Response boosting-based approach for absolute quantification of gelatin peptides using LC-MS/MS

Area of Science:

• Analytical Chemistry
• Biochemistry

Background:

• Gelatin, particularly Asini Corii Colla, is prevalent in Asian food therapy products.
• Collagen enzymolysis yields glycine residues, which can boost mass spectrometry (MS) signals.
• Accurate quantification of gelatin is crucial for quality control.

Purpose of the Study:

• To develop a novel response-boosting strategy for enhanced MS detection sensitivity.
• To achieve absolute quantification of Asini Corii Colla using high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS).

Main Methods:

• Utilized response-boosting to enhance MS signal detection.
• Employed peptidomics analysis to select an appropriate simulated matrix (deer-hide gelatin).
• Incorporated isotope-labeled internal standards and matrix-matched calibration curves for accurate quantification.
• Validated the method for three species-specific gelatin peptides.

Main Results:

• The established method demonstrated excellent linearity (r² > 0.997), precision (RSD < 8.5%), repeatability (RSD < 8.9%), and accuracy (89.4%–106.5% recovery).
• Achieved high sensitivity with limits of detection (LOD) ranging from 0.02 to 0.98 ng/mL.
• Successfully quantified specific gelatin peptides using the response-boosting protocol.

Conclusions:

• The developed response-boosting strategy significantly enhances detection sensitivity for gelatin quantification.
• This protocol offers a reliable and promising approach for the quality control of food products containing gelatin.