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1.They did not add these extra hours and minutes and seconds.

2.They did not add these extra hours, minutes, and seconds.

Are there any difference in these two sentences when using "and" instead of comma in a list?

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    The sentence with the comma is better, as when speaking you would not say "and" repeatedly. – Burhan Khalid Apr 9 '17 at 8:33
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Commas Rules
Use commas to separate words and word groups in a simple series of three or more items. Like in your sentence:
They did not add these extra hours, minutes, and seconds.
This is the correct way to write it.

Another example like:
We had coffee, cheese and crackers and grapes.
In this case we have to use Oxford Comma (when the last comma in a series comes before and or or)
We had coffee, cheese and crackers, and grapes.
Adding a comma after crackers make it clear that cheese and crackers represents one dish.
Conclusion
Use comma to separate adjectives, nouns, verbs, words, word groups and use and just before the last item unless it is needed.
Example:
To make her famous muffins, Paulette bought fresh broccoli, baker's chocolate, flour, sugar, and eggs.

References:
The item in a Series-Grammar Bytes
GrammarBook.com

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