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T H E J O U R N A L O F I X D C S T R I A L A N D ENGIi!T~EERI-VGC H E M I S T R Y
probably a much higher occurrence of carbonate t h a n would be found in. virgin or cultivated calcareous soils adapted t o plant growth. I n cases of soils running over 1 5 per cent C a C 0 3 , i t would be well, probably, t o increase t h e strength of acid t o I/:. Two soils treated with a n exceedingly insoluble dolomite t o the amount of chemical equivalence of zoo,ooo pounds of CaO per two million pounds of soil gave t h e following results: P E R CEHTCaCOs in Soil A Boiled, minus boiling blank on original soil. . , . . . 13.69 l / 5 HC1, room temp., 30 min. agitation and aspiration ................................. 13.80
Soil B 14.21
Av. 13.95
14.10
13.95
As emphasized b y italics in Bzilletiiz 100, we were able t o secure b y t h e use of 111: phosphoric acid complete decomposition of a very insoluble dolomite,' when i t was EXCEEDISGLY F I F E . I t would not be feasible, however, t o grind all miscellaneous and unknown soils t o such fineness as would be essential, in order t o insure complete decomposition b y 1 / 1 5 H3P04of a possible exceptional occurrence of dolomite. F o r this reason, together with the substitution of the absorption tower for t h e gravimetric procedure, we modify t h e method b y t h e suggestion t h a t 1/10 HC1 be used for miscellaneous work. Instances mhere considerable residual magnesite might be expected t o occur in soils would be exceedingly rare. We shouid probably note, however, t h a t we have found t h a t in such cases neither the old procedure of boiling for one minute or several minutes, nor t h e Marr method, nor the Tennessee Station method will effect t h e complete decomposition of t h e mineral magnesium carbonate. I n soils treated escessively with magnesite a t this Station, we found it essential t h a t boiling be continued during t h e go-minute period of aspiration in order t o decompose completely all of t h e magnesite. MANIPULATION O F MODIFIED PROCEDURE
Twenty-five cc. of approximately 4 per cent N a O H solution are placed in each absorption tower, and COZfree distilled water added t o cover t h e beads. The suction, which is applied before running t h e acid f r o m t h e bulb of t h e inlet t u b e into the 300 cc. Erlenmeyer soil-container flask, is controlled b y screw corks on. t h e rubber tubing t o better advantage t h a n by use ,3f the metal stopcocks of t h e suction system. After agitation and aspiration for 30 minutes, t h e caustic i s drawn o u t of t h e towers into glass tumblers and t h e towers are completely filled with Con-free water. After this washing has been drawn t h e tower should be rinsed several times. T h e N a O H solution is t h e n t i t r a t e d almost t o a disappearance of t h e color given by one cc. of phenolphthalein, by t h e use of approximately normal sulfuric or nitric acid and t h e red color is t h e n completely removed with t h e standardized weaker acids, N / 2 a n d N/2o. TWOdrops of methyl orange are t h e n added and t h e COZ is determined b y titration t o t h e end point of methyl orange. For t h e average soil t h e entire methyl orange titration is carTied o u t by t h e use of A T / 2 0 acid, b u t with excessive coZoccurrences, i t is convenient t o use N / 2 acid t o t h e near end point, and t h e titration is then completed a+th N / 2 0 acid, which has a C o n value of 0 . 0 0 2 2 g.
Vol. 7 ,
KO.3
per cc. In determining t h e blank on t h e S a O H w e also include t h e atmosphere blank of the apparatus. This atmosphere blank is usually exceedingly small and constant, and its use eliminates t h e necessity of sweeping t h e apparatus free of C o n before making determinations. For t h e average soil 20-gram charges should be used and 60 t o 7 5 cc. of 1 / 1 0 acid added for decomposition of t h e carbonates. A nitrogen bulb should be inserted as a t r a p between t h e bottle containing S a O H for purification of incoming air and t h e acid bulbs of t h e apparatus, A CORRECTION
As above stated, i t was'found t h a t i t is necessary t o have dolomite in t h e form of a floury powder in order t o insure its complete disintegration b y 1 / 1 5 phosphoric acid and in t h e original C o n determinations upon t h e dolomite used for treatments, t h e analyses of which were given in Table 1 7 , Tennessee Station, Bull. 107, the analyzed samples were so conditioned. However, in t h e supplementary determination for residual CO? in t h e field samples reported in Table V I of t h a t bulletin the soil samples were ground t o pass a IOO mesh sieve. Upon recently submitting t h e composites of these t w o sets of eight soils each t o boiling and applying t h e boiling blank of t h e untreated soil, 0 ~ 4 4 1per cent of COz was found in t h e limestone residues while 0.45o;per cent was obtained from residual dolomite. T h e same composite soils, when analyzed by t h e use of 1/10 HC1 a t room temperature, gave 0.481 and 0 ~ 4 9 0per cent of residual COP for limestone and dolomite treatments, respectively. This means t h a t the results for residual carbonates of dolomite treatment given in Table VI of Bulletiiz 107 are erroneous, in t h a t t h e y are too low as a result of insufficient fineness of'samples. All of t h e other tables of t h e residual carbonates, however, were verified b y boiling and applying t h e boiling blank prior t o t h e publication of t h e data. T h e writers would qualify their previous conclusions, then, as t o t h e complete disappearance of the magnesium carbonate of t h e C O A R S E L Y G R O U N D dolomite in t h e short length of time given. The general principle, however, is unaltered, as relating t o t h e unusual affinity of magnesium carbonate for siliceous substances. And the corrected results for the residual carbonates indicated t h a t the greater affinity between magnesium carbonate and silicates has offset t o a n extent the lesser solubility of the dolomite in carbonated water, a s compared t o t h e greater solubility of limestone. DEPARTMENT O F CHEMISTRY A N D AGRONQMY UNIVERSITYOF TENNESSEE EXPERIMENT STATION KNOXVILLE _.
THE ,CHEMISTRY OF BASE GOODS FERTILIZER By ELBERT C. LATHROP Redeived Kovember 16, 1914
I ' N T R OD U CTXO N
Although i t has been long recognized t h a t organic matter is a necessity for soil fertility, i t is b u t lately t h a t t h e deeper sigaificance of soil organic matter in
M a r . , 1915
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T H E J O U R N A L OF I N D U S T R I A L A N D ENGINEERING C H E M I S T R Y
relation t o t h e complex problems of t h e soil a n d of t h e crop production has become apparent. Investigations have shown t h a t t h e organic compounds i n soils n o t only affect t h e physical conditions a n d chemical reactions of t h e soil b u t t h a t t h e y are also directly connected with fertility or infertility, some of t h e m being essentially beneficial t o t h e growth of plants while others are distinctly harmful. Of t h e organic compounds t h u s far isolated from soils, a large number contain nitrogen, a n d of these nitrogenous substances, some have been found rather widely distributed in soils varying as t o location, climate, methods of cropping, etc. These nitrogenous compounds either occur a s plant constituents or arise from t h e decomposition of plant or animal protein, brought a b o u t b y t h e various biological a n d bioch’emical agents i n t h e soil. N o t only compounds of t h i s class found i n soils b u t also m a n y other protein decomposition products have been studied, both alone a n d in conjunction with t h e three fertilizer elements, in respect t o their action on plant growth, a n d t h e y have been shown in a number of cases t o exert a beneficial influence; furthermore, these complex compounds are available for use b y t h e plant without first being changed b y chemical or biochemical means into ammonia a n d t h e n t o nitrates.’ T h a t these facts have a n immense practical bearing on fertilizers a n d t h e fertilizer industry, both from t h e standp6int of