text: it is hypothesized that magnetic compounds , in particle form , tend to have significant surface polarity . that net surface polarity is minimized by agglomeration of crystals of such materials . when these crystals are subdivided or disrupted , they tend to be unstable . in accordance with the present invention , the nascent ( and probably charged ) surfaces of these sub - particles are stabilized by the coating material which is simultaneously deposited on these surfaces as the parent particles are sub - divided . for that purpose , the coating material may be chosen with reference to its tendency to respond to the surface polarity of the deagglomerated magnetic particle and various coating materials will thus react differently with different magnetic particle materials . if the treating or disrupting technique is , or includes , ph modification , the effect of ph modification on sub - particle surface polarity and coating material polarity may also be a consideration . the coating material is selected in each case with regard to its ability to adhere to or to be adsorbed on the surface of the deagglomerated or sub - divided particle so that the stability of the reduced particle size product is retained , to form a stable suspension thereof . this invention provides significant advantages over molday and owen et al . for preparing colloidal materials beyond the inherent simplicity of the method . for example , if it is desired to attach compounds to metal oxide particles where the compound has reactive groups or activated groups for subsequently attaching other materials ( such as antibodies or enzymes ), the molday and owen et al . procedures are limited by the choice of such compounds . this is because these processes necessarily start by mixing such compounds with metal chlorides which themselves can react with the compounds and which also create acidic ph . further , base addition , usually ammonium hydroxide , is required to form the metal oxides and that can clearly have negative effects on a variety of activated chemical groups used for the subsequent coupling . by the direct adsorption methods of this invention , no such limitation exists . further more , the coating of this invention can be done under non - aqueous conditions or where water is a minor component of the system . in some instances , coating materials can have limited aqueous solubility . certain types of coatings , such as the long chain hydrocarbons suggested in the papell patent , have a detergent effect . these coatings stabilize the subdivided deagglomerated magnetic particles and produce a stable suspension , such as that taught in papell . in accordance with the present invention , these suspensions are produced , from magnetite and with sonication as the disrupting means , several orders of magnitude faster than in the ball milling process taught in papell . to produce resuspendable products , it is important to select a coating material which not only stabilizes the sub - divided magnetic particles , but does so with a coating which remains intact when the coated particles are removed from suspension , which is not the case with papell . magnetic compounds which may be used as the starting material in the present invention include the transition metal oxides , sulfides , silicides and carbides , optionally having different transition metals in a single magnetic compound , such as gd 3 fe 5 o 12 . preferred are the class of magnetic oxides known as ferrites , generally represented as mo . fe 2 o 3 in which m is zn , gd , v , fe , ni , cu , co , mg , in particular magnetite ( feo . fe 2 o 3 ). in addition to the transition elements taught by owen et al . and the ferrites , above , a class of magnetic metal oxide which does not contain iron can be coated as described in this invention . these compounds include oxides of combinations of 2 or more of the following metal ions : al (+ 3 ), ti (+ 4 ), v (+ 3 ), mb (+ 2 ), co (+ 2 ), ni (+ 2 ), mo (+ 5 ), pd (+ 3 ), ag (+ 1 ), cd (+ 2 ), gd (+ 3 ), tb (+ 3 ), dy (+ 3 ), er (+ 3 ), tm (+ 3 ) and hg (+ 1 ). they differ from ferrites in both appearance and magnetic susceptibility . the non - ferrites can take any color from white or yellow to green and even brown . this makes them particularly useful in