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Studies with medicated water-in-oil emulsions
STUDIES
WITH
MEDICATED
I. The Use of a High-Speed Sterile Conditions ,‘amrl,cl
d.
l+ignl,
MJ).,
WATER-IN-OIL
EMULSIONS
Shaker for the Production of Emulsions Under
6’.,4.C.P.,
h’c?W
York
City,
N.
1’.
injection therapy, for t,hc treatment of pollinosis, which has been intcrmittcntly explored for many years has recently stimulated great interest and actirit,y among allergists following the reports of E. A. Brown.1-3 In order to insure safety in the USC of potentially hazardous agents, emulsification of a high order must take place so that. none or little of the aqueous phase remains uncmulsificd. Since it is also essential that the emulsions to be injected be produced under sterile conditions, a number of methods, utilizing syringes and needles in a closed system, have been advocated. These have ranged from the simple pumping by hand with the needle immersed into the liquids to be emulsified, to the use of automatic mwhines for transferring the liquids frorn one syringe into another through a double-hubbed needle. The pumping by hand technique, originally advocated I))- Freund and Tllomson,4 was the method employed by Loveless.” An improrcd method was later dcscribcd by Berlin and MeKinney” v-ho used a double syringe technique in which pressure by hand forced the fluids from one syringe to the other via a double-hubbed needle. Brown originally emplo:-ed this method but subsequently devised an electric emulsor based on the silm(J principle. His machine enabled him to use greater pressure and finer bawd needles which resulted in finer emulsions. I, in evaluating various ways of makin, v emulsions, have found that the agitation (and shear) produced by a high speed shaker, described below, made twcllent cmnlsions. ‘l‘hr apparatus is simple to operate and emulsifies in fractiorls of a minute to w\-vral minutes, depending on the materials to be emulsified, their ~olumc~, the cmnlsificrs employed, and the degree of emulsification desired. For the pwposc on hand-elnulsificatiorl of pollen antigens-agitation of volnrncs from 1 to 10 1111. for 1 or 2 minutes will produce stable emulsions in which thv particles aw rclativ(lly uniform in size and range from approximately 0.5 to 2 p.
R
EPOSITORY
The dcviw described bc~low is a larger and sturdier Bug shnkcr commonly employ-cd by dentists in preparing Avenue
Front tht: Depar’tnrent of Mellicinc (Allergy), Huspitals, New York City, N. Y. Jiweive,l for publication Feb. 1, 19fi2. *Cwscrnt Dental Manufacturing Co., 18X9 So. 295
New
Pulaski
York
Rd.,
version of the Wig-Lamalgams.* Although
Medical
Chicago,
College,
Ill.
Flower-Fifth
296 the small Wig-L-Bug shaker was originally cmployved in this investigation and prwduced satisfactory emulsions, therae wrc freyucwt brtvrkdo~\ns, and :I sturdier device was sought and found in the ter’gcr \Yig-IA-Bug (scrics 6000). The same principle is criiplo~~~l in both machinw A high-speed motor turxs ii shaft on rvhich thrrc is a mc;unt and cradlr holding the rrlatc~rials to be agitatctl. The owillatory action is \-cry rapid, appr’oximately 3,200 r.p.m. when fully toadcd. The shaft is corrncctcd with au ecccntriv bushing, producing motion along three mutually pcrpc>ndicular* ases which :rssurw rxpitl ant1 thorough mixing.
