13S 
cstiniaLiuu  (if  liotli  uuallcrci^l  hacmof^icjhiu  and  alsu  Inicmoglubini 
whicli  had  fieen  broken  up  by  the  action  of  the  urine. 
MeJhod  I.  The  urine  was  centrifugalised  and  the  supernalaiii| 
liquid  diluted  with  a  measured  volume  of  distilled  water  until  tliel 
percentage  of  haemoglobin  as  judged  by  the  naked  eye  was  aboiit| 
O’ 1 5  per  cent.*  The  diluted  urine  was  then  put  into  the  half  celln 
von  Fleischl’s  haemoglobinomcter.  If  the  dilution  was  found  unsuil 
able  a.  fresh  dilution  was  made  until  a  concentration  was  olitaiiidj 
which  could  be  ea.sily  matched.  The  haemoglolainometcr  scale  wasj 
previously  standardised  by  means  of  solutions  containing  knoivii| 
percentages  by  volume  of  healthy  human  red  cells  ;  this  having  bcci| 
done,  the  percentage  of  haemoglobin  in  the  diluted  urine  could  beal| 
once  determined  from  the  scale  numbers. 
In  many  cases  no  difficulty  was  found  in  making  a  dcterminai«| 
of  haemoglobin  by  this  method.  Difficulty  arose,  however,  when  tlit| 
haemoglobin  had  been  largely  converted  intrj  the  brown  substaiio| 
already  referred  to,  a  change  which  is  likely  to  occur  when  the  iiri« 
has  been  retained  in  the  bladder  for  a  long  time  before  being  passei 
or  after  being  voi(.led,  has  been  allowed  to  stand  for  some  tune  befoti 
examination.  This  change  is  of  course  relatively  more  marked  11 
urines  containing  small,  than  in  urines  containing  large,  percentagn 
of  haemoglobin.  Another  source  of  difficulty  in  matching  the  colon 
of  the  diluted  urine  with  that  of  the  lintecl  glass  of  the  haeraoglobnii: 
meter  occurs  when  dissolved  haemoglobin  is  discharged  in  5iiij!| 
amount  into  urine  which  is  itself  of  a  dark  brownish  amber  co!oiii| 
as  not  iinfrequently  happens  in  malarial  patients.  Matching  alt; 
becomes  clifRcult  when  the  urine  in  the  bladder  contains  bacteiia,)! 
occurred  in  Case  2,  the  turbidity  thus  caused  not  being  completel'* 
removable  by  centrifugalisation.  When  from  any  of  these  causes' 
becomes  impossible  to  employ  the  above  described  method  for  tk 
estimation  of  dissolved  haemoglobin,  recourse  may  be  had  to  d. 
next  method. 
Method  2.  The  urine  was  diluted,  if  necessary,  with  a  kiio«. 
volume  of  distilled  water  until,  in  a  column  one  to  two  centiinetifi 
=!=  This  rough  comparison  may  be  made  by  pulling  llie  cenlrifugalised  urine  inloiitq 
lube,  lining  anolher  le.sl-lube  with  a  0-15  per  ccnl.  solulion  of  haemoglobin  and  comp 
Ihe  Iwo,  Ihe  Ibtraer  being  rurlher  diluled  or  a  lesser  dilulion  prepared,  as  may  be  necess^ 
Wilh  a  Hllle  praclice,  however,  il  becomes  jrossible  lo  obtain  the  retiuired  dilution'" 
suflicient  upproximaLioii  by  ihe  naked  eye  without  comparison. 