spectrophotometric applications . non - ferrites are generally less strongly magnetic than ferrites and , as such , pass through hgms filters in magnetic fields capable of collecting ferrite based materials which permits selective magnetic retrieval . the non - ferrous oxides can be employed in place of the metal oxides described by whitehead et al . to produce silane coated magnetic particles which have the desirable properties given above . similarly , when the chlorides ( or sulfates ) of such combinations are employed with the methods taught by molday or by owen et al ., coated product having very desirable magnetic and spectral properties can be obtained . as indicated above , coating materials which may be used are preferably in aqueous suspension or solution , although suitable coating materials in non - aqueous solvents or as melts may also be used . the coating material is usually a synthetic or natural polymer and may be a protein or a nucleic acid . these materials are combined in a liquid mixture , usually including a third component such as water , to form a suspension . the relative proportions of these materials in this mixture is not critical . however , in general , the proportion of magnetic particles to coating material is from 1000 : 1 to 1 : 10 ( by weight ). to make stable suspensions of coated sub - particles of the magnetic material , the mixture may be treated in a number of ways to disrupt or sub - divide the magnetic particulate starting material . these include mechanical and chemical means , such as heat , vibration , irradiation , sonication , ph modification , or a combination of these . of these , sonication is much preferred . we have also discovered that the employment of high gradient magnetic separation ( hgms ) in the production of such particles gives a dimension to their production which heretofore has not been achieved or recognized . the molday and owen et al . processes utilize either centrifugation , gel filtration or &# 34 ; salting out &# 34 ; for manipulative procedures during the processing of colloidal materials . in the example herein , hgms has been used to separate colloidal particles from unbound coating substance . in processes where it might be desirable to chemically couple substances to such particles , hgms offers process facilitation which is readily scaled and highly efficient . hgms can also be used in concert with tuned magnetic fields to fractionate preparations based on their magnetic susceptibility / particle volume ratios . this permits convenient fractionation and production of particle preparations of discrete sizes . when used as nmr contrast agents , the size of the particle plays an important role in the metabolic fate of the material . hgms , again with tuned magnetic fields , can also be used to selectively capture particles where the magnetically responsive core contains transition element oxides having higher magnetic susceptibilities than others in the mixture . this concept could have utility in a system whereby colloidal particles having different magnetic susceptibilities as well as different bioreceptors could be mixed with a sample and removed by sequential hgms using increasing gradient field strength . not only can hgms be used to remove unadsorbed coating or unreacted substances and by - products from colloidal particles , it can also be utilized for immobilizing coated magnetic products and doing reactions on the immobilized material . thus , if it is desired to chemically modify a given coating , reactants can be added to magnetically immobilized material , the reaction performed in that state and excess reactant or other reaction products easily washed away . this concept , somewhat like peptide synthesis done on solid supports , lends itself nicely to sequential reactions . the following examples will serve to illustrate the principles of this invention ; but the range of these examples should not be construed as limiting the scope of this invention . all reagents and chemicals used in these experiments were of analytical grade and obtained from fisher scientific ( valley forge , pa .) unless otherwise specified . magnetite was prepared by mixing solutions of 3 . 0 and 1 . 