Introduction: Stabilizing adjuvants plays an important role in vaccine formulation. They make it possible for some preparations to resist longer to higher temperatures without losing efficacy. Lactose is a disaccharide composed of galactose and glucose condensed by β-1→4 glycosidic linkage. It works as a part of formulation as a stabilizing agent in a wide scope of vaccines produced by Bio-Manguinhos/ FIOCRUZ such as HiB (Haemophilus influenza B), Meningococcal AC and MMR (measles, mumps, rubella). It is important to control lactose concentration in formulations for quality control purposes. In this context, High-Performance Anion- Exchange chromatography with Pulsed Amperometric Detection (HPAE-PAD) is a powerful technique to determine carbohydrates in pharmaceutical products, offering advantages such as accuracy and selectivity with minimal sample preparation, cost-effectiveness, and rapid analysis compared to other analytical techniques. Objectives: Develop an analytical method using HPAE-PAD for determination of lactose in 15,75% (w/v) stabilizer solution for vaccine production. Methodology: This study used an Ion Chromatograph Professional IC Vario 940 Methrom, with a Pulse Amperometric Detector, a CarboPacTM PA10 column, and a BorateTrapTM guard column. The mobile phase was sodium hydroxide 100 mM. A five-point standard curve was prepared, phase overing a concentration range from 0.5 to 10.0 ppm. Subsequently, the method was tested for its linearity, precision, and accuracy, by RDC166/17. Lactose USP standard and six independently prepared solutions containing 15.75% (w/v) lactose as a stabilizer were employed to check method performance. The following acceptance criteria were applied: R2 value greater than or equal to 0.99, recovery rates within 80% and 120%, and a Relative Standard Deviation (RSD%) below 10%. Results: The method demonstrated linearity with a correlation coefficient value of 0.9998. The triplicate preparations of the calibration curve exhibited RSD% values ranging from 3.3 to 8.79%. Analyst 1 and 2 provided RSD% values, in precision study, of 7.2 and 3.0, respectively, while the intermediate precision showed a 6.5% of RSD%. The recovery values for the six individual determinations ranged from 98 to 116%. Conclusion: The proposed method for quantitation of lactose in 15,75% (w/v) stabilizer solution by HPAE-PAD shows good precision, linearity, and accuracy and it is suitable for analytical validation. Due to its applicability to Lactose, the HPAE-PAD technique could be used for the determination of otherHPAE-PAD saccharides with importance for vaccine development and production.