SPDS Set 1 ERP Book

CHON-002

• Difficult to determine appropriate test conditions for unknown samples.

HPLCs/UPLCs are common instruments in many labs. CE is not as widely used. Method specific drawbacks: • The method is non-specific. CS-A, B, and C had almost identical mobilities, Figure 3, p 61; therefore could not be separated using cITP. • The method is not able to differentiate CS-B (dermatan sulfate). • Acesulfam K and a high content of hydrolyzed collagen were reported interferences. The author state that Acesulfam K was easily detected in the UV trace at 254 nm. • Carrageenan and Hyaluronic could cause interference with this type of methodology. • Author stated the presence of hyaluronate and heparin did not disturb the analysis of CS but they did not provide any data. The assumption was that hyaluronate has a lower mobility than the terminator (citrate) and the heparin has a higher mobility. • In order to maximized separation multiple pH requirements are required for interfering compounds. Supporting Data o Sample preparation for the validation was one tablet dissolved in 250mL MilliQ water. One tablet, or capsule, or daily dose was used to prepare the test samples. However, the validation used a composite of 20 tablets for homogeneity reasons. There is no validation data for other dosage forms. o For the validation, external calibration solutions of CS-A were prepared at 20 – 200 mg/L to create a calibration curve using step length vs. concentration. It is unclear there was other work performed for total CS. The authors provided supplemental information: Quality Control of Chondroitin Sulphate used in Dietary Supplements. Czech Journal of Food Science 33:165-173. Modifications to the method were pH and the leading electrolyte composition. The scope of method was for raw material. Reference standards were EP CRS grade. cITP was used to determine the free sulphate content in CS. This method is unable to differentiate CS-B (dermatan sulphate)

General Comments

Method Optimization