5 mg / ml of ferric chloride hexahydrate and ferrous chloride tetrahydrate , respectively , with stirring at room temperature while raising the ph to 8 . 6 with nh 4 oh . the resultant magnetite was collected magnetically , washed 3 times with distilled water and resuspended in distilled water . the preparation so made contained 1 . 5 mg / ml magnetite . even after sonication ( fisher -- sonic dismembrator model 300 ) for 3 minutes at 70 % output , these preparations will not remain suspended for more than about 2 minutes . to coat magnetite particles by the method of this invention 0 . 5 ml aliquots of various concentrations of various coating materials were mixed with 0 . 5 ml aliquots of the 1 . 5 mg / ml magnetite suspension . samples were mixed in conical plastic centrifuge tubes and subsequently sonicated for 3 minutes at 70 % output at room temperature . a positive or partially positive coating result was apparent from visual inspection of the manner in which light was scattered by the sample . to determine the efficiency of coating , resultant samples were further observed for settling and also were fractionated into colloidal coated magnetite or microagglomerants thereof in the following fashion : 0 . 5 ml aliquots of the sonicated mixture in 12 × 75 mm test tubes were placed in a ciba - corning magnetic separator ( walpole , mass .) and visually observed . the criterion used to determine if the coated magnetite crystals were colloidal was to observe if the resultant material remained in solution in the magnetic separator for a period of 10 minutes , i . e . in the magnetic supernatant . this criterion was established on the basis that solutions of protein coated colloidal magnetite prepared by the process of owen et al . could not be separated magnetically in the magnetic separator when placed therein for such periods . to determine what portion of the sonicated mixtures formed stable , coated , but agglomerated materials which pulled to the sides of the test tubes in the magnetic separator , materials so formed were washed with 20 mm phosphate three times and resuspended in the same buffer . their resuspension characteristics compared with uncoated magnetite were distinctly different , as they would remain suspended for hours compared with minutes for the latter . a second criterion involved examining the magnetic supernatant as follows : putative colloidal material was separated from the mother liquor by hgms , washed , and resuspended in buffer , and visually observed for colloidal appearance and stability . hgms was accomplished by placing approximately 20 mg of fine grade stainless steel wool ( mcmaster - carr , new brunswick , n . j .) ( washed in detergent , incubated in 1 % bsa phosphate buffered saline ( pbs ), rinsed with deionized water , dried and cut in approximately 3 mm lengths ) into 12 × 75 mm glass test tubes . 100 microliters of the supernatants to be tested were added to the test tubes containing the stainless steel wool and placed in the magnetic separating rack for 2 minutes by which time magnetic material had collected on the stainless steel wires . clear non - magnetic supernatants were removed with pasteur capillary pipettes and the magnetic material washed 3 times with 300 microliters of 20 mm phosphate ( ph 7 . 5 ). after the 3rd wash , collected magnetic material was resuspended in the phosphate buffer , after removing the tube from the magnetic rack , and examined visually for the appearance of a stable colloid . in some instances recovered material was also sized by laser light scattering ( coulter sub micron particle analyzer n4sd , hialeah , fla .). some supernatants were separated from unadsorbed material by gel filtration chromatography on sephacryl - 300 ( pharmacia ) or ultra gel aca - 22 . table i lists different compounds used in coating experiments , their concentration ( s ), and other solvent conditions . the presence and semi - quantitation of colloidal and agglomerated materials were determined visually by the procedures described above , by comparison to standards prepared from known mixes of colloidal and agglomerated stocks . as can be seen from the table , every compound tested produced some amount of stable colloidal material . where noted , stability , following hgms and resuspension in phosphate buffer , is also indicated . in all cases the success of the coating experiment could be determined immediately following the sonication procedure from light scattered or by the apparent &# 34 ; shininess &# 34 ; of the solution . materials for which coating conditions were inappropriate ( i . e ., stable coated subdivided particles were not produced ) were by contrast dull and heterogeneous in appearance . table i______________________________________supernatantcoating treatment ofmaterial and colloidalconcentration percent hgms subsequent ( wt %) colloidal recovery resuspension______________________________________dextran , 25 % 100 % good gooddextran , 12 . 