Methods for separating inorganic compounds from organic materials in investigation of trace proteins in soil

Albert, J. D.

Walwick, E. R.

Zalite, B. R.

English

NASA Technical Reports Server

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. -  I Reporting Period: 1 Mar& 1965 - 3 1  May 1965 
Prepprod by: E. It. Uahrick 
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https://ntrs.nasa.gov/search.jsp?R=19660017747 2020-03-16T19:12:31+00:00Z
Research during th i s  mparting period has been on the extraction - -- and 
preparation of organic materials from soi1,and the interaction - of these materials 
with the thiacsrbocyanlne dye. =e approach has been to:  (e) determine the 
tendency of inorganic polyanionrr uhlch form ar are s table  i n  aqueous systems t o  
change the dye spect-, (b) &same tbe rac t ion  of the dye with soluble and 
col loidal  inorganic polpnions extractable frapp s o i l  a d  (e) develop a sequence 
of preparative stape n e c e a s a z  ts separate caapounds of classes a and b fran the  
polymeric organic m t e r h l a  extractable frosa soils whilemaintaining an optimum 
extraction of the organic arterial. 
--- 
Procedures befng studied for the extraction of so i l  organic material 
and the rationale for the i r  use were reported i n  the  previous quarterly report. 
The a a ~ l y s h  of carbon values war not cuuplete, but f.8 now available and the 
results are sham in Table 1. Q r i e f l y  the extraction procedure consisted of 
suspending the soi  1 in the pretrertaerrt solut ton for 1 bour with shaking. 
s o i l  was separated by centrifugstion, ~18hed f ive thes v i th  water a d  shaken 
i n  0.5 I loaQfI for one hour. 
c e n t r i f w t f o n ,  6 e  8upatarte MI exhaustively dialyzed spinst distilled water 
and the precipitate (extracted soil) was dried and s r l y s e d  for  carbon, Several 
interesting pointa are apparent. 
carbon, l h i s  is an ideal resul t  for a pretreatment, since it results in no loas 
of orgaaic material. Because of t h l s  favorable r e s u l t ,  the experhent w a s  
r e p a d ’  ?ad the previous resu l t  was confirmed. 
the lactate pretreatment plue the rubeequent sedirrm hydroxide extraction vas 
rwgbly t&e same 98 that r m e d  by the sodium hydrosLde extraction of the 
pl rqphormta  ptetreated ao i l ,  but there waa twice a0 lpeh non-d ia lpb le  tarboa 
reamed by the fsctats system. The EDTA extraction sy8tesm .(pretreatment + NaoH 
extraction) a l r a c t d  as much nondtolyzablc carbon as the pjraphoephate ayst-, 
but only 1 l i t t le  more than half as much t o t a l  carboa from the s o i l ,  
carbon rentwed by the various pretreatment. was not subjected to dialyeis so it 
is not apaarant hem mu& was n d i a l y z a b l c .  
removed by 0.1 M ECI must be fulvie acid slace it is soluble l o  acfd. 
tre!atments were carried out at acid pH and therefore anything removed by these 
solutioae would a l so  be considered fulvlc acid. 
The 
The so51 resfdue w a s  separated fran the extract by 
 
