We report that the p-xylylenediammonium ion (11) acts as a template in the cucurbit[n]uril forming reaction that biases the reaction toward the production of methylene bridged glycoluril hexamer (6C) and bis-nor-seco-CB [10]. Hexamer 6C is readily available on the gram scale by a one step synthetic procedure that avoids chromatography. Hexamer 6C undergoes macrocylization. with (substituted) phthalaldehydes 12,14,.15, and 18-in 9 M H2SO4 or concd HCl at room temperature to deliver monofunctionalized CB[6] derivatives 13,16,17, and 19 that are poised for further functionalization reactions: The kinetics of the macrocyclization reaction between hexamer and formaldehyde or phthalaldehyde depends on the presence and identity of ammonium ions as templates p-Xylylenediammonium ion (11) Which. barely fits inside CB[6] sized cavities acts as a negative template which slows down transformation of 66 and paraformaldehyde into CB[6]. In contrast, 11 and hexanediammonium ion (20) act as a positive template that promotes the macrocyclization reaction between 6C and 12 to deliver (+/-)-21 as a key intermediate along the mechanistic pathway to CB[6] derivatives. Naphthalene-CB[6] derivative 19 which contains both fluorophore and ureidyl C=O metal-ion (e.g., Eu3+) binding sites forms the basis for a fluorescence turn-on assay for suitable ammonium ions (e.g., hexanediammonium ion and histamine).