The emulsions may be made in small yuantitics for cnch patient, in whatever dosage may bc dcsiwd, or may bc pwparcd in bulk up to 10 C.C. and then transI’erred to tubc~cutiri syringes (glass or disposable plastic) in appropriate amounts for inject,ion. For the preparation of individual doses, 1 have used holders fabricated from a light polyethylene plastic into which a syringe, 1 or 2 ml. capacity, is placed and held firmly with rubber bands while being shaken. Bulk emulsions may be made in vials or in syinges of 5 and 10 ml. capacities. Thcl former has the ad\-antagc of simplicity of assembly but, suffc~s from disadv:rntagcs in use in that it is difficult to withdr*a\v the viscid emulsion, and tlrew is wustqy. Making the cmutsion in syringes, or1 the other hand, has dccidcd advantagw. Stclrility is more assurctl since tlrclrx: is less opportunitp for contamination by i’rcqucnt withdrawals, as in tilt casc~of vials. Fnrtlicmmore, the transfer of wyuirwl dosage from the Iargv syririgc into smaller syringes is readily effected yia a donbtr-hubbed netdlv. The 10 ml. l,uer-Idok irrtc~~clrarrg~~a~)le syringes a w most, suitable for the making of ttrri emulsions, althoagh srnallcr 011(‘s can also hc used. The barrel The barrel is then and plunger ~II’C scparatcd and stcrilizcd by antwtuving. closed off at both (lrrds-irt tlrc tip b>- a mcltal scwwcap locking into tlrcl I,uerLok, and at tlw open end b,v an “army typcl” ncoprwrc cap jvhich plugs into the syringe, waling it tightly. ( )nly rwoprwc~ shoiitd be used, since oils tlisintcgrate natural ruhbc~r. Tho syringe barrel now wrves as a sttbrilc wwptarlc, first for the aqueous phaw transfcrrcd in mcasurwl amounts (5 ml. COYthe 10 ml. syringe) undcv sterile techniqw via s,vinge and needle thrwugh the ruhhcr stopper. To this ma>- bc added ferric chloride to enhanw c~rrrulsificat,ion, and to test for degree of cmutsification and prcsencc of free uncmulsified ayueous phase.’ I+‘ivc millititcr3 of thv oil, pwviously ~)lY’I)illY’tl with 011~’or rnore emulsifiers, is then irijcctrd into thr SilIll? barrel cwntairiing tllc aqurous ~IllilW. There awe then present 8 distinct phases: \Vilt(‘r ilt thv bottom, the oily layer above that, with air filling the rest of the barwl. That air spavo is essential since, in the shaking prwcss to fotlow, space for agitating the liquids must bc present. Although air* is thus trapped in the cmnlsiotr, it is nttimatcIy released and removed from the emulsion, as dcwrihcd later. Alcohol on ir wtt,on swab is used to sterilize the rubber cap before each penetration, care being taken to allow the alcohol to evaporate completely before
Fig. syringe. ptl;lses
I.-Before Em?c7sificntion. A. \Vig-L-Bug machine with The syringe barrel Ins bcwn converted into a container (see text). h’. Il;sertion of barrel into carriage.
cradle for holding holding the oil
a 10 ml. an<1 water
For* storqy ilnd trxnsfrr \\hilc rrr;tintairring stclrility, :I double-hubhed nwdlc is tlwn lockrtl into tlrv s\~ringc, the open vntl pwmitting the insertion of the tip of ir tnbc~i~culin (or* other* small ) syririgr, into \vhich is trxnsferred thv emulsions, the open end cf the doublri~equirwl dew. Wl 1vn not trxnsfcrring hubhed needle is closed by. a l~u(lr-l~ok eirl). Thv syringe with its attached doublc-hubbcd nccdl~~ and protcctiv(x I,uer-Idok C;I~ may then be stored in the YCIn t,his way, emulsions may he frigcrator until another trxnsfvr is desired. rei’rigeratd for months and used safely without any need to w-emulsify, prochlwidc spot test is ncgativc~. vided tlrcl fcuic It, should 1~ cmphasizcd that, machinw do not, mnkc emulsions-they merely
298
Fig.
help in emulsification. No rnarhinc van emulsify materials not properly prcpared for cmulsitication. The vase of cmulsitication and the stability of an emulsion is primarily determined by the ph?-sicoclrc~mic;l processes acting at, the surfaces of the two phases. \\‘hcn the proper balance of the phases and emulsiticw is achicwd, crnnlsific~ation rrra~-c’vvrr occrw spontaneously. With the present materials emplo\-cd by the allergist and with his lirnitc>d knowledge of the chernistry in\-oil-cd in c,trrrrlsificatiorr, a machine fora cmulsitication is still a ncccssary adjunct
Fig.
