Direct comparison of LC–MS/MS and RIA methods for the pharmacokinetics assessment of human insulin in preclinical development

Abstract

Insulin is an effective therapeutic for diabetes, and the level of insulin in vivo is directly related to the health of diabetic patients. Traditionally, the concentrations of insulin in vivo are determined by the radioimmunoassay (RIA) method. In this study, we developed an LC-MS/MS method for the quantification of human insulin in dog plasma and directly compared the RIA and LC-MS/MS methods. Our LC-MS/MS method exhibited superior accuracy, efficiency and cost-effective for the pharmacokinetic (PK) assessment of human insulin. The LC-MS/MS method can quantitate human insulin and canine insulin simultaneously without cross-reactivity, making the analysis more efficient. The LLOQ of our LC-MS/MS method was 38.5 pg/mL, which was necessary to fully describe the PK profiles of endogenous and exogenous insulin in vivo. The direct comparison of PK data obtained from the two methods demonstrated that LC-MS/MS could be an alternative to the RIA method and should be widely used for the quantification of insulin drugs, especially in preclinical studies.

Timapiprant, a prostaglandin D2 receptor antagonist, ameliorates pathology in a rat Alzheimer’s model

“…This discrepancy can be attributed to the different methodology used, radioimmunoassays in the older studies ( Narumiya et al, 1982 ) versus quantitative LC–MS/MS in the most recent studies ( Shaik et al, 2014 ) and our studies. LC–MS/MS exhibits superior sensitivity, accuracy, efficiency, and lack of cross-reactivity compared with radioimmunoassays ( Brose et al, 2011 , 2013 ; Dong et al, 2018 ).…”

Targeted and Untargeted Proteomics Approaches in Biomarker Development

“…It is time‐consuming and costly to develop a new radio‐immunoassay method for the quantification of every new insulin analog. In contrast, developing LC–MS/MS methods for insulin quantification is more straightforward, efficient, and therefore has become common in recent years . For example, Chambers et al.…”

“…developed an LC–MS/MS method capable of differentiating and simultaneously quantifying human insulin and the therapeutic insulin Glargine. They demonstrated that their LC–MS/MS method was more accurate and efficient for the assessment of human insulin pharmacokinetics than a radio‐immunoassay method …”

Simplified quantification of insulin, its synthetic analogs and C‐peptide in human plasma by means of LC‐HRMS

“…[4][5][6][7][8][9] Other assays using solid phase extraction strongly focus on insulins (without C-peptide) only. [10][11][12] The immune-enriched sample aliquots are of highly purified quality, enabling the injection into nano-scale liquid chromatographs without the risk of blocking due to residual matrix components. The accomplished specificity of these assays were surpassing, but in terms of quantification, several issues may arise: Due to the limited capacity of the antibodies, the linear range of antibody-based assays is restricted and in the case of co-existing anti-insulin antibodies, which were occasionally described in patients with diabetes receiving exogenous insulin therapy, the extraction by means of techniques employing immunoaffinity interaction is potentially affected.…”

“…Existing liquid chromatography/mass spectrometry‐based assays mainly use immunoaffinity‐assisted extraction or other sophisticated isolation techniques for the target peptides from the matrix . Other assays using solid phase extraction strongly focus on insulins (without C‐peptide) only . The immune‐enriched sample aliquots are of highly purified quality, enabling the injection into nano‐scale liquid chromatographs without the risk of blocking due to residual matrix components.…”