyclate showed the strongest teratogenicity and lethality. Invitro antimicrobial tests revealed that impurity C of doxycycline hyclate had a weaker antimicrobial activity, and invitro antimicrobial activity potential of the tested compounds followed the order: metacycline, doxycycline, impurity C of doxycycline hyclate, β-doxycycline. Studies on safety and effectiveness indicated that impurity C of doxycycline hyclate belonged to toxic and ineffective impurity and need to be controlled individually in quality standard. A useful suggestion was given to revise the quality standard of doxycycline hyclate and its preparation in the current Pharmacopoeia of the People′s Republic of China.
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Objective To establish an ion chromatography method to determine the content of galactosamine in heparin sodium sample. Methods The content of galactosamine was determined by the ratio of response value of galactosamine and glucosamine.The determination was performed on an Dionex ICS, and the separation was carried out on a Amino acids capture column (30 mm ×3 mm), Series protect column (30 mm × 3 mm)and analytical column CarboPac PA20 (150 mm ×3 mm).The mobile phase was 14 mM potassium hydroxide solution at a flow rate of 0.4 mL/min; the column tempertature was at 30℃; the injection volume was 10μL.Results Glucosamine hydrochloride had good linearity within the range of 1.013 -16.211μg/mL(Y=2.303 4X+0.824 2,r=0.998 3), the average accuracy was 92.7%, and RSD was 3.2%(n=9), the limit of detection was 0.101 3μg/mL, and the limit of quantitation was 0.337 7μg/mL.D-Galactosamine hydrochloride had good linearity within the range of 0.010 2 -0.162 5 g/mL, (Y=31.157X-0.114 4,r=0.999 3).The accuracy was 102.1%, RSD was 2.4%(n=9).The limit of detection was 0.001 0μg/mL, and the limit of quantitation was 0.003 4μg/mL.The determination of galactosamine in 3 batches of heparin sodium raw material was not detected, (0.02 ±2.1)%, (0.03 ±1.5)%, respectively, which were all lower than the limit value (1%) of United States Pharmacopeia regulation.Conclusion The method for the determination of galactosamine in total hexose amine is successfully developed , which could be used as reference for improvement of the quality standard of heparin sodium.
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<p><b>OBJECTIVE</b>To investigate the pharmacokinetics profile of fangchinoline and tetrandrine in rats after administration of single compound and mixture with other intergradient in traditional prescription.</p><p><b>METHOD</b>A method for determination of fangchinoline and tetrandrine in rat plasma by using HPLC-MS has been developed and validated. The pharmacokinetics of two compounds and two compounds in the effective component group (ECG) of Xiaoxuming decoction were compared.</p><p><b>RESULT</b>Compared with the single dose of compound experiment results, the t(max) of fangchinoline and tetrandrine were longer than those in the single dose of ECG experiment. At the meanwhile the rest parameter showed no significant difference.</p><p><b>CONCLUSION</b>Other components in the ECG of Xiaoxuming decoction delayed the absorption rate of fangchinoline and tetrandrine, the bioavailability of two compounds were the same as that of the single dose of compound experiment.</p>
Subject(s)
Animals , Rats , Benzylisoquinolines , Pharmacokinetics , Drugs, Chinese Herbal , Pharmacokinetics , Rats, WistarABSTRACT
Objective To establish the analytical method for fingerprint of Aristolochia manshuriensis by HPLC-DAD-ESI/MS,which can be used as the basis for quality control of the drug and for the further studies on kidney toxicity metabolite.Methods Samples A.manshuriensis from different habitats were extracted by 75% methanol and analyzed by HPLC-DAD-ESI/MS,whose chromatographic fingerprints were established.Two ways to calculate the similarity were selected to compare the results by determining the common peaks.Results There were 30 main characteristic components in A.manshuriensis.The HPLC-DAD-ESI/MS fingerprint of the 30 common peaks was established preliminarily.The samples of A.manshuriensis from different habitats was found having a good similarity,and the range of similarities for 24 balches of A.manshuriensis were 0.871—0.998.Conclusion The method is reliable,accurate,and of good stability,and can be used for the quality control and variety identification of A.manshuriensis.