5 % 50 % good gooddextran , 6 . 25 % 25 % nd nddextran , 3 . 12 % 5 % nd ndsds , . 5 % 5 % nd ndsds , . 25 % 20 % nd ndsds , . 125 % 80 % good poorsds , . 063 % 50 % nd ndtween - 20 , 1 % 20 % good fairtween - 20 , . 5 % 30 % nd ndtween - 20 , . 25 % 40 % nd ndtween - 20 , . 125 % 50 % nd ndtween - 20 , . 063 % 75 % nd ndtween - 20 , . 031 % 90 % nd ndtween - 20 , 3 . 5 % 50 % nd ndbsa , 1 %; 50 mm p 95 % good goodbsa , 7 . 5 %; 50 mm p 80 % nd ndbsa , 7 . 5 %; 40 mm p 75 % nd ndbsa , 7 . 5 %; 30 mm p 50 % nd ndbsa , 7 . 5 %; 20 mm p 90 % nd ndbsa , . 25 %; 95 % good good . 016 % tween - 20igg ( human ), 5 %; 60 % good goodpbs , 50 % lipid stripped 100 % good goodhuman serag α mfc / 50 mm p 1 : 2 5 % nd ndg α mfc / 50 mm p 1 : 4 50 % nd ndg α mfc / 50 mm p 1 : 8 5 % nd ndg α mfc / 50 mm p 1 : 16 30 % nd ndg α mfc / 50 mm p 1 : 32 20 % nd ndg α mfc / 50 mm p 1 : 64 5 % nd ndsteroid free , lipid 5 % nd ndstripped sera , neatsteroid free , lipid 50 % nd ndstripped sera / 50 mm p ( 1 : 128 ) peg , 20 % 80 % nd ndgla , 10 % 100 % nd ndpoly g , 10 % 15 % nd ndpolyvinyl 75 % nd ndpyrrolidone , 15 % polyvinyl 50 % nd ndalcohol , 10 % ______________________________________ nd = not done p = phosphate buffer g α mfc = goat antimouse fc [ jackson labs , west grove , pa sds = sodium dodecyl sulfate peg = polyethylene glycol [ matheson , coleman and bell , east rutherford , n dextran = dextran t40 [ pharmacia , piscataway , nj poly g = polyglutamic acid [ nen , pilot chemical division , boston , ma gla = [( glutamic acid 45 mole %) ( lysine 35 mole %) ( alanine 20 mole %)]. sub . n [ nen , pilot chemical division , boston , ma igg = immunoglobulin g lipid stripped and steroid free lipid stripped sera from scantibodies inc ., santee , ca tween20 = polyoxyethylene surfactant ( ici americas ) i - 125 labeled bsa and igg were prepared by the iodogen method of fracker and speck ( biochem . biophys . res . comm . 80 849 , ( 1978 )). specific activities were 520 , 000 cpm / ug and 810 , 000 cpm / ug , respectively . bsa was coated onto magnetite under two sets of conditions as follows ( 1 ) at 7 . 5 % bsa in 10 mm phosphate buffer followed by 3 minute sonication and ( 2 ) at 7 . 5 % bsa in 25 mm phosphate buffer followed by 3 minute sonication . in both cases 800 , 000 cpm of labeled bsa was added to the mixture ( 0 . 5 ml ) prior to sonication . from visual inspection it was apparent that the higher concentration of phosphate resulted in significantly more colloidal material . the magnetically collectable precipitate for each experiment was collected , washed twice in 20 mm phosphate buffer and counted . the magnetic agglomerated material of conditions 1 and 2 contained 2421 and 2828 cpm , respectively . to determine how well bsa had adsorbed to these agglomerated preparations the recovered material was suspended in 0 . 1m glycine at ph 3 . 0 for 40 minutes at room temperature , magnetically separated and washed once with 20 mm phosphate buffer . for experimental conditions 1 and 2 , counts retained were respectively 91 and 84 %. when similar preparations were resuspended in buffer and allowed to incubate over night at 37 ° c ., no material desorbed from the magnetite . to determine the stability of the colloidal magnetite bsa , the amount of bsa adsorbed onto the colloid was quantified as follows : 3 × 100 ul aliquots of the supernatant were individually collected by hgms , washed 2 times with 300 ul of phosphate buffer and resuspended into 100 ul of the same buffer . by observation over time ( 16 hours ) it was apparent that the colloidal material was stable . radioactive bsa recovered by hgms for the total supernatant was 5400 cpm . from radioactivities and volumes of the supernatant and agglomerated materials obtained for condition 2 it was determined that 7 , 828 cpm &# 39 ; s had been incorporated onto magnetite . this amounts to 0 . 