title: Suppression of peak tailing of phosphate prodrugs in reversed-phase liquid chromatography.
abstract: Peak tailing of phosphate prodrugs in acidic mobile phases was thoroughly investigated. The results indicated that both metal-phosphate interactions and silanophilic interactions contributed to the observed peak tailing. Column pretreatment with phosphate buffers was demonstrated to be an effective and robust approach in suppressing metal-phosphate interaction. Silanophilic interactions, such as hydrogen bonding interactions between protonated isolated silanol groups and partially deprotonated phosphate groups were mobile phase pH dependent. The combination of column pretreatment and volatile low pH mobile phase buffers can be used to mitigate peak tailing issues in developing MS compatible RPLC methods for phosphate prodrugs. The use of non-endcapped columns should be avoided in RPLC analysis for phosphate prodrugs due to large amount of residual silanol groups in the stationary phases.
authors.affiliation: Analytical and Bioanalytical Development, Research and Development, Bristol-Myers Squibb Company, New Brunswick, NJ 08903, United States. Electronic address: Jin.zhang@bms.com.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Analytical and Bioanalytical Development, Research and Development, Bristol-Myers Squibb Company, New Brunswick, NJ 08903, United States.
authors.forename: Qinggang
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Analytical and Bioanalytical Development, Research and Development, Bristol-Myers Squibb Company, New Brunswick, NJ 08903, United States.
authors.forename: Brent
authors.initials: B
authors.lastname: Kleintop
authors.affiliation: Analytical and Bioanalytical Development, Research and Development, Bristol-Myers Squibb Company, New Brunswick, NJ 08903, United States.
authors.forename: Thomas
authors.initials: T
authors.lastname: Raglione
mesh_terms: D002021:Buffers; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods; D006863:Hydrogen-Ion Concentration; D010710:Phosphates / Q000737:chemistry*; D011355:Prodrugs / Q000737:chemistry*; D012821:Silanes / Q000737:chemistry
chemical_list: D002021:Buffers; D010710:Phosphates; D011355:Prodrugs; D012821:Silanes; C082343:silanol
keywords: HPLC mobile phase pH; Metal–phosphate interaction; Peak tailing; Phosphate prodrug; Silanophilic interaction