Supporting

 Information  
 
Cleaning  with  Bulk  Nanobubbles  
 

Jie   Zhu†,   Hongjie   An‡,   Muidh   Alshebri‡,   Lvdan   Liu†,   Paul   M.   J.   Terpstra#,  

Guangming  Liu*,†,  and  Vincent  S.  J.  Craig*,‡  

 

†Department   of   Chemical   Physics,   Hefei   National   Laboratory   for   Physical  

Sciences  at  the  Microscale,  University  of  Science  and  Technology  of  China,  Hefei,  

P.R.  China  230026  

‡Department   of   Applied   Mathematics,   Research   School   of   Physical   Sciences   and  

Engineering,  Australian  National  University,  Canberra  ACT  2601,  Australia  

#  Consumer  Technology  Research  Institute,  Wageningen,  Netherlands    

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

S1  
 
Table   S1.  Size  and  concentration  of  nanobubbles  in  ec-­‐H2O  sub  450  nm  over  5  
days.  

Day   Mean  (nm)   Mode  (nm)   Concentration  

(  particles  x  106  /mL)  

1st  day     96  ±  2   83  ±  4   483  

2nd  day   110  ±  2   91  ±  5   432  

3rd  day   120  ±  4   99  ±  5   161  

4th  day   116  ±  2   94  ±  10   121  

5th  day   97  ±  4   91±  11   169  

   

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

S2  
 
Figure  S1.  Adsorption  inhibition  study  performed  using  ellipsometry  to  measure  
the   thickness   of   adsorbed   BSA   to   a   silicon   wafer   as   a   function   of   time.   (a)  
Comparison   of   BSA   adsorption   from   NaCl   solution   (2.1   mM)   with   adsorption  
from   ec-­‐H2O   sub   20   nm.   (b)   Comparison   of   BSA   adsorption   from   ec-­‐H2O   with  
ec-­‐H2O   sub   450   nm.   The   comparisons   demonstrate   that   nanobubbles   are  
responsible  for  the  reduced  adsorption  of  BSA  in  the  presence  of  ec-­‐H2O.  
 
 
 
 
 
 
 

S3  
 
  
Figure  S2.  Adsorption  inhibition  study  performed  using  ellipsometry  to  measure  
the   thickness   of   adsorbed   lysozyme   to   a   silicon   wafer   as   a   function   of   time.   (a)  
Comparison   of   lysozyme   adsorption   from   NaCl   solution   (2.1   mM)   with  
adsorption  from  ec-­‐H2O  sub  20  nm.  (b)  Comparison  of  lysozyme  adsorption  from  
ec-­‐H2O   with   ec-­‐H2O   sub   450   nm.   The   comparisons   demonstrate   that  
nanobubbles   are   responsible   for   the   reduced   adsorption   of   lysozyme   in   the  
presence  of  ec-­‐H2O.  
 
 
 
 
 
 
 
 
 
S4  
 
Figure  S3.  Adsorption  inhibition  study  performed  using  ellipsometry  to  measure  
the  thickness  of  adsorbed  BSA  to  a  hydrophobised  silicon  wafer  as  a  function  of  
time.   (a)   Comparison   of   BSA   adsorption   from   NaCl   solution   (2.1   mM)   with  
adsorption   from   ec-­‐H2O   sub   20   nm.   (b)   Comparison   of   BSA   adsorption   from  
ec-­‐H2O   with   ec-­‐H2O   sub   450   nm.   The   comparisons   indicate   that   the   nanobubbles  
are   preventing   fouling   of   the   surface   and   the   chemical   changes   associated   with  
the  electrolysis  of  water  may  also  contribute  to  the  fouling  prevention.  
 
 
 
 
 
 
 
 

S5  
 
Figure  S4.  Adsorption  inhibition  study  performed  using  ellipsometry  to  measure  
the   thickness   of   adsorbed   lysozyme   to   a   hydrophobised   silicon   wafer   as   a  
function  of  time.  Here,  all  the  thickness  of  adsorbed  protein  was  measured  in  air.  
(a)   Lysozyme   adsorption   from   NaCl   solution   (2.1   mM).   (b)   Lysozyme   adsorption  
from  ec-­‐H2O  solution.  (c)  Lysozyme  adsorption  from  ec-­‐H2O  sub  20  nm  solution.  
(d)  Lysozyme  adsorption  from  ec-­‐H2O  sub  450  nm  solution.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

S6  
 
  
Figure  S5.  Adsorption  inhibition  study  performed  using  ellipsometry  to  measure  
the   thickness   of   adsorbed   lysozyme   to   a   hydrophobised   silicon   wafer   as   a  
function   of   time.   (a)   Comparison   of   lysozyme   adsorption   from   NaCl   solution   (2.1  
mM)   with   adsorption   from   ec-­‐H2O   sub   20   nm.   (b)   Comparison   of   lysozyme  
adsorption  from  ec-­‐H2O  with  ec-­‐H2O  sub  450  nm.  The  comparisons  indicate  that  
the  nanobubbles  are  preventing  fouling  of  the  surface  and  the  chemical  changes  
associated   with   the   electrolysis   of   water   may   also   contribute   to   the   fouling  
prevention.  
 
 
 
 

S7  
 
 100%  
Percentage  of  Protein  adsrobed  to  
90%  
80%  
70%  
nanobubbles  

60%  
50%  
40%  
30%  
20%  
10%  
0%  
1.0E-­‐06   1.0E-­‐04   1.0E-­‐02   1.0E+00  
Protein  concentra2on  in  mg/ml  
 
 
 
Figure   S6   Calculated   protein   adsorption   capacity   of   ec-­‐H2O.   The   data   from   the  
histogram  presented  in  Figure  2  for  ec-­‐H2O  sub  450  nm  was  used  to  calculate  the  
surface   area   of   the   nanobubbles   per   mL   of   solution   by   multiplying   the   measured  
number  of  nanobubbles  in  each  bin  by  the  area  of  a  nanobubble  corresponding  
to  the  bin  size  and  summing  across  all  bins.  The  total  surface  area  calculated  was  
3.28  x  10-­‐5  m2/mL  of  solution.  Assuming  that  the  maximum  protein  coverage  is  2  
mg/m2,   which   is   typical   for   a   protein   on   a   solid   substrate,   the   calculated  
maximum  mass  of  protein  adsorbed  to  nanobubbles  per  mL  of  solution  is  6.56  x  
10-­‐5  mg/mL  of  solution.  This  value  is  subtracted  from  the  bulk  concentration  to  
determine   the   percentage   of   protein   present   in   solution   that   is   expected   to  
adsorb  to  the  nanobubbles.  
 
 
 
 
 

S8