4.-Stol’nge rind Tmnsfer of Emulsior~ A, The neoprene stopper has been removed plunger reinserted. The screw c&p at the tip has been replaced by a double-hubbal for facilitating thtl transfer of the emulsion. For storage, the open em1 of the doubteneedle is locked with a screw cap. B. For transfer, the screw cap is rernove,l muI a plastic disuosabte tuberculin syringe is inserted. Traction cn the plastic plunger, assisted by slight ~I’CSSUI’O on the glass plunger, rearlily fills the syringe to the clesire(l quantity.
an<1
the
needle hubbed
Although emulsions can 1~ made by hand am1 ha\.e been prepawd in this fashion by I~owIcss,” tliq- cannot consistrntly duplicate nraclrino-made emulsions. The latter arc more uniform in particle-size distribution and more st,able on standing. Figs. 1-4 illustrate the proccdurw of emulsification, storage, and transfer of allcqyns.
Gross inspection permits only the drtcction of obvious failnw tion, such as the complctc separation of the two phases or partial
of emulsificaemulsification
300
1’K IGAI,
J. Allergy .l,,ly-
~A,,~nat. 1962
in which there arc hwaks in homogmcity. I~:\Y~IIw11~w an emulsion looks winpletely lio~noge~icous 1)~ gims inspcrtion, it is not yt safe to cmplo~ sinco it dots not indicate whcthw it is ii water-in-oil or oil-in-water (invc~rsion) mnulsion ; nor dots it indicate the de~/wc of cmulxification. ISarlicxr writers lia~c~ c~xprtssed the dcgrcc of eninlsification 1)~ thv ” fwI ” of it-it should feel smooth and silky-w its adequacy l)>- tlroJ)ping on water ilrld Iloting whcthw the cnlr~lsior~ dlaoplct floats.” Ncithcr c;f thwc m&hods is wliahlc (wough for WV I)>- the illlcrgist. It is prefcrahlc to c~sainirw tlicl cwulsion niiv twwopic7ll ly I)y rc~mo\-ing a drop of the emulsion with the cdgc of iI toothpic!: and tr;l~lsf(~rri~lg it to it drop of rninwal oil previously plawd 011 glass slide. This is thc;mughly noised, (dov(‘rcd with iI COYPYSliJ), J>lYSSC’d fir~nly, ant3 t,licn cw~lnined for tlroJ)Ic~t size and lioniogcnc~ity. I)ropping the c~rnulsion on wa.tcr tlocv wl’w to tliffc~lYIltiiltc~ tKtown wat,er-in-oil and cil-in-water rrnulsions-the formc~, if wtll Inad(b. will float and remain as an intact globule, \~ller(~as ihc latter will spwad :rntl disintcyratc since, in thtk invcrtcd (xmulsion, tllo ontrr ~)llitSO (\ViltP1') is frwl~dissiJ)atcvl. Thcw arc other more ~c+inctl tncthods for dctwting the degwc and type of cniulsification-t~l(~~~t~~i~~~l, cticinic~al, and biologic. Thcsc prowdurcs arc nmlwing used 11.~mc in comJ)aring the various nwthcxls o I’ Itraking c~mulsions and will 1~1reported on. il
11 high-spwd shaking dcvive is descritwd for tlio ~mrposcs of making ornulsions rapidly under stcrilr conditions. This lcwds itself particularly to the prepilratioll of emulsified allcrgvns for wpositoi~y (single dow) treatnicnt 01’ hay fcrcr and related discascs. The author wishes t,o acknowledge the help provided 1)~ Mr. John S’c~enga, rncc-llanio, New York Mrdicnl (lollegc, l)vpartnwlrt of Physiology, 11y Mr. Morton Mc~orac~ll of Allied Plsstirs, New Pork City, ant1 1)~ Mr. Ect\vard Sheridan of (!rescent Dental Rlanufacturing PO., Chicago, III. Mr. Irving Rprvlman of thcl Proppcr Mxnufac*turing (:o., 10-3-l 44t,h l)rivp, Long Island City 1, N. Y., providrd the sczrew raps and tlouldc-hul)l~d nrcdl~.
