When I make Chitosan microsphere(CM), I use 1% of Chitosan in 0.5% acetic acid.

After adding TPP, I harvest CM and then wash it with DI water just one time.

(At this moment, It form pellet very well)

But, when i try to do second washing, it never form pellet..(13000g & 10min centrifugation.)

I don't know what is happening.

Anybody know this, please tell me, please..

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