Abstract: A pair of diastereomers 2 a and 2 b were synthesized from the conjugation of bridging chiral de‐tert‐butylcalix[4]arene racemate 1 and (1S)‐(+)‐10‐camphorsulfonyl chloride. The diastereomeric separation by crystallization was explored in different solvents, temperature and time. The optimal crystallization procedures can afford them in a desirable yield, de and purity. A pair of enantiomers 1 a and 1 b were subsequently obtained after their hydrolysis. Moreover, their absolute configurations were determined by X‐ray crystallographic analysis. The crystallization results can match the large‐scale preparation of optically pure de‐tert‐butylcalix[4]arene in laboratory and industry. [ABSTRACT FROM AUTHOR]