Plans for six-day, 24 hour opening of the Tesco superstore in Princes Road, Chelmsford, were turned down by the borough council's development control sub-committee on Monday.
Councillors rejected advice from their officers that they should allow a one year trial period of the round-the-clock opening, (excluding Sundays) in order to assess the impact on neighbours.
The store is at present open from 8.30am- 9pm Monday to Friday, 8am-9pm on Saturdays, and 10am-4pm on Sundays.
The sub-committee was told that Tesco's wanted to provide a greater choice of shopping hours at Princes Road following customer demand and successful 24-hour trading elsewhere in the country.
Although the store's noise consultants had reported that trading could be extended round the clock without adversely affecting the amenities of local residents, there had been a number of objections from neighbours and from the Miami Hotel.
Residents mainly of Bruce Grove and Hillside Grove had objected on the grounds of potential noise and disturbance, and the hotel had claimed that associated vehicle and headlight movements would create unreasonable disturbance for its guests.
But council officers pointed out there had been only a low number of complaints received by the authority in respect of the existing use, and added that there was an assumption that stores are now able to trade 24 hours a day in similar areas without detriment to local amenity.
Cllr Mrs Freda Mountain, whose ward includes the site, said she was delighted with the decision.
"There is no need for the store to be open 24 hours in this day and age," she said.
Essentials such as bread and milk were already available round the clock at the garage on site, and, she said, she understood the recently-introduced Internet shopping was very popular.
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