The determination of phosphorus in Haemophilus influenzae type b conjugate vaccines by inductively coupled plasma-atomic emission spectrometry.
This study describes a method for the determination of phosphorus in lyophilized Haemophilus influenzae type b conjugate vaccines by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The concentration of polysaccharide is directly related to the concentration of phosphorus as measured in the laboratory. Phosphorus is present in the polyribosyl-ribitol phosphate (PRP) group of the Haemophilus influenzae type b conjugate vaccine. The repeating unit of PRP is 3-B-D ribose[1-1]ribitol-5-phosphate. Phosphorus in the final container is measured in microg per dose. The amount of PRP is calculated from this and reported in microg per dose. The Haemophilus influenzae type b conjugate vaccine was analyzed for phosphorus content within the range of 1.34 to 2.02 microg phosphorus per ml. The relative difference of phosphorus concentrations determined by the ICP-AES method from the phosphorus concentrations determined by the traditional colorimetric molybdate method ranged from 2.2 to 10.6%. Phosphorus spike recovery for the vaccine ranged from 93 to 99% (1.93+/-0.13 microg P/ml). The phosphorus determination of NIST SRM 3139 phosphorus spectrometric solution differed by 3.0% from the certified phosphorus value (10.00 mg P/ml).
Center for Biologics Evaluation and Research, United States Food and Drug Administration, Rockville, MD 20852, USA.
L A
LA
Swartz
J J
JJ
Progar
J C
JC
May
D016667:Bacterial Capsules; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation / Q000379:methods / Q000592:standards; D018073:Haemophilus Vaccines / Q000737:chemistry*; D006801:Humans; D008982:Molybdenum; D010758:Phosphorus / Q000032:analysis*; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D012015:Reference Standards; D013054:Spectrophotometry, Atomic / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D018074:Vaccines, Conjugate / Q000737:chemistry
D018073:Haemophilus Vaccines; C044237:Haemophilus influenzae type b polysaccharide vaccine; D011135:Polysaccharides, Bacterial; D018074:Vaccines, Conjugate; C044659:molybdate; D010758:Phosphorus; D008982:Molybdenum