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Nitrous oxide (N2O) is an important greenhouse gas and is the third-largest contributor to global warming with a global warming potential (GWP) higher than that of carbon dioxide. Ambient N2O is monitored globally and described at nanomole per mole (nmolmol(-1)) levels. Metrologically traceable and highly accurate N2O gas standards are required to understand the behaviour of the greenhouse gases in the atmosphere and GWP. A group of primary standard gas mixtures (PSGMs) in aluminium cylinder material has been prepared in synthetic air for the calibration of analytical instruments for measurement of N2O at an ambient level. An automatic weighing system with a mass comparator and a fully automated circular rotary plate, as well as a gas filling station, was used in the preparation of the gas mixtures. Analytical methods such as gas chromatography coupled with micro-electron capture detector and cavity ring-down spectroscopy were adopted in the verification process. Four PSGMs in the final preparation step were used to analyse unknown background and filtered air samples at the ambient level. An expanded uncertainty relative to the gravimetric amount fraction was found to be 0.02%. Results show that the background air samples lie within the calibration range with 329 to 330nmolmol(-1) obtained and 135nmolmol(-1) obtained for filtered air sample.