Method for the quantitative determination of poloxamers
The invention relates to the analytical determination of poloxamers in a liquid protein sample.
1. A method for the quantitative determination of a poloxamer in a liquid protein sample comprising the steps of subjecting said sample to:
a) a separation step using a size exclusion chromatography column;
b) an elution step with a mobile phase adjusted to a pH below 3; and
c) detecting the amount of poloxamer in said sample.
2. The method of claim 1 , wherein the poloxamer is Poloxamer 188.
3. The method of claim 1 , wherein the protein in said liquid protein sample has a molecular mass between 5-70 kDa.
4. The method of claim 3 , wherein the protein has a molecular mass of between 20-70 kDa.
5. The method of claim 1 , wherein the protein is a heterodimer protein.
6. The method of claim 5 , wherein the protein is a gonadotropin selected from FSH, LH, hCG, TSH.
7. The method of claim 1 , wherein the protein to be analysed is interferon beta or Growth Hormone (GH).
8. The method of claim 1 , wherein the sample is an aqueous pharmaceutical composition containing FSH, LH, hCG, TSH, GH or interferon beta.
9. The method of claim 1 , wherein the mobile phase is aqueous.
10. The method of claim 1 , wherein the elution of the sample is performed with a mobile phase of acidic pH.
11. The method of claim 1 , wherein the pH of the mobile phase is adjusted to about 1.9-2.0.
12. The method of claim 1 , wherein the detection of poloxamer comprises analyzing the refractory index.
13. The method of claim 1 , wherein the size exclusion column is a SE-HPLC column filled with a polymer based matrix containing beads.
14. The method of claim 13 , wherein the beads of the matrix have a particle size of 10 or 17 μm.
15. The method of claim 13 , wherein the beads of the matrix have a pore size of about 200 Å.
16. The method of claim 14 , wherein the beads of the matrix have a pore size of about 200 Å.