SPDS Set 1 ERP Book

CHON-002

Capillary zone electrophoresis (CZE) was used for the analysis of unsaturated CS disaccharides. Response factors were used for the disaccharides (Di-0S, Di-4S, and Di-6S), for quantitation of disaccharides in the samples. CS was calculated as the amount of unsaturated disaccharides released by chondroitinase ABC divided by the ratio of 1.036 (ratio of MW of disaccharides and MW of disaccharide unit in CS). The method was run ay 60C to avoid peak splitting of the disaccharides. Size exclusion chromatography (SEC) was used to analyze CS using an RI detector. Dextrans (MW: 10K, 20K, 40K, 70K, and 150K g/mol) were used for calibration standards. The relationship of the respective dextran and log MW was evaluated. The separation was carried out using 3 columns in series. Chondroitinase ABC was used to determine the composition and content of the disaccharides. The authors state they followed a published method with modifications. The authors state the digestion time and inactivation method used was sufficient but did not provide any supporting data. The mixtures were then analyzed using CZE. Safety Review The range of the method was not defined by the authors. The authors used statistical regression for linearity. Calibration curve was evaluated for CS-A (20-200 mg/L). Routine samples were evaluated from 20-100 mg/L. There is no mention of why the curve range was varied. Calculated as 3 times the LOD. LOD was calculated from the calibration equation. LOD corresponds to the concentration of CS which gives the minimum detectable zone length equal to one second. The authors report LOQ at 9mg/mL, I calculated the LOQ at 8 mg/mL based on the information given by the authors. 8 mg/mL is at the 1% level. Its unclear if the LOD concentration was properly evaluated since the calibration curve used to extrapolate LOD concentration was much higher (20-200 mg/L) than the lowest standard concentration (LOD estimate implied at 3 mg/L). P. 59 Assessed by the average of three replicates of sample spiked with CS-A at the 50% and 100% level. 20 tablets composited, weight equivalent to one tablet was used. Accuracy was reported as 99.7 +/- 2.6% (97.1 – 102.3); therefore, not meeting the SMPR requirement of 98-103% for >10% w/w. Assessed by determination of six replicates in two different samples. 20 tablets composited, weight equivalent to one

Analytical Range:

LOQ:

Accuracy/Recovery

Precision (RSDr)