Purpose: The skin acts as a barrier to prevent external harmful substances from entering the body. This function interferes with active ingredient absorption. Understanding the degree of absorption of the substance into the skin is important to predict the effectiveness of cosmetic active ingredients. This study aimed to investigate the human efficacy of Åbsoro-Pep™ substance, which contains gold cysteinyl hydroxy-prolyl dipeptide-48 as a key ingredient, for improving absorption. Methods: This study conducted a human application test to evaluate the improvement in skin absorption (including absorption rate, speed, and depth) of Åbsoro-Pep™ using three-dimensional Raman spectroscopy, targeting adult women aged 20–68 years. Results: The results revealed that skin absorption rate, speed, and depth were all significantly enhanced (<i>p</i><0.001) 30 min after application of the test substance compared to before application. Additionally, a high-concentration test confirmed that substance A application site was more helpful than substance B application site by improving skin absorption rate, speed, and depth by 1.80, 2.13, and 2.13 times, respectively. Conclusion: Åbsoro-Pep™, which combines gold nanoparticles with peptides, is expected to be predominantly used and applied in functional cosmetics to improve skin absorption.