01 % of the total bsa added to the system which corresponds to about 0 . 75 mg / ml of magnetite so prepared . this value is very close to the optimal amount of bsa which can be coated onto magnetite by the procedure of owen et al . the igg coating experiments were done at 5 % protein concentration in pbs / 2 ( a 1 : 2 dilution of pbs in h 2 o . before sonication , mixtures were spiked with 1 . 8 × 10 6 cpm of radio labeled igg . for these experiments it was found that 1 . 2 % of the total added protein was adsorbed which corresponds to 1 . 2 mgs igg / ml of magnetite . again this is close to the maximum coating obtainable by the owen et . al . teaching . when hgms was performed on the magnetic supernatant of this experiment 23 , 000 cpm were retained on the colloidal material . the agglomerated magnetic pellet retained 17 , 300 cpm &# 39 ; s after multiple washings . when the magnetic pellet was resuspended in glycine at ph 3 . 0 as above only 50 % of the counts were retained on the subsequent magnetic pellet . however , in these experiments it was found that glycine treatment of the microagglomerated material had converted a substantial portion ( approximately half ) to colloidal material ; hence , the igg coated material is indeed stable . colloidal materials recovered by hgms were stable by visual observation . goat anti - mouse fc ( obtained from jackson laboratories , west grove , pa .) was coated onto magnetite as described in example 1 . for coating , the untreated anti - sera was diluted 1 : 4 with 50 mm phosphate . after the sonication procedure , most of the resultant material appeared colloidal . the entire supernatant was separated on sephacryl - 300 . colloidal material appearing in the void volume was recovered and the following test performed : 100 ul of the recovered colloid was mixed with either 100 , 000 counts of 125 - i labeled mouse igg or 100 , 000 counts of 125 - i bsa . these mixtures were incubated in 12 × 75 mm tubes in the presence of iron powder as described above . after 90 minutes at room temperature , hgms was performed , as described above , supernatants were discarded , and collected material washed twice with 0 . 8 mls of pbs containing 2 % bsa . for these colloidal samples ( in triplicate ) it was found on average , 5 , 200 counts of mouse igg was bound to the fc - coated colloid versus 632 cpm of nonspecifically bound bsa . mixed transition metal oxides were prepared at room temperature and at 65 ° c . by addition of base to appropriate metal chlorides as described in example 1 . table ii lists preparative conditions , molar ratios , initial color of the oxides and color after 1 week , or color after bubbling o 2 through freshly prepared oxides for eight hours . table ii__________________________________________________________________________concentration ( mmole / liter ) original finalpart . fecl . sub . 2 vcl . sub . 3 cocl . sub . 2 pdcl . sub . 3 dycl . sub . 3 ercl . sub . 3 color color * __________________________________________________________________________1 -- 5 -- -- 5 -- dk green white2 2 . 7 3 . 6 -- -- 3 . 6 -- dk green yellow3 5 3 . 3 -- -- 1 . 7 -- dk green orange4 6 -- -- -- 4 -- dk green rust5 -- -- 5 5 -- -- green brown6 -- -- -- 5 5 -- brown brown7 -- 5 5 -- -- -- blue / green green8 -- 5 -- -- -- 5 lt brown white9 -- -- -- 5 -- 5 brown brown10 -- -- -- -- 5 5 white white11 5 -- -- -- -- 5 green orange__________________________________________________________________________ * these oxides change color gradually in about one week , or in about eight hours if the new precipitate is bubbled through with oxygen . table iii lists the results of coating the oxide preparations of table ii ( preps 1 , 2 , 3 and 4 ) with dextran and with bsa , according to the method of this invention . clearly these materials can be coated similarly to magnetite . it is noted that the non - ferrite and ferrite oxides listed in table ii could be used in place of magnetite with the teaching of whitehead et al . to produce similarly colored silane coated materials . when the appropriate chlorides ( or sulfates ) of these metals are used with the teachings of molday or with owen et al ., it is possible to obtain colloidal materials which are nearly transparent in the visible region . table iii______________________________________appearance after sonicationpar - % ticlecompound supernatant pellet colloidal______________________________________1 25 % dextran shiny white white 90 % 2 25 % dextran cloudy yellow white 20 % 3 25 % dextran clear yellow white 95 % 4 25 % dextran clear orange orange 50 % 1 1 % bsa / 50 mm p shiny white white 90 % 2 1 % bsa / 50 mm p cloudy yellow white 90 % 3 1 % bsa / 50 mm p clear yellow white 95 % 4 1 % bsa / 50 mm p clear orange orange 75 % ______________________________________ bsa magnetite was prepared with the magnetite of example 1 and bsa at 1 % in 50 mm phosphate at ph 7 . 0 as described above except that an aliquot was sonicated for a second three minute period and another for a third three minute period . after removal of any agglomerated material with the magnetic rack , as described above , the resultant colloidal samples were sized by laser light scattering ( coulter submicron particle analyzer ). within experimental error each preparation had a mean particle diameter of 80 nm . next , magnetite for coating was prepared by two methods which have been found to decrease particle size in the owen et al . procedure , namely very rapid addition of base and very rapid addition of base at elevated temperature ( 65 ° c .). when bsa magnetite was prepared with a single 3 minute sonication step but with magnetite prepared according to owen et al ., the resultant colloids were both of 50 nm mean diameter . 3 . 75 % nh 4 oh was added at 0 . 6 mls / minute to a 200 ml stirred mixture of degassed hydrated ferric and ferrous chloride salts at 7 and 3 mg / ml respectively . as the ph of the initially acidic mixture approached 7 . 0 ( indicted by a color transition from dark orange to black ), 300 ul aliquots were removed , placed in 10 × 75 glass test tubes , and tested for the presence of magnetite , by placing a neodynium / iron / boron bar magnet to the side of the tube and visually observing magnetic clearing . the transition from non - magnetic or partially magnetic material to totally magnetic material occurred at ph 7 . 4 after 12 . 2 mls of base had been added . at this point , 60 ml of the mixture was removed and designated preparation a . additional base was added to the mixture to ph 8 . 9 ; a 60 ml aliquot was removed and labelled preparation b . base was further added to the reaction mixture to ph 9 . 8 and an aliquot of this material designated preparation c . magnetite preparations a , b , and c were washed 4 times with distilled water and resuspended in water such that the iron salts were at the starting concentrations of the original solution . 0 . 5 ml aliquots of the latter suspensions of a , b and c were sonicated for 3 minutes at 70 % output ( fisher sonic dismembrator ). immediately following sonication , each had the shiny appearance of colloidal magnetite ; however , all reverted to dull suspensions ( indicative of particle aggregates ) within 2 minutes . for preparation c , the transition occurred 20 - 30 seconds following sonication ; for preparation b , 20 - 50 seconds ; while preparation a required 2 minutes for this transition . when the preparations were placed in the corning magnetic rack immediately following sonication , they cleared completely from suspension in the same order as above and over the same time frame . after reaggregation , there was no discernible difference among the samples . to determine if prepared magnetites susceptibility to dissociation by sonication is altered by h + or oh 31 treatment , 0 . 5 ml aliquots of the preparation c suspension were placed in test tubes and separated from their water supernatants by magnetic separation and supernatant aspiration . magnetic pellets were then resuspended into 0 . 5 ml of either dilute hc1 ( 0 . 1 , 0 . 01 , 0 . 001 , or 0 . 0001m ) or dilute naoh ( 0 . 1 , 0 . 01 , 0 , 001 , or 0 . 0001m ), sonicated for 3 minutes as above and visually inspected . for these samples , it was apparent that acid or base treatment increased the disruption of magnetite in proportion to concentration of acid or base . the aliquots resuspended and sonicated in 0 . 