Since this report has developed a simple 5, or 10 C.C. capacities, emulsification procedure.
I. Brown, 2. 3. 4. 5. 6. 7.
was sulnnittetl for pul)lic:ttiorl, the manufacturer of the apparatus metal clamp capaljle of holding 111 CC’. vials or syringes of 1, 2, theret,! elirniuating the plastic holder and simplifying further the
E. A. : The Treatmrnt of Pollinosia IIV Illcans of a Single dnnuxl Injcvtion of Emulsified Extract. V, Ann. Allrrgy 17: 34, 1959. Brown, E. A. : The Treatment of Allergy to Ragw~(l I’olleu by Means of a Single Annual Injection of Emulsified Pollen Extract. VI, Ann. Allergy 17: 358, 1959. Brown, E. A.: Treatment of Grass Pollinosis With a Single Annual injection of Emulsified Extract. IV, Ann. Allergy 16: 510, 195% Freund, J., and Thomson, K. J.: A SimpltL Rapid Trc*lmiyuc~ of Prc~pariug U’ater-in-Oil Emulsions of Penicillin, Drugs and BiologicY, Hcicnvcl 101: 468, 1945. Loveless, M. H.: Repository Injection in Pollen Allergy, .J. Immunol. 79: 68, 1957. A Simple l)cvivc for Making Emulsificvl Vxcrinvs, Berlin, B. S., and McKinney, R. W.: J. T,ah. & Clin. Med. 52: 657, 1958. Priyal, S. J.: Improved Emulsions of Allergens With Built-in Safety Features, New York State J. Med. 61: 3615, 1961.
WITH
MEDICATED
I. The Use of a High-Speed Sterile Conditions ,‘amrl,cl
d.
l+ignl,
MJ).,
WATER-IN-OIL
EMULSIONS
Shaker for the Production of Emulsions Under
6’.,4.C.P.,
h’c?W
York
City,
N.
1’.
injection therapy, for t,hc treatment of pollinosis, which has been intcrmittcntly explored for many years has recently stimulated great interest and actirit,y among allergists following the reports of E. A. Brown.1-3 In order to insure safety in the USC of potentially hazardous agents, emulsification of a high order must take place so that. none or little of the aqueous phase remains uncmulsificd. Since it is also essential that the emulsions to be injected be produced under sterile conditions, a number of methods, utilizing syringes and needles in a closed system, have been advocated. These have ranged from the simple pumping by hand with the needle immersed into the liquids to be emulsified, to the use of automatic mwhines for transferring the liquids frorn one syringe into another through a double-hubbed needle. The pumping by hand technique, originally advocated I))- Freund and Tllomson,4 was the method employed by Loveless.” An improrcd method was later dcscribcd by Berlin and MeKinney” v-ho used a double syringe technique in which pressure by hand forced the fluids from one syringe to the other via a double-hubbed needle. Brown originally emplo:-ed this method but subsequently devised an electric emulsor based on the silm(J principle. His machine enabled him to use greater pressure and finer bawd needles which resulted in finer emulsions. I, in evaluating various ways of makin, v emulsions, have found that the agitation (and shear) produced by a high speed shaker, described below, made twcllent cmnlsions. ‘l‘hr apparatus is simple to operate and emulsifies in fractiorls of a minute to w\-vral minutes, depending on the materials to be emulsified, their ~olumc~, the cmnlsificrs employed, and the degree of emulsification desired. For the pwposc on hand-elnulsificatiorl of pollen antigens-agitation of volnrncs from 1 to 10 1111. for 1 or 2 minutes will produce stable emulsions in which thv particles aw rclativ(lly uniform in size and range from approximately 0.5 to 2 p.
