Spirituality and spiritual well-being are connected with many areas of human life. Thus, especially in secular countries, there is a need for reliable validated instruments for measuring spirituality. The Spiritual Well-Being Scale is among the world’s most often used tools; therefore, the aim of this study was its psychometrical evaluation in the secular environment of the Czech Republic on a nationally representative sample (n = 1797, mean age: 45.9 ± 17.67; 48.6% men). A non-parametric comparison of different sociodemographic groups showed a higher disposition for experiencing spirituality among women, older people, and divorced persons. Based on confirmatory factor analysis, negatively worded items were excluded using a polychoric correlation matrix. The new version of the scale consisting of 11 items had good internal consistency (Cronbach’s α = 0.85; McDonald’s ωt = 0.91). The two-factor model of this shortened version, with factors corresponding to the Religious and the Existential subscales of the Spiritual Well-Being Scale, shows a satisfactory fit with the data, where the loadings of all items ranged from medium to high. Thus, this study offered a new version of the tool, convenient for measuring spiritual well-being in secular conditions.