We calibrate the X-ray measured hydrostatic equilibrium (HE.) mass and assess the origin of the H.E. mass systematics using 2-D spectrally measured X-ray properties. We obtained that the average X-ray mass derived from H.E. using XMM-Newton data is lower compared to the weak lensing mass from Subaru data for relaxed clusters in a sample of 12 clusters at z similar to 0.2. This is comparable to the expectation of numerical simulations because of the non-thermal pressure support due to turbulence and bulk motions. The gas mass to weak lensing mass ratio shows no dependence on the cluster morphology, which indicates that the gas mass may be a good mass proxy regardless of the cluster dynamical state. To understand the origin of the systematics of the H.E. mass, we investigated 4 nearby clusters, for which the substructure is quantified by the radial fluctuations in the spectrally measured 2-D maps by a cumulative/differential scatter profile relative to the mean profile within/at a given radius. The amplitude of and the discontinuity in the scatter complements 2-D substructure diagnostics, e.g. indicating the most disturbed radial range. There is a tantalizing link between the substructure identified using the scatter of the entropy and pressure fluctuations and the deviation of the H.E. mass relative to the expected mass based on the representative scaling relation, e.g., M-M(gas), particularly at r(500) - the radius within which the over-density, Delta, is 500 with respect to the critical density. This indicates that at larger radii, the systematic error of the H.E. mass may well be caused by substructure.