2013 14 4 255 3. 2 1 2 S. 2010 3 J. 2010 23 2 249. 3. 3 3% J 5 J S. 168... 2010 133.. HPLC 4.. 2010 26 6 536-537... 2010 24 3 234-235. 2013-01-15 1 2 1 1 1 2 2 311231 2* 1. 310011 2. RSD 3. 5% RSD 3. 6% 60. 8% P < 0. 001 RSD 1. 9% 8 h 60. 5% P < 0. 001 RSD 3. 1% R921. 2 A 1009-3656 2013-4 - 255-5 Zebrafish-Based Biological Quality Control for Lipid Lowering Shanqingzhi Tablets Guo Dianwu 1 Zhou Juan 2 Tang Like 1 Wu Yaodong 1 Peng Zhonghua 1 Chen Rujia 2 Zhang Yong 2 Li Chunqi 2* 1. Minsheng Pharma Hangzhou 310011 2. Hunter Biotechnology Inc Hangzhou 311231 Abstract Objective To establish larval zebrafish-based biologically quality control standard for lipid lowering traditional chinese medicine shanqingzhi tablets. Methods Biological activity / pharmaceutical effect and stability of shanqingzhi tablets were quantitatively assessed in a well-validated larval zebrafish hyperlipidemia model induced by feeding standardized high-fat food. Results Larval zebrafish hyperlipidemia model as a biological quality control standard for lipid lowering shanqingzhi tablets exhibited excellent reproducibility RSD = 3. 5% and stability RSD = 3. 6%. The preparations from a variety of batches demonstrated similar pharmaceutical activity that reduced blood lipid in larval zebrafish by 60. 8% RSD = 1. 9% P < 0. 001 after 40 hrs of drug treatment. The same batch of preparations were all stable for at least 8 hrs in room temperature and decreased zebrafish blood lipid by 60. 5% RSD = 3. 1% P < 0. 001. Conclusion Zebrafish hyperlipidemia model-based biological quality control for lipid lowering shanqingzhi tablets is a rapid reliable stable and reproducible method. Developing biological quality control standards for natural products may contribute Tel 18857129628 E-mail jackli@ zhunter. com
2013 14 4 256 Drug Standards of China 2013 Vol. 14 No. 4 to the modernization of the traditional chinese medicines. Key words zebrafish hyperlipidemia shanqingzhi tablets traditional chinese medicine quality control biology activity assay T09L901 T09L902 T09L903 1 2 Albino 6 dpf 48 h 9 O 40 h 30 0. 75% DMSO 0. 08 μg ml - 1 MESAB 2-3 O 10 4-8 IPP S 1 1. 1 % = 1 - S Albino S 100% 4 ~ 5 200 ~ 300 6 h 6 hpf 24 hpf 28 3 1L 200 mg 3. 1 480 ~ 510 μs cm - 1 ph 6. 9 ~ 7. 2 53. 7 ~ 71. 6 μg ml - 1 CaCO 3 O 9 d 1. 2 SMZ645 Nikon IPP6. 0 Media Cybernetics Nest Biotech 6 20110506 1. 3 MESAB Sigma 20110216 Methyl cellulose Sigma 20110521 20110612 O Oil Red Sigma 20110411 4% PFA 20110510 1 2-20110323 DMSO Dunnett' s T- P < 0. 05 > 90% 1 1095515 Lovastatin 1 O 20111002 > 99% A. B.
2013 14 4 257 3. 2 3. 2. 1 5 12. 5 μg ml - 1 5 7. 5 10 12. 5 15μg ml - 1 24 ~ 48 h 5 ~ 12. 5 μg P < 0. 001 ml - 1 P < 0. 001 2 4 40 h 12. 5 μg ml - 1 66. 5% P < 0. 001 3 53. 1% 48 h 66. 9% 15. 0 μg ml - 1 40 h 12. 5 P > 0. 05 μg ml - 1 P > 0. 05 12. 5 μg ml - 1 24 h 32 h 40 h 48 h 40 h 4 12. 5 μg ml - 1 x 珋 ± s n = 10 compared with Vehicle * P < 0. 001 NS 3. 3 6 40 h 6 2 12. 5 μg ml - 1 P > 0. 05 RSD 3. 5% 1 A. B. C. 12. 5 μg ml - 1 D. 0. 08 μg ml - 1 3. 4 3 40 h 3 P < 0. 001 P > 0. 05 RSD 3. 6% 2 3 1 x 珋 ± s n = 10 compared with Vehicle * P < 0. 001 3. 2. 2 4 1 2 3 4 5 6 /% 57. 6 59. 8 58. 3 56. 9 58. 7 62. 8 Mean ± SD 59. 0 ± 2. 1 RSD /% 3. 5
