The objective of this study is to evaluate the effect of hydrogen on the micro- and macro-strain of austenitic stainless steel using X-ray diffraction. When hydrogen is trapped in lattice sites, it can affect both the micro- and macro-strain. The micro-strain was evaluated through fitting profiles to measured X-ray diffraction profile using a fundamental parameter method. The macro-strain, i.e., the residual stress, was evaluated by a 2D method using a two-dimensional PSPC. The experimental samples were charged with hydrogen by a cathodic charging method. The results revealed that the induced residual stress was equi-biaxial and compressive, and that the micro-strain increased. Both of these varied rapidly with increasing hydrogen charging time. Saturation occurred at a compressive stress of around 130 MPa. On reaching saturation, the hydrogen charging was terminated and desorption of hydrogen began at room temperature. Then, the strains decreased and the compressive stress reverted, ultimately, to a tensile stress of 180 MPa. Martensitic transformation occurred due to hydrogen charging and this had a significant effect on the X-ray diffraction profile.