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I tried using the following methods:

Using a unlocker (to ensure that no files are being locked)

Using the method Process Explorer (to find if there are any files currently being used)

but I still could not be able to remove my USB flash drive from my Windows.

Am I missing anything that prevent me from safety remove my USB flash drive? (such as Antivirus running background but scanning the files on the thumbdrive?)

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Sometimes Windows just doesn't want to let go. You can just go ahead and pull the drive and hope for the best -- generally this won't harm anything if no writes are ongoing at the time. Or you can shut down your box. And I'm pretty sure it's sufficient to hibernate your box. – Daniel R Hicks Jan 16 '12 at 4:18
This may be a the same… it may be do to Windows indexing. – N4TKD Jan 16 '12 at 4:18
@DanH, while you may be right, I wouldn't want to test the hibernate thing with an important thumb drive. That's me being seriously overcautious, but still, powering down is the very safest way to remove a drive that Windows won't release. – user3463 Jan 16 '12 at 4:26
Does logging off the user account then log back in work? – Moab Jan 16 '12 at 5:45
Could it be the antivirus scanning the files on the thumbdrive? – Jack Jan 27 '12 at 7:38
up vote 0 down vote accepted

USB drives that don't eject properly are one of my biggest hobbies! :P

In all seriousness though, one thing that often works for me if a drive just doesn't want to be "removed safely", is to eject it through Windows Explorer.
Ejection feature fails

When viewing the "Computer" section where it lists all drives, you can right-click the device and select "Eject".
Usually this performs the same action (or so it seems) as the "Remove hardware safely" option. But it can go an extra step. If initial unmounting failed, it might ask you if you want to force ejection of the device. Sometimes this does work for me. Explorer going an extra step

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