R
EPOSITORY
The dcviw described bc~low is a larger and sturdier Bug shnkcr commonly employ-cd by dentists in preparing Avenue
Front tht: Depar’tnrent of Mellicinc (Allergy), Huspitals, New York City, N. Y. Jiweive,l for publication Feb. 1, 19fi2. *Cwscrnt Dental Manufacturing Co., 18X9 So. 295
New
Pulaski
York
Rd.,
version of the Wig-Lamalgams.* Although
Medical
Chicago,
College,
Ill.
Flower-Fifth
296 the small Wig-L-Bug shaker was originally cmployved in this investigation and prwduced satisfactory emulsions, therae wrc freyucwt brtvrkdo~\ns, and :I sturdier device was sought and found in the ter’gcr \Yig-IA-Bug (scrics 6000). The same principle is criiplo~~~l in both machinw A high-speed motor turxs ii shaft on rvhich thrrc is a mc;unt and cradlr holding the rrlatc~rials to be agitatctl. The owillatory action is \-cry rapid, appr’oximately 3,200 r.p.m. when fully toadcd. The shaft is corrncctcd with au ecccntriv bushing, producing motion along three mutually pcrpc>ndicular* ases which :rssurw rxpitl ant1 thorough mixing.
The emulsions may be made in small yuantitics for cnch patient, in whatever dosage may bc dcsiwd, or may bc pwparcd in bulk up to 10 C.C. and then transI’erred to tubc~cutiri syringes (glass or disposable plastic) in appropriate amounts for inject,ion. For the preparation of individual doses, 1 have used holders fabricated from a light polyethylene plastic into which a syringe, 1 or 2 ml. capacity, is placed and held firmly with rubber bands while being shaken. Bulk emulsions may be made in vials or in syinges of 5 and 10 ml. capacities. Thcl former has the ad\-antagc of simplicity of assembly but, suffc~s from disadv:rntagcs in use in that it is difficult to withdr*a\v the viscid emulsion, and tlrew is wustqy. Making the cmutsion in syringes, or1 the other hand, has dccidcd advantagw. Stclrility is more assurctl since tlrclrx: is less opportunitp for contamination by i’rcqucnt withdrawals, as in tilt casc~of vials. Fnrtlicmmore, the transfer of wyuirwl dosage from the Iargv syririgc into smaller syringes is readily effected yia a donbtr-hubbed netdlv. The 10 ml. l,uer-Idok irrtc~~clrarrg~~a~)le syringes a w most, suitable for the making of ttrri emulsions, althoagh srnallcr 011(‘s can also hc used. The barrel The barrel is then and plunger ~II’C scparatcd and stcrilizcd by antwtuving. closed off at both (lrrds-irt tlrc tip b>- a mcltal scwwcap locking into tlrcl I,uerLok, and at tlw open end b,v an “army typcl” ncoprwrc cap jvhich plugs into the syringe, waling it tightly. ( )nly rwoprwc~ shoiitd be used, since oils tlisintcgrate natural ruhbc~r. Tho syringe barrel now wrves as a sttbrilc wwptarlc, first for the aqueous phaw transfcrrcd in mcasurwl amounts (5 ml. COYthe 10 ml. syringe) undcv sterile techniqw via s,vinge and needle thrwugh the ruhhcr stopper. To this ma>- bc added ferric chloride to enhanw c~rrrulsificat,ion, and to test for degree of cmutsification and prcsencc of free uncmulsified ayueous phase.’ I+‘ivc millititcr3 of thv oil, pwviously ~)lY’I)illY’tl with 011~’or rnore emulsifiers, is then irijcctrd into thr SilIll? barrel cwntairiing tllc aqurous ~IllilW. There awe then present 8 distinct phases: \Vilt(‘r ilt thv bottom, the oily layer above that, with air filling the rest of the barwl. That air spavo is essential since, in the shaking prwcss to fotlow, space for agitating the liquids must bc present. Although air* is thus trapped in the cmnlsiotr, it is nttimatcIy released and removed from the emulsion, as dcwrihcd later. Alcohol on ir wtt,on swab is used to sterilize the rubber cap before each penetration, care being taken to allow the alcohol to evaporate completely before
Fig. syringe. ptl;lses
I.-Before Em?c7sificntion. A. \Vig-L-Bug machine with The syringe barrel Ins bcwn converted into a container (see text). h’. Il;sertion of barrel into carriage.
cradle for holding holding the oil
a 10 ml. an<1 water
For* storqy ilnd trxnsfrr \\hilc rrr;tintairring stclrility, :I double-hubhed nwdlc is tlwn lockrtl into tlrv s\~ringc, the open vntl pwmitting the insertion of the tip of ir tnbc~i~culin (or* other* small ) syririgr, into \vhich is trxnsferred thv emulsions, the open end cf the doublri~equirwl dew. Wl 1vn not trxnsfcrring hubhed needle is closed by. a l~u(lr-l~ok eirl). Thv syringe with its attached doublc-hubbcd nccdl~~ and protcctiv(x I,uer-Idok C;I~ may then be stored in the YCIn t,his way, emulsions may he frigcrator until another trxnsfvr is desired. rei’rigeratd for months and used safely without any need to w-emulsify, prochlwidc spot test is ncgativc~. vided tlrcl fcuic It, should 1~ cmphasizcd that, machinw do not, mnkc emulsions-they merely
298
Fig.
help in emulsification. No rnarhinc van emulsify materials not properly prcpared for cmulsitication. The vase of cmulsitication and the stability of an emulsion is primarily determined by the ph?-sicoclrc~mic;l processes acting at, the surfaces of the two phases. \\‘hcn the proper balance of the phases and emulsiticw is achicwd, crnnlsific~ation rrra~-c’vvrr occrw spontaneously. With the present materials emplo\-cd by the allergist and with his lirnitc>d knowledge of the chernistry in\-oil-cd in c,trrrrlsificatiorr, a machine fora cmulsitication is still a ncccssary adjunct
Fig.
4.-Stol’nge rind Tmnsfer of Emulsior~ A, The neoprene stopper has been removed plunger reinserted. The screw c&p at the tip has been replaced by a double-hubbal for facilitating thtl transfer of the emulsion. For storage, the open em1 of the doubteneedle is locked with a screw cap. B. For transfer, the screw cap is rernove,l muI a plastic disuosabte tuberculin syringe is inserted. Traction cn the plastic plunger, assisted by slight ~I’CSSUI’O on the glass plunger, rearlily fills the syringe to the clesire(l quantity.
an<1
the
needle hubbed
Although emulsions can 1~ made by hand am1 ha\.e been prepawd in this fashion by I~owIcss,” tliq- cannot consistrntly duplicate nraclrino-made emulsions. The latter arc more uniform in particle-size distribution and more st,able on standing. Figs. 1-4 illustrate the proccdurw of emulsification, storage, and transfer of allcqyns.
Gross inspection permits only the drtcction of obvious failnw tion, such as the complctc separation of the two phases or partial
of emulsificaemulsification
300
1’K IGAI,
J. Allergy .l,,ly-
~A,,~nat. 1962
in which there arc hwaks in homogmcity. I~:\Y~IIw11~w an emulsion looks winpletely lio~noge~icous 1)~ gims inspcrtion, it is not yt safe to cmplo~ sinco it dots not indicate whcthw it is ii water-in-oil or oil-in-water (invc~rsion) mnulsion ; nor dots it indicate the de~/wc of cmulxification. ISarlicxr writers lia~c~ c~xprtssed the dcgrcc of eninlsification 1)~ thv ” fwI ” of it-it should feel smooth and silky-w its adequacy l)>- tlroJ)ping on water ilrld Iloting whcthw the cnlr~lsior~ dlaoplct floats.” Ncithcr c;f thwc m&hods is wliahlc (wough for WV I)>- the illlcrgist. It is prefcrahlc to c~sainirw tlicl cwulsion niiv twwopic7ll ly I)y rc~mo\-ing a drop of the emulsion with the cdgc of iI toothpic!: and tr;l~lsf(~rri~lg it to it drop of rninwal oil previously plawd 011 glass slide. This is thc;mughly noised, (dov(‘rcd with iI COYPYSliJ), J>lYSSC’d fir~nly, ant3 t,licn cw~lnined for tlroJ)Ic~t size and lioniogcnc~ity. I)ropping the c~rnulsion on wa.tcr tlocv wl’w to tliffc~lYIltiiltc~ tKtown wat,er-in-oil and cil-in-water rrnulsions-the formc~, if wtll Inad(b. will float and remain as an intact globule, \~ller(~as ihc latter will spwad :rntl disintcyratc since, in thtk invcrtcd (xmulsion, tllo ontrr ~)llitSO (\ViltP1') is frwl~dissiJ)atcvl. Thcw arc other more ~c+inctl tncthods for dctwting the degwc and type of cniulsification-t~l(~~~t~~i~~~l, cticinic~al, and biologic. Thcsc prowdurcs arc nmlwing used 11.~mc in comJ)aring the various nwthcxls o I’ Itraking c~mulsions and will 1~1reported on. il
11 high-spwd shaking dcvive is descritwd for tlio ~mrposcs of making ornulsions rapidly under stcrilr conditions. This lcwds itself particularly to the prepilratioll of emulsified allcrgvns for wpositoi~y (single dow) treatnicnt 01’ hay fcrcr and related discascs. The author wishes t,o acknowledge the help provided 1)~ Mr. John S’c~enga, rncc-llanio, New York Mrdicnl (lollegc, l)vpartnwlrt of Physiology, 11y Mr. Morton Mc~orac~ll of Allied Plsstirs, New Pork City, ant1 1)~ Mr. Ect\vard Sheridan of (!rescent Dental Rlanufacturing PO., Chicago, III. Mr. Irving Rprvlman of thcl Proppcr Mxnufac*turing (:o., 10-3-l 44t,h l)rivp, Long Island City 1, N. Y., providrd the sczrew raps and tlouldc-hul)l~d nrcdl~.
Since this report has developed a simple 5, or 10 C.C. capacities, emulsification procedure.
I. Brown, 2. 3. 4. 5. 6. 7.
was sulnnittetl for pul)lic:ttiorl, the manufacturer of the apparatus metal clamp capaljle of holding 111 CC’. vials or syringes of 1, 2, theret,! elirniuating the plastic holder and simplifying further the
E. A. : The Treatmrnt of Pollinosia IIV Illcans of a Single dnnuxl Injcvtion of Emulsified Extract. V, Ann. Allrrgy 17: 34, 1959. Brown, E. A. : The Treatment of Allergy to Ragw~(l I’olleu by Means of a Single Annual Injection of Emulsified Pollen Extract. VI, Ann. Allergy 17: 358, 1959. Brown, E. A.: Treatment of Grass Pollinosis With a Single Annual injection of Emulsified Extract. IV, Ann. Allergy 16: 510, 195% Freund, J., and Thomson, K. J.: A SimpltL Rapid Trc*lmiyuc~ of Prc~pariug U’ater-in-Oil Emulsions of Penicillin, Drugs and BiologicY, Hcicnvcl 101: 468, 1945. Loveless, M. H.: Repository Injection in Pollen Allergy, .J. Immunol. 79: 68, 1957. A Simple l)cvivc for Making Emulsificvl Vxcrinvs, Berlin, B. S., and McKinney, R. W.: J. T,ah. & Clin. Med. 52: 657, 1958. Priyal, S. J.: Improved Emulsions of Allergens With Built-in Safety Features, New York State J. Med. 61: 3615, 1961.