Zebrafish-Based Biological Quality Control for Lipid Lowering Shanqingzhi Tablets

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1 S J % J 5 J S HPLC * RSD 3. 5% RSD 3. 6% 60. 8% P < RSD 1. 9% 8 h 60. 5% P < RSD 3. 1% R A 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 Hunter Biotechnology Inc Hangzhou 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 < 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 < 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 jackli@ zhunter. com

2 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 T09L Albino 6 dpf 48 h 9 O 40 h % DMSO μg ml - 1 MESAB 2-3 O IPP S % = 1 - S Albino S 100% 4 ~ ~ h 6 hpf 24 hpf L 200 mg ~ 510 μs cm - 1 ph 6. 9 ~ ~ μg ml - 1 CaCO 3 O 9 d 1. 2 SMZ645 Nikon IPP6. 0 Media Cybernetics Nest Biotech MESAB Sigma Methyl cellulose Sigma O Oil Red Sigma % PFA DMSO Dunnett' s T- P < > 90% Lovastatin 1 O > 99% A. B.

3 μg ml μg ml ~ 48 h 5 ~ μg P < ml - 1 P < h μg ml % P < % 48 h 66. 9% μg ml h P > μg ml - 1 P > μg ml h 32 h 40 h 48 h 40 h μg ml - 1 x 珋 ± s n = 10 compared with Vehicle * P < NS h μg ml - 1 P > RSD 3. 5% 1 A. B. C μg ml - 1 D μg ml h 3 P < P > RSD 3. 6% x 珋 ± s n = 10 compared with Vehicle * P < /% Mean ± SD ± 2. 1 RSD /% 3. 5

4 Drug Standards of China 2013 Vol. 14 No. 4 2 AL AL AL /% Mean ± SD ± 2. 2 RSD /% μg ml h 63. 2% 61. 2% 58. 2% 60. 4% 59. 4% RSD 3. 1% P < P > h T09L901 T09L902 T09L μg ml % 59. 2% 62. 3% P < P > RSD 1. 9% J J J. RSD 3. 5% RSD 3. 6% % P < RSD 1. 9% 8h 60. 5% P < Jersey RSD 3. 1% J McGrath P Li CQ. Zebrafish a predictive model for assessing drug-induced toxicity J. Drug Discover Today Li CQ Luo LQ Awerman J McGrath P 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 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 Methods for Assessing Neurotoxicity in Zebrafish. Phylonix

5 Cambridge MA USA. In P. McGrath Ed. Zebrafish methods for assessing drug safety and toxicity pp 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 Parng C Seng WL Semino C McGrath P. Zebrafish a preclinical model for drug screening. Assay Drug Dev Technol Parng C. In vivo zebrafish assays for toxicity testing. Curr Opin Drug Discov Dev Sheridan MA. Lipid dynamics in fish aspects of absorption transportation deposition and mobilization. Comp Biochem Physiol B Schlegel A Stainier DY. Microsomal triglyceride transfer protein is required for yolk lipid utilization and absorption of dietary lipids in zebrafish larvae. Biochemistry Farber SA Pack M Ho SY Johnson ID Wagner DS et al. Genetic analysis of digestive physiology using fluorescent phospholipid reporters. Science 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 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 HPLC-RID NH 2 - HPLC-RID 1. 0 ml 2000 ml ph ml min ~ 320 μg r = % RSD 0. 32% n = 9 R A 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 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 The flow rate was 0. 6 ml min - 1. Results A good linear range of glucosamine hydrochloride was μg r = 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 Tel hbyjsjh@ 163. com