API nitrate test kit:
Bottle#1 is primarily HCL (Hydrochloric Acid and other proprietary ingrediants)
Bottle#2 is polyethylene glycol polyethylene glycol - this a reagent water soluble dye
The HCL exchanges nitrate ions for chloride ions. Bottle#2 reagent then reacts with chloride ions and provides a color for results.
This what i remember from chemistry (adding H/CL and
NO3 together/ CL HNO3) and reading the MSDS of API Nitrate Test. So if you don't agitate the tube on step 1, it can/will affect the final result.