/ The pyrophosphate pretreatment reanoved no 
The organic carboa renwed by 
Ihc argunkc 
Hasever, by defini t ion t h e  carbon 
A l l  pre- 
_ _ -  
Ihe resu l t s  ehar that with the epception of pretreatment with water, 
each of the pretreatment ecbmee vaa effective in increasing 
nondialymble o r e i c  material extracted by 0.5H UOB. 
phosphate demorrstrrte that pretreatmint can ba wed which w i t 1  not reaaave 
orgpnic mwerm$ras the roil, but w i l l  aid in iacrg.sf=ag tbe asmunt of 
material oubsequcatly extracted with alkal i .  It MIS amticfpated that this  
would ba! the type of resul t  obtained with the RC1 pretreatmat, on the bash 
of the r e m ~ f t a  obtained by Qtadhrf srd Stevenson @roc, Soil Science Soc. 
of h r . *  2, 508-5U (1957) ). 
cated €IC1 pretreatment greatly increased (3 fold in  one case) the humic acid 
s o l u b i l i ~ e d ,  but did not greatly increme the total carbon extracted. 
camp8rleoa of our result8 with the i r s  point8 up the problems encountered in 
a t t empt fq  t o  devise a universal soil extraction procedure. 
the amount of 
The reeults with pyreL 
Their r m u l t e  for four clay type Soil8 indi- 
The 
%e main point t o  be made ra a result  of the foregoing work is the 
appropriateness of a pretrestnmnt prior to alkali extraction. 
now being a c t d e d  t o  iacfda rrlailnr studies with desert t p  roils. 
felt  that t t y  Containhg Soil8 Offer the greatest d i f f icu l t ie8  
extraction of organic saterklr a d  ab0 for interference wtth the dye 
reaction. 
reference for k t e r  work with deeart soils, 
Thir work is 
It vas 
in the 
It yds a l so  f e l t  that work v i th  the clay roi ls  would serve as a 
Adopting the premise that  a pretreatment w i l l  be included in  any 
r o i l  extraction scheme adopted greatly simplifies the problem of exclusion and 
removal of inorganic materials that will interfere with the dye reaction. 
Previow studies of the dye reaction with various inorganic materials were 
presented in the last quarterly report. Theee rtudies were concerned w i t h  the 
repoval of t h e  matariala by u l t r a f i l t r a t ion  a d  dialysis.  
r epu ted  with the incltuioa of an acid extraction and water trorh prior to 
0.5# #a08 extraction, 
remmal of such polyaaioas as mt&phospbate, isopo~y a d  beteropoly acids. 
The materials studfed were choeen kuuse of thefr interferance with the dye 
reaction fn auy of rreveral ways, formation of p o l p e r k  speciea in  aqueous 
solut Lon, suspension as colloids, diff icul ty  in dialysis,  greater solubi l i ty  
B e r e  studies were 
The &vantage gained by the acid and water wash is the 
-2- 
in alkali and lower so lubi l i ty  at xmutt.1 pE. 
elbiaatad f r aa  concern because of the i r  so lubi l i ty  a t  a concentratton of 
2 gm per 10 a1 of 0.U Ilcl were potasriur pyroeulfite, potarei\lsr pyrosulfate 
sodium arPhate, aodilaa pyrophmphrte, eodimmetapho6pbate, sodium ortho 
phoephate, p8orpSrotungrtic acid .nd 8od%um m o s u l f i t c .  In caaes it was 
neca8uary te add cencentrated acid t o  warccme the buffering of the t ea t  
PateFhls 8pd obtain a pE of at hast 1.0 during the acid pretreatment. 
the  ortezirb 8tudied only a h  (AlK(S04),) was not soluble in acid, but waa 
dissolved in the sub8equent water wash. Materials not soluble I n  the acid 
srtraetioa QY aubseqtreat water wsh were subjected t o  a l lu l tne  extraction. 
The a l k d h e  =tract v.8 divided rad ollt aliquot wacl dialysed againnt vater 
and the ether agrhst 0 . M  alkali a d  then d ie t i l l ed  water. 
potassium psreulfate, sodium aroenfte, f e r r i c  oxide, poly oulf ide, aluminum 
bydrarride and plumbow hydraride pve no dye r d c t i o o  a f t e r  e i ther  water or 
a l k a l i  and water dialyeis.  Ebttractr f r o m ~ l i n ,  bentonite, uralite, sodium 
silicate @azO:S102, 1:2.86Ia poturrftm silicate ($0:siO2, 1:2.10) and send 
cawed dmqp In  dye apeecna a f t e r  d w t i v e  water d i a l y s b ,  But not 
