1-4 certified reference material CRM. DSC HPLC ± 0. 4 % k = 2 P = GBW
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1 * DSC HPLC ± 0. 4 % k 2 P GBW09587 R R A Purity Determination and Uncertainty Evaluation of Ribavirin Purity Reference Material DU Hui 1 XING Cheng 1 SHI Yong-qiang 2 DU Guan-hua 3 LV Yang Beijing City Key Laboratory of Pharmaceutical Polymorph Institute of Materia Medica Chinese Academy of Medical Sciences & Peking Union Medical College Beijing China 2. Shandong Province Key Laboratory of Pharmaceutical Polymorph Tengzhou China 3. Beijing City Key Laboratory of Drug Identification and Drug Screening Institute of Materia Medica Chinese Academy of Medical Sciences & Peking Union Medical College Beijing China ABSTRACT Objective To develop the ribavirin purity certified reference material CRM which has measurement traceability and high accuracy and establish an effective method of evaluation. Methods The homogeneity and the stability were checked by differential scanning calorimetry DSC. High performance liquid chromatography HPLC and DSC methods were used for purity determination of ribavirin. Results Ribavirin showed satisfactory homogeneity and stability. The certified value of ribavirin was 99. 5% with an uncertainty of 0. 4% k 2 P Conclusion Ribavirin purity CRM obtained in this paper has been proven to be a national primary CRM with high accuracy and traceability which can be used to validate analytical methods improve the accuracy of measurement data establish meteorological traceability of analytical results as well as control the quality of ribavirin in the pharmaceutical industry. KEY WORDS Ribavirin Purity certified reference material Calorimetry differential scanning Chromatography high performance liquid 1-4 certified reference material CRM imm. ac. cn * 5-7 CRM 2012ZX ZX com dugh@ imm. ac. cn
2 780 Herald of Medicine Vol. 33 No. 6 June differential scanning calorimetry DSC Mettler Toledo DSC-1 Al 2 O 3 DSC 40 μl Al 2 O K min ~ 180 chromatography HPLC Agilent HPLC DAD Agilent Eclipse XDB-C mm 4. 6 mm 5 μm ph % 207 nm ml 1. 0 ml min μl μg ml Mettler Toledo V20 SX ml 10 ml Agilent A μg ml - 1 J. T. Baker g 500 ml 24 h h mg HPLC 25 ml 98. 0% μg ml μl > 99. 0% 6 2 ml mg RSD 0. 07% HPLC DSC ~ 3. 5 mg mg 40 μl Al 2 O 3 DSC RSD 0. 42% HPLC 1. 5 ~ 3. 5 mg 2. 2 Y X R mg 40 μl ~ 3. 5 mg RSD 1. 53% DSC high performance liquid mg μl GBW ± K Millipore ~ μg ml μm Y X R n 6 HPLC mg 25 ml μg ml μl n 6 DSC 15 3 DSC
3 F F 1. 44F F F < F t t t DSC t t t t % ± 5% t t ± 500 lx d Tab. 1 1 Results of homogeneity test on purity certified reference material of ribavirin Y ij /% n i N Y i i j j Y j N Y 2i j i j s i Tab. 2 / 2 Results of short-term stability test on purity certified reference material of ribavirin /% / d %
4 782 Herald of Medicine Vol. 33 No. 6 June 2014 Tab. 3 3 Results of long-term stability test on purity certified reference material of ribavirin /% / / % x y 100% y x a S 2 n Y i - b - ax i 2 /n i n P % t a < t4 S a f i A i 10 G G G G G < G G < G HPLC DSC % % A i C i 100% f i A i C i A i f i DSC DSC DSC Tab. 4 Determination results of purity certified reference material of ribavirin by DSC methods < 2% Van t Hoff / / / x 1 - T mg % mg % 0 - T m ΔH 2 100% RT T 0 3 K T m K R J K - 1 mol - 1 x H J mol - 1 T 0 - T m mg ml 500 μg ml ~ 3. 5 mg 40 μl 10 μl 3 Al 2 O HPLC HPLC HPLC %
5 Tab. 5 A 1 5 HPLC Determination results of purity certified reference material of ribavirin by HPLC methods A A 2 A A 3 A A A S S S / % u 2 B1 DSC + u 2 A1 DSC + u 2 B2 DSC + u 2 B3 DSC % HPLC HPLC % % % 100% % % DSC HPLC 6 DSC t t Tab. 6 Combined standand uncertainty results of DSC t method % % / % u B1 DSC B u 2. 4 A1 DSC A u B2 DSC B 3 u B3 DSC B u H MS 2 - MS2 /n u A S 2 / / u A S 2 / S a S u c x peak % n X i - X - 2 u ( A ) + u 2 ( A ) i 1 u t t. S a u c x H2 O s / DSC DSC u c x ash s / JJFl u c x solvent s / 10 u c x DSC /99. 49%
6 784 Herald of Medicine Vol. 33 No. 6 June 2014 HPLC u c x HPLC u c x peak 2 + u c x H2 O 2 + u c x ash u c x solvent REICHARD O ANDERSON J SCHVARCZ R et al. Ribavirin treatment for chronic hepatitis C J. Lancet u c ( ) x x - ( u c ( x ) ) 2 + ( u c ( x ) ) /x DSC DSC 99.48% /x 2 HPLC HPLC u c u 2 c x + u 2 H + u 2 t FONTINE H VALLET A SELVES J et al. Histopathologic efficacy of ribavirin monotherapy in kidney allograft k 2 recipients with chronic hepatitis C J. Transplantation U ku c ISO Guide 34 S ISO Guide 35 S % ± 0. 4% k 2 P 7 National Institute of Standards and Technology OL. http / / www. nist. gov / srm / index. cfm. 1 VINCENT M ELISABETH N PHIIPPE S. Brief communication case reports of ribavirin treatment for chronic hepatitis E J. Ann Int Med GARNIER J L CHEVALLIER P DUBERNARD J M et al. Treatment of hepatitis C virus infection with ribavirin in kidney transplant patients J. Transplant Proc DUEWER D L LIPPA K A LONG S E et al. Demonstrating the comparability of certified reference materials J. Anal Bioanal Chem COMAR DB. http / /www. comar. bam. de /en /. 10. M. GBW HPLC S. JJF ARIANNA L MARIA C S SILVANA P. Measurement of ribavirin and evaluation of its stability in human plasma by DSC HPLC high performance liquid chromatography with UV detection J. J Chromatogr B HPLC 13 ELISA Z TERESA G BARBARA L et al. Development of a rapid LC-MS / MS method ro ribavirin determination in rat DSC brain J. J Pharm Biomed Anal LIU Y XU C YAN R. Sensitive and specific LC-MS /MS DSC method for the simultaneous measurements of viramidine and ribavirin in human plasma J. J Chromatogr B DSC HPLC 15 MICHAEL C B REGULA T. Determiantion of ribavirin in human serum and plasma by capillary electrophoresis J. Electrophoresis J DOI /yydb
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