1m hc1 or naoh remained shiny , i . e . colloidal , for nearly 12 hours . the aliquots resuspended in 0 . 001m acid or base also exhibited colloidal behavior for significant periods of time . on the other hand , aliquots resuspended into 0 . 0001m acid or base quickly aggregated after sonication much like the parent material did in water . preparation c of example 6 was coated by disruption via sonication with anionic and cationic polypeptides . an anionic terpolypeptide composed of 60 mole % glutamic acid , 30 mole % alanine , 10 mole % tyrosine ( gat , lot # m18g ) and a cationic copolymer of 60 mole % lysine , 40 mole % alanine ( la , lot # m - 5b ), both obtained from pilot chemicals , watertown , mass ., were used . these polypeptides , both of about 100 , 000 daltons , were solubilized by tituration with appropriate acid or base , neutralized , dialyzed versus phosphate buffered saline ( ph 7 . 0 ) and subsequently against distilled water . coatings were attempted with 10 mg per / ml solutions of polypeptide and serial dilution of same to 1 / 16 . coating experiments were done by sonication as described above . for these experiments , the 1 / 16 and 1 / 8 dilutions of both gat and la produced stable colloidal solutions . the 1 / 4 , 1 / 2 and undiluted polypeptide solutions resulted in distributions of colloid and agglomerated material . the amount of colloid in both instances generally decreased with increasing polypeptide concentration . however , at 25 mg / ml gat all of the material was colloidal . preparations a and c of example 6 were coated with gat . 0 . 5 ml aliquots of magnetite were resuspended in 1 . 0 ml of hcl ( 0 . 01 and 0 . 1m ) or naoh ( 0 . 01 and 0 . 1m ) for 60 seconds followed by two 0 . 5 ml water washes . these pre - treated magnetites were resuspended in 0 . 5 ml aliquots of 1 mg / ml gat containing 2 . 5 × 10 6 cpm 125i gat ( radio - labeled by the iodogen method described above ) and sonicated as described . sonicated samples were placed in the corning magnetic rack and left to separate overnight . magnetic pellets were washed once in 0 . 5 . m nacl and counted . counts bound and percentages of gat coated onto the various magnetites are given in table iv . table iv______________________________________effect of h . sup .+ / oh . sup .- treatment of magnetitepreparations a and c on gat coating preparation a preparation ctreatment cpm bound % bound cpm bound % bound______________________________________0 . 10m hcl 450 , 000 35 340 , 950 260 . 01m hcl 287 , 602 22 226 , 250 17h . sub . 2 o control 209 , 165 16 236 , 042 180 . 01m naoh 130 , 728 10 n . d . n . d . 0 . 010m naoh 90 , 600 8 54 , 010 5______________________________________ n . d . = not done from the data of table iv , it is clear that magnetite crystal surface charge can be made more positive by a pre - treatment with acid and conversely , treatment with base generally reduces sites available to the negatively charged polymer . when 125 i preparation of the cationic terpolymer lat ( 60 mole % lys , 30 mole % ala , 10 mole % tyr ), ( received from dr . h . j . callahan , jefferson medical college , philadelphia , pa .) was used with similarly pre - treated magnetite , it was observed that greater binding of this positively charged material occurred on magnetite pre - treated with base . to determine if crystal surface charged alteration via acid or base treatment is a general property of transition element oxides , such as those prepared from different molar ratios of chlorides of fe ( ii ), dy ( iii ), v ( iii ), such materials were treated with acid or base as above and mixed with either radio - labeled gat and / or salmon sperm dna ( sigma chemical co ., st . louis , mo .). as in the case with magnetite , acid treatment promoted binding of these negatively charged polyelectrolytes . in fact , uncoated materials pretreated with 0 . 1m hcl , followed by water wash and sonication give materials which behaved like preparations of magnetite described above , i . e ., transiently stable colloids . when these materials were in this semi - stable colloid state , addition of salmon sperm dna in appropriate quantities lead to complete agglutination indicating the crystals had assumed substantial positive surface charge .