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Increasing the energy separation between ε1 and ε2 will decrease the population of ε2 in a Boltzmann distribution. The Boltzmann distribution describes the distribution of particles among energy levels at a given temperature. The equation is: Ni/Ntotal = exp(-εi/kT), where Ni is the number of particles in energy level εi, Ntotal is the total number of particles, k is Boltzmann's constant, and T is the temperature. As the energy separation between ε1 and ε2 increases, the exponential term for ε2 becomes smaller, resulting in a lower population in that energy level.