Aerogen Ultra (AU), a unique holding chamber designed to be utilized with the Aerogen Solo vibrating mesh nebulizer (AS), was developed to maximize the total dose that subjects can breathe in. It can be used for continuous and intermittent nebulization with the elective supply of additional oxygen. This work's aim was to see how well an AS coupled to an AU as a holding chamber with a mouthpiece and valved facemask (VFM) work at diverse oxygen flows from 0 to 6L/min. Methods: this was an In-vitro and in-vivo study, First, in the in-vitro part, with 500 mL tidal volume, fifteen breaths every minute, and a ratio of 1:1 between inhalation and exhalation, we were able to imitate adults' natural breathing patterns. Five evaluations were done for the combination of AS and AU adaptor with VFM or mouthpiece and 0, 2, 3, 4, and 6L/min oxygen flow. Secondly, we conducted an in-vivo trial with 12 healthy non-smokers who were > 18 years old and had a typical forced expiratory volume in one second (FEV1) that was ≥ 90% of predicted. The subjects used AS linked to AU with VFM or mouthpiece and 0, 2, 3, 4, and 6L/min oxygen flow to inhale Salbutamol nebulized in 1 mL (5,000 µg) during normal tidal breathing. Urine was taken 30 minutes after dosage as a measure of lung deposition, and urine was collectively gathered over the course of 24 hours as a measure of systemic absorption. Results: The salbutamol amount deposited on the inhalation filter and the amounts of excreted salbutamol in the 30 minutes and over 24 hours after the start of the inhalation were improved until oxygen flow of 2 L/min, and 3 L/min with the mouthpiece and VFM, respectively. In comparison to the other oxygen flows, this flow had significantly greater salbutamol and the delivered dose was then steadily declined until at 6L/min of oxygen flow, p < 0.05. Conclusions: With the AS connected to the AU, mouthpiece, or VFM, the total inhalable doses and the amount of excreted salbutamol within the first 30 minutes and over the course of 24 hours after the start of inhalation were inconsistent at different oxygen flows. Up until oxygen flow reached 2 L/min with the mouthpiece and 3 L/min with the VFM, there were notable improvements; however, beyond that, there was a steady decline to lower values at 6 L/min of oxygen flow.