Downloaded via INDIAN [NST OF TECH KHARAGPUR on August 9, 2022 at 16:08:20 (UTC). imately sate published articles, Stability of Colorimetric Reagent for Chromium, s-Diphenylcarbazide, in Various Solvents PAUL F, URONE Division of Industrial Hygiene, Ohio Department of Health, Columbus, Obie ‘The deterioration and sensitivity of s-diphenslearbazide were studied in a variety of solutions involving seven ‘organic solvents. Nonaqueous ethyl acetate and nee- tone solutions were stable for months; methyl ethyl ketone, methyl Cellosolve, and 2-propanol solutions were usable for 1 to 2 weeks. Aqueous solutions and solvents tending to be hasic, such as methanol, ethyl alcohol, and those containing traces of water oF basi impurities, do not make good solvents for stock solu- tons of the colorimetric reagent, WIDELY used colorimetric agent for chromium is diphenslesrbazide (1,5-diphenslacarbohydrazide). How ver, there hse heen uo standard raethod of preparing « solution of thie reagent. Cazeneuve (8) used the powdered compound oF fn acetie acid solution; Moulin (8) used! an acetic ackt-ethy! Alcohol solution; Rowland (9) and Feil (6) used ethy} aleohol solutions; Graham (7) used eulfuri aed-ethy1uleohol solution; ‘and Sandell (14) used a 1 to 1 cetonewster solution, Davis ‘and Baoon (4) briefly discussed the sensitivity and life of the re gent stating that slooho! solutions lose sensitivity within 2 ddaye, aetie acid solutions deteriorate rapidly, and aoetone ol tions seemed to hold up for longer periods of time, As a rule iments were to be taken, ‘Mare recently Ege and Silverman (5) proposed a stable solution in which 0.25 gram of sdiphonylearbazide and 4.0 grame of phthalic anhydride are ditolved in 100 ml of 959% eth aloohel ‘Tho reagent was usuble for much longer periods of time and was ‘adaptable fr filter paper impregnation (12). During the course of » recent chromium study (13) 2 1 t0 1 fcetone-eater solution of the colorimetric reagent was used. Ie was noticed that analytical quality acetone irom diferent cheinieal companies give solutions of varying stability. The least stable were solutions made with acetone having a slightly hase reaetion, These would darken within a short time snd when reacted with sexivalet chromium would give low color ingensities, Hoever, solutions made from acetone having a slightly seidie restion were usuble for Tonger periods of time. ‘The chemiest ompanies concerned attributed the boesicty to traces of ammonia and the acidity to traces of propionic seid, ale though traces of formic and acetic acids tre more probable, EXPERIMENTAL To study the phenomenon further, duplieate solutions of dipherylesrbaside in various typos of asetone wore mde. ‘One set of the solutions was Kept in 6 teltigerstor at 4" C. Tt was exposed to Tight only for the short Lime necessary fo tntdean” Sen portions “of eal Semple, These portions were Used for ‘eating the abeorbnce of the {esol following which 0.5 onl of ese portion way added to'5'y of fexivalent Bhromiom. in 10.0 mL of G2N sulfur eld ad the reeuting eolgr waa meas ted. "The unused part of the portions tas discarded and aot returned tothe org container The dapat at wa uid alls ut ‘vas kept ontop of reagent shelf cleanly exposed to room light Lionel) eh tempemtares Readings weve tlken at hoy Intervals for the frst © hours, daily for the St week and weekly {Hereafter Later, tolutions of she eolorimetrio reagent im six ber organic solvents were studied inthe same manner. In al, Sorte Sb solutions were sutied. “Ai solutions were 0.297% in t-tiphenylearbazio, and ll mete- urements were made with a Becksnan Model DU spectrophotem= wierat odd and ung tn ele oid! ders {ones were measured with a Beekvsan pit meter, However, the Siphensearbaaie fw even an an tat sg Boe solutions i assumes fs base color nd curs light eel within an four or two, The sede solutions remain aighly yellow oF Coloreas unt deterioration takes pce DISCUSSION ‘The stability of the solutions of the diphenslesrbaxide was measured in term of the absorbance of the discoloration (or darkening) developed by the solution with time and the absorbance of the chromium diphenglearbazide color ob- tained when 0.5 mil of the same solution was reaeted with 3 of sexivalent chromium in 10 ml of O2N sulfuric acid. Figures Tand 2 give curves typical of the remults obtained from the tunrfrigerated samples. The reftigersted samples gave slighty Detter results, but they followed the sane genera patterns a9 their nnweignratnd cipliontes. Tf drawa, the ures of the refrigerated samples would closely parallel those of their teepec- tive tnrerigerated samples for the rt few days, being displaced slightly to the right. In 1 to 2 weeks the diferences would hee come smal, and the curves would tend to merge. Hence the Beneral pattern of stability are dependent more upon the txpe of solution than they are on refigeration Discoloration of Diphenslearbazide Solutions. When freshly made, the diphenytearbaride solutions are water white to slighty yellow in colon. As the slutions age, they become yellow-broxD to orange-red depending on the type of solvent andthe amount of deterioration. Presumshly, the diphenglearbazide i oxidized tothe esrbazone. However the reaction isnot completely under- stood. Sandell (10) states that sexivalent chromium doss not react mith pure diphenslearbszone while Bose (2) states that it sroens 230, iarog uisiey aa ETO.VOLUME 27, NO. 8, AUGUST 1955 / “wwe mbar Figure 2. Effect of time on sensitivity of s-diphenylearbazide solutions does, In any event, the reagent does lose sensitivity as i diss with excess reagent. color. ‘The 