One way is to test samples for bioburden at the start and end of the hold time, and note any significant differences.

Dear Rishabh,

Attached please find the required guidelines.

Hoping this will be helpful,

Rafik

You may find required information on the link

http://www.pharmaguideline.com/2015/06/hold-time-study-in-pharmaceutical-manufacturing.html

Also you may find the attchment

WHO TRS 992 “Annex 4 General guidance on hold-time studies”

Is there any guidelines for in process hold time study of semi-solid dosage form