2013 14 4 258 Drug Standards of China 2013 Vol. 14 No. 4 2 AL20120509 AL20120512 AL20120515 /% 58. 4 61. 3 58. 5 Mean ± SD 59. 4 ± 2. 2 RSD /% 3. 6 3. 5 3. 5. 1 12. 5μg ml - 1 25 0 2 4 6 8 h 63. 2% 61. 2% 58. 2% 60. 4% 59. 4% RSD 3. 1% P < 0. 001 P > 0. 05 25 8 h 5 3. 5. 2 3 T09L901 T09L902 T09L903 12. 5 μg ml - 1 1 61. 0% 59. 2% 62. 3% P < 0. 001 P > 0. 05 RSD 1. 9% 4 4. 1 1 J 10-11 2012 13 1 5-8. 12-16 2 J 2009 7 1 1. 3 2011 2 2 59-62. 4 J. RSD 3. 5% RSD 3. 6% 4. 2 60. 8% P < 0. 001 RSD 1. 9% 8h 60. 5% P < Jersey 219-232. 0. 001 RSD 3. 1% 4. 3..... J.. 2012 26 3 224-228. 5 McGrath P Li CQ. Zebrafish a predictive model for assessing drug-induced toxicity J. Drug Discover Today 2008 13 9 394-401. 6 Li CQ Luo LQ Awerman J McGrath P. 2012. Whole Zebrafish Cytochrome P450 Assays for Assessing Drug Metabolism and Safety. Zebrafish Methods for Assessing Drug Safety and Toxicity. McGrath P ed. Wiley Press New Jersey 103-115. 7 Li CQ Luo LQ McGrath P. Zebrafish Xenotransplant Cancer Model for Drug Screening. Zebrafish Methods for Assessing Drug Safety and Toxicity. McGrath P ed. Wiley Press 2012 New 8 Li C. Q Seng W L. Park D & McGrath P. 2011. Methods for Assessing Neurotoxicity in Zebrafish. Phylonix
Cambridge MA USA. In P. McGrath Ed. Zebrafish methods for assessing drug safety and toxicity pp. 117-134. West Sussex UK Wiley-Balckwell. 9 Clifton JD Lucumi E Myers MC Napper A Hama K et al. I- dentification of Novel Inhibitors of Dietary Lipid Absorption Using Zebrafish J. PLoS ONE 2010 5 8 1-9. 10 Parng C Seng WL Semino C McGrath P. Zebrafish a preclinical model for drug screening. Assay Drug Dev Technol 2002 1 41-48. 11 Parng C. In vivo zebrafish assays for toxicity testing. Curr Opin Drug Discov Dev 2005 8 100-106. 12 Sheridan MA. Lipid dynamics in fish aspects of absorption transportation deposition and mobilization. Comp Biochem Physiol B 1986 90 679-90. 2013 14 4 259 13 Schlegel A Stainier DY. Microsomal triglyceride transfer protein is required for yolk lipid utilization and absorption of dietary lipids in zebrafish larvae. Biochemistry 2006 45 15179-15187. 14 Farber SA Pack M Ho SY Johnson ID Wagner DS et al. Genetic analysis of digestive physiology using fluorescent phospholipid reporters. Science 2001 292 1385-1388. 15 Kadereit B Kumar P Wang WJ Miranda D Snapp EL Severina N et al. Evolutionarily conserved gene family important for fat storage. Proc Natl Acad Sci U S A 2008 105 94-99. 16 Stoletov K Fang L Choi SH Hartvigsen K Hansen LF Hall C et al. Vascular lipid accumulation lipoprotein oxidation and macrophage lipid uptake in hypercholesterolemic zebrafish. Circ Res. 2009 104 952-60. 2012-10-11 HPLC-RID 430064 - NH 2 - HPLC-RID 1. 0 ml 2000 ml ph 7. 5-45 55 0. 6 ml min - 1 10 ~ 320 μg r = 1. 0000 99. 7% RSD 0. 32% n = 9 R921. 2 A 1009-3656 2013-4 - 259-5 Determination of the Content of Glucosamine Hydrochloride by HPLC-RID Jiang Hong Dan Xiaomeng Hu Yuanhua Xu Xiaoling Hubei Institute for Food and Drug Control. Wuhan 430064 Abstract Objective To establish an HPLC-RID method for determining the content of glucosamine. Methods The NH 2 column was used and mobile phase was ammonium phosphate buffer dissolve phosphate 1. 0 ml into 2000 ml water adjust the ph to 7. 5 using ammonia solution -acetonitrile 45 55. The flow rate was 0. 6 ml min - 1. Results A good linear range of glucosamine hydrochloride was 10-320 μg r = 1. 0000 the average recovery was 99. 7% RSD = 0. 32% n = 9. Conclusion The method is simple accurate and reliable. It is suitable for quantity control of glucosamine. Key words HPLC RID glucosamine dydrochloride content determination 1-3 4-7 Tel 15697185050 E-mail hbyjsjh@ 163. com