a f t e r  alkali dk1yai.s follawrd by water dialysfs.  
aluminosilicate, of? the four tested(iuoUn, bentonite and ura l i te )  which s t i l l  
&.ve a pomtive te r t  a f t e r  the caoplete treatment. Because of the ubiquity of 
alumfnc?8ilicatas Ln soife, it was f e l t  that  these experiments should be repeated 
several  t f n e s  t o  establish the reproducibility of the resul ts .  
done and the resu l t s  were found t o  vaty with time of dialysis .  
r i l i c a t a s  could be removed with -tar dialysis  i f  the dialyslr; period was 
auff ic ient ly  long. 
prior t o  the rUralinc ut rac t fc re  appears t o  be required t o  e l k i a a t e  them by 
dialysf. .  
Teat materials wbich could be 
Of 
* 
Extracts f r m  
Phlogopite was the only 
Thfs has been 
The soluble 
'In the  case of a a ~ e  of the aluminosilicates acid extraction 
inergaa%c o r t e t t b  cheseu for  t h i s  study const i tute  a broad 
spectrum of inorgadc chemical typae which could of fe r  d i f f i c u l t i e s  t o  the dye 
detection ryotan, 
can be reaKnred by a process of extraction vhich would be required in any event 
1Eo f ree  .oil organic material f raa  inorganic constituents. 
ublquitp of the aluminosi l~cates  in surface materials, it  is f e l t  further 
studier -1d be carried out concerning the i r  removal from thecre s o i l  extracts.  
Hawever ,  t h k  prasent work hat ahawn that these matarfals 
Because of the 
In the care of the phlogopite preparation it is necessary t o  determine why 
it fr not as earrfly elhinated a. are the other aluarinosilicates studied. 
The pilsitive dye reaction obtained w i t h  the alkaline dials+& phwgapite 
c.0 not be 8 resul t  of reaction w i t b  extracted orgadc carbon. 
mrbaa r a r r l p i r  aa a l l  four of the alurinosil icatee did not iadicate P 
eraee of c.rb#a Gr t8ese n h o r i s .  
Organic 
It -it be noted that the organic material extracted fran the 
rtaodard clay 60f1 gives the 8 8 ~  dye spectruPa a f t e r  a b l f s e  dialysi8 81 
with the water dialysis  aloaa. 
alumfnosilicate extracts. 
This is in contrart t o  the resul ts  with the 
Large scale extract iars  of the clay r o i l  (used ae a rtaPdard) 
and desert r o i t s  w i l l  be carried out with the praporred axtraceion procedure 
(ire.,  ectd pre-artraetioa, water waah a d  alkali extraction). 
extraction product w i l l  be subjected t o  infrared aaalyeis t o  determfne the 
relat ive coucentration of functfonal group8 present In these araterhls .  'Ihe 
elimiuatian of s i l ica tee  fran there products w i l l  be eetablished. Selective 
rewrval of the carboxyl g ~ 3 i i p s  will be carried out. 
the decarboxylated product a d  w l t h  the untreated ma te r id  will be cappared. 
(pa eet8blfsh the nature of the dye reaction. Par t fa l  axidation a d  reduction 
of the soil extract aoteri.1 w i l l  beatcrpted t o  determine its effects on the 
dye reactfoa. 
Tbe dfalyzsd 
k c t i o n  of the dye v i th  
StuiHea w i l l  be carried out Ln vhich the alminomilfcater, kaolin, 
bentonite, uralite and p-opfte v i l l b e  subjected t o  the extraction procedure. 
The alkaline extract w i l l  be dlalysed for various periods agaimt alkali to 
determine the t h e  required t o  runwe the alunlnollilfcates. 
membrane fllterr w i l l  be r q l q e d  wi th  the phlogopite extract t o  determine i f  
thfa w i l l  improve its subrcqmut d i a l y s h  rate.  
Smaller pore 
4- / 
Retreat ing -
blanc 
RZ0 
.lx ficl 
--- 
lone 
21 
15 
9 
Z Carboa removed 
by 0.SN W.Oa 
(+ 88 > 
18 
34 
l5 
40 
20 
54 
X Na.t-dia&able 
carboa removed 
by NaOff 
2.96 
1.63 
3 , s  
10.4 
4.90 
4.80 
Sail  caplOpea in these studies waa a local agrlcultwal clay cantaining soil. 
Tbe organic carbon content of t h i s  roll ie 0.85 percent. 
* EthylenedtaPipatetrWcLtir r c i d  


Experimental Studies for the Detection of Protein in Trace Amounts Quarterly Status Report, 1 Mar. - 31 May 1965

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