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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

General Comments

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.

Method Optimization

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)