1 to 1 aqueous solution of the colorimetric reagent cade with analytical quality acetone having # slightly basic reaction ve, Figure 1) discolored rapidly, due possibly to a basic of the oxidation of the diphenslearbaside to the diphenylearbazone (J4). The ssme type of solstion, using lacetones which conformed to National Formulary ancl AMERICAN Comanicar Socrery specifications and whieh had slightly acidie reactions dd not discolor rapidly for the first few days, Hon fever, after this intial period they deteriorated at approximately the sane rate ‘Nonaqueous solutions proved to have greater stability. A olution of the colorimetric rengent in acetone conforming to ACS specifications (elightiy acidic) wae stable for months ‘A solution (not shown) prepared from acetone, purified by drying over anhydrous calcium sulfete, followed by distillation from phosphorus pentoxide (1), and Kept in s refrigerator, discolored only slighty and gave setisfctory results after a2 year period Ethyl acetate also made a slution which was stable and Gssble for as much a 18 monthe after the period of study shown here, Methanol soltions were ineatiefactory, while ethyl aleokol sale tions could be uae for approximately a week. Solutions with rethylethy! ketone, propanol, and methy] Celloslve ethylene lvcol monomethy! ether) were usable for approximately 2 weeks, Except for acetone, no attempt ras made to obtain a high purity” produet, The avetone having ACS specications and having a slightly basis reaction wa redistlled, but a center face tion guve essentially the aame resulta, The eddition of a drop fof OV wolfurie tckl increased the stability somewhat, but the discoloration followed the goncral pattern ofthe aquoots acetone ‘solutions. The methanol, 2-propanol, and ethyl acetate meet ACS spacifcstions for analytical reagents. The eth! alcohol ‘was redisiled over potassium hydroxide mado acording to United States Pharmacopeie, and the moths! Cellosive was redistiled from technical grade stock Absorbance of Color Developed with Chromium. Te absorb tance of tho chromium diphenylearbazide color vuried inversely ‘withthe amount of discoloration developed by the solution ofthe reagent (Figures 1 and 2). Upon aging, the aqueous acetone solutions reasted with ehromfum wo give éolors with low absorbs ance, while the slightly acidic nonaqueous, acetone solutions gave colors having consistently high absorbances. ‘The ethyl acetate solutions also gave consistent, although slightly lower, absorb ‘As a rule the freshly made solutions gave the highest absorb- tance values. ‘There was a tendency for the aqueous and ethyl ‘loohol solutions to drop relatively sharply i the Stet 1 or 2 days, Iovel off for 1 to 2 weoks, and then gradually taper off to lower anil lower color intensities, Tt is advisable to standardize SEEN ae “necro, ace. s08 40 1388 all solutions carefully in terms of time, as some lose sensitivity even within the first few hours, Once sensitivity. has ‘ueen lost it cannot be regsined even with exrass reagent (4). susMARY ‘The stability of solutions of eiphenyl- carbaride, colorimetric reagent for chromium, ‘ie greater in nonaqueous ‘organic solvents than itis in the solue tions eonteining water. As the solu- tions of the reagent age, they tend to we ‘become yellow-brown to orange-red and the sensitivity of the reegent varies inversely with the amount of discolora- tion developed. Once sensitivity: hae tyeen lost it cannot be regained even ‘Therefore solutions which become dis colored should be discarded or reealibrated Eth] agetate, high purity acetone, of acetone meeting ACS specifications which does not have a tendenes to be basi, ean, bbe uted to make solutions which will be etable for months and five chromium diphenslessbaside colors of consistently high Absorbance, Metby! ethyl Ketone, methsl Cellocolve, 2-pr0- ppenol, and other similar types of organi solvents make solutions ‘sable for 1 t0 2 weeks, Th all probability the elitination of traces of water, bssie substances, and oxidizing agente would rake these and similar solutions table for stil Toner periods of time. The use of glacial scetic acid-ethy! sleohol solutions fand phthalic anhydride-ethoT aleobol solutions ie in part agreement with these principles, since these solutions ae acidic fd tend to restrict the chemfesl activity of the water that le present, Refrigeration of the solutions helps, but the refrgenae tion effect it minor compared to the effect of the txpe of solvent sed, ‘Substances tending to he basic, stich as methanol, ethyl ‘Aloohol, and thoce containing traces of water or basic impurities, do not make good solvents for stock sabations of the calorimetti reagent. Aaiicous solutions of the reagent should be avoided, ACKNOWLEDGEMENT Appreciation is expressed to Robert L. Townsend for his faithful technical assistance in gathering the data for this paper. rrenctere cre, (2) Alloa, C.F. HL, “Organi Sythe, ‘York, 1040 @) Bose, din Nalure, 170,218 0 {G) Cazenove, Pa Bull oe shin. France, {3} 28, 70 (190035 3] "25, Ta (1001) ) Davis, H.C. ad Bacon, A. J. Sax Chem. Ind, (London), 6, ‘Ste sis), (6) Be, Je Ped and Silverman, Lestio, Awat. Cust, 19, 698 ‘ion. (@ Feeh ! Sede pp. 128 “qualitative Analynin by Spot 2, Flsowien, New York Teo. Bn. Cities Asta Hips 1 412 (989), fon of Traces of Bete ite" Onde, p. 114, Intersioncs, Now York 1980 (iu) Sandal, EB. Tho, Bye! Cyst, Ava Eo., 8 $35 (090), (02) Silverman. Lelio, aod Bae, 1B Sey J Bnd Hy. Tova, 28, 136 coi). (a9) Urone.P-Fvand Anders H. K., Aut. Cis, 22, 1817 1980). (1) Weleton FF, "Ozganie Anaivioa Reagontn Vel IE. 480, ‘Van Nira, New York, 1047,