Development & Validation of Biochemical Method for estimation of Methyl Pentose content in Pneumonia Vaccine
A biochemical method was developed for determination of methyl pentose content in polysaccharide pneumonia vaccine. The estimation of methyl pentose in vaccines is important since it acts an an effective antigen. The assay development & protocol was developed & conducted at the laboratories of Serum Institute of India Ltd. over a period of 6 months.
The following protocol was followed:
The vaccine sample was hydrolyzed using Sulphuric Acid at 90 degree Celsius for 10 minutes, to release the methyl pentose residue.
The methyl group of methyl pentoses reacted with "-SH" group of cystine hydrochloride.
This reaction produces a pale yellow color.
These colored products are read on an ELISA reader.
The quantitative determination of methyl pentose residues was done by using a standard preparation of rhamnose sugar.
In order to demonstrate that this method is suitable for methyl pentose estimation, the method was validated using ICH tripartite guidelines which included parameters such as:
● Specificity
● Range
● Repeatability
● Ruggedness
● Robustness
● Solution Stability