Development of Glucosamine Sulfate Certified Reference Material

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4 015 3 CHEMICAL ANALYSIS AND METERAGE Vol. 4 No. Mar. 015 1 doi 10.3969 j.in.1008 6145.015.0.001 * 1 1 1 1 1 1 1. 361005. 361005 IRNMR 00 15 95% 5 1 8 99.84% 0.18% k=1.96 O653 A 1008 6145(015)0 0001 05 Development of Glucoamine Sulfate Certified Reference Material Chen Weizhu 1, Zhang Yiping 1, Fang Hua 1, Yi Ruizao 1, Zhu Ruilin 1, Hong Zhuan 1 (1. The Third Intitute of Oceanography of the State Oceanic Adminitration, Xiamen 361005, China. Department of Chemitry and Key Laboratory Chemical Biology of Fujian Province, Xiamen Univerity, Xiamen 361005, China) Abtract Glucoamine ulfate certified reference material wa developed. Glucoamine ulfate wa made from the glucoamine hydrochloride. The tructure of glucoamine ulfate certified reference material wa comfirmed by infrared pectrocopy (IR), high reolution ma pectral and nuclear magnetic reonance (NMR). The ample wa divided into 00 bottle, the homogeneity, tability teting and quantitative analyi were carried out by high performance liquid chromatography with evaporative light-cattering detector (HPLC ELSD). According to analyi procedure of homogeneity under the confidence interval of 95%, 15 bottle of ample were randomly taken from 00 bottle, and the reult were validated by F-tet tatitical method.the tability inpection wa carried on the long-term(1 month), and the reult indicated that the period for glucoamine ulfate of torage wa 1 month at 5. A cooperative certification wa conducted with eight qualified laboratorie. The certified purity value of the reference material of glucoamine ulfate wa 99.84% with a relative expaned uncertainty of 0.18%(k=1.96). The reference material can conform to the technical requirement of the certified reference material. The material wa intended for ue in the method validation and quality control regarding glucoamine ulfate. Keyword glucoamine ulfate; certified reference material; homogeneity; tability; certification; uncertainty 1 EOLAR * 350Z0130004 1PZP001SF10 GD01 D01 001 E-mail: hzh@tio.org.cn 015 01 16

015 4 3 1 Water Alliance Water 695 eparation Module Water 44 ELS Detector Water HP 845 Nicolet Ftir-670 Thermo Scientific Lct Premier TM XE Water AL 104 Milli-Q Milli-pore Milli Q 50 mg 100 ml 500 μg ml 10 μl 3 3 3.1 1 OH CH CHNH + 3 C O C N 1 cm 1 3 395 OH NH O H N H 1 089 1 031 CO CN SO C O C N SO 4 95 CH CH m 1 445 CH CH m 1 378 1 633 1 534 CH NH 3 + CH NH 3 + 3. C 6 H 14 NO 5 + 180.086 7 180.087 0 C 6 H 1 NO 4 + 16.076 1 16.076 3 C 1 H 7 N O 10 + 359.165 6 359.166 7 δ C (100 MHz) δ H ( J 400 MHz) 1 89.08 5.31 (d J=3.6 Hz 0.6) 1 9.66 4.81 (d J=8.4 Hz 0.4) 54. 3.16 (dd J=3.6 Hz J=10.4 Hz 0.6) 56.66.87 (dd J=8.4 Hz J=10.4 Hz 0.4) 3 69.53 3.70~3.67 (m 0.6) 3 71.90 3.41~3.38 (m 0.4) 4 69.49 3.37~3.34 (m 0.6) 4 69.61 3.33~3.09 (m 0.4) 5 71.51 3.79~3.77 (m 0.6) 5 76.08 3.59~3.54 (m 0.4) 6 60.3 3.75~3.73 (m 1.) 6 60.39 3.66~3.60 (m 0.8) (D O) δ5.31 (d J=3.6 Hz) δ4.8 (d J=8.4 Hz) 3 α β 4 α β 1 3 3 6 5 O 4 OH - 3 1 SO 4 NH 3 + α m

3 4' 3' 6' 5' O ' 1' NH + 3 OH SO 4 - β 1 α β 4 00 mg 00 GB T 15000.3 008 5 15 3 3 4 3% 1 99.76 99.77 99.77 99.77 11 99.75 99.79 99.78 99.77 1 99.81 99.79 99.79 99.80 31 99.83 99.77 99.79 99.80 41 99.8 99.74 99.76 99.77 51 99.79 99.78 99.81 99.79 61 99.79 99.80 99.83 99.81 71 99.74 99.80 99.79 99.73 81 99.74 99.75 99.75 99.75 91 99.80 99.78 99.75 99.78 101 99.86 99.79 99.76 99.80 111 99.75 99.79 99.76 99.77 11 99.78 9.78 99.79 99.78 131 99.80 99.76 99.76 99.77 141 99.80 99.74 99.76 99.77 4 SS( ) MS( ) 0.011 400 14 0.000 814 0.00 400 30 0.000 680 0.031 800 44 F 0.05 14 30 =.04 F=MS MS =1.0 F<F 0.05 14 30 95% 5 5 1 0 1 3 6 9 1 5 55 % 0 99.89 3 99.8 6 99.91 9 99.81 1 99.89 GB T 15000 008 3 u lt = b t=0.07% 1 6 6.1 6 7 8 9 10 1 13 14 6. Prevail Carbohydrate ES (50 mm 4.6 mm 5 μm) 70 30 0.8 ml min 8 80 75.8 kpa 10 L 7 7.1 GB T 15000.3 008 8 8 1 6

4 015 4 6 1 99.70 99.7 99.7 99.7 99.7 99.74 99.7 0.0 99.77 99.80 99.83 99.84 99.84 99.86 99.8 0.04 3 99.98 99.98 99.98 99.98 99.99 99.99 99.98 0.01 4 99.89 99.90 99.91 99.93 99.94 99.94 99.9 0.03 5 99.66 99.69 99.70 99.75 99.76 99.79 99.73 0.05 6 99.7 99.73 99.73 99.74 99.74 99.75 99.74 0.0 7 99.97 99.8 99.83 99.83 99.86 99.86 99.83 0.03 8 99.64 99.73 99.73 99.79 99.85 99.90 99.77 0.01 Grubb Grubb 8 Cochran (1) C C = n / m ax i= 1 i ( 1) max i i C 8 6(0.05) =0.36C=0.36 C > C 8,3(0.05) 99.84% 7. GB T 15000.3 008 U CRM U(x) U bb U lt 7 7 u char u bb u lt % 0.04 0.0 0.07 () u = u char + u bb + u lt ( ) 7 () u=0.09% U 95 =ku=0.18% k= 99.84% 95% 0.18% k= 8 99.84% 0.18% k= 1Piperno M Reboul P Hellio Le Graverand M P et al. Glucoamine ulfate modulate dyregulated activitie of human oteoarthritic chondrocyte in vitro J. Oteoarthriti and Cartilage 000(8): 07 1. Hungerford D S Jone L C Glucoamine and chondroitin ulfate are effective in the management of oteoarthriti J. The Journal of Arthroplaty 003 18(3): 5 9 3Largo R Alvarez-Soria M A Díez-Ortego I et al. Glucoamine Inhibit IL 1 beta-induced NFkappaB activation in human oteoarthritic chondrocyte J. Oteoarthriti Cartilage 003 11(4): 90 98. 4Der-Lii M T. A Solid-tate NMR Application of the anomeric effect in carbohydrate galactoamine glucoamine and n-acetylglucoamine J. Solid State Nuclear Magnetic Reonance 005 7: 09 14. 5GB T 15000.3 0083 S. 6. HPLC ELSD J. 008 7(): 41 43. 7. HPLC ELSD J. 008 3(): 9 30. 8Wang X M Zhang R L Lv Z H et al. Determination of glucoamine and lactoe in milk-baed formulae by highperformance liquid chromatography J. Journal of Food Compoition and Analyi 008 1: 55 58. 9Pun ın Crepo M O Vilaoa Mart ınez M L opez Hern andez J et al. High-performance liquid chromatographic determination of chitin in the now crab chionoecete opilio J. Journal of Chromatography A 006 1 116: 189 19. 10. HPLC J. 01 43(6): 466 467. 11. J. 009 5(3): 80 81. 1. J. 01 18(3): 40 4.

5 13. LC MS MS J. 011 (14): 1 84 1 85. 14Hubert C Houari S Lecomte F et al. Development and validation of a enitive olid phae extraction hydrophilic interaction liquid chromatography ma pectrometry method for the accurate determination of glucoamine in dog plama J. Journal of Chromatography A 010 1 17: 3 75 3 81. 36 014 5.8 5 6.9% 019 36.0 5 5.7% 019 8.84 5 5 ( ) 0.05 mg m 3 HJ543 ( ) ( ) 30% GB 7665 1987 4 GHz 5 5 00 600 70% MEMS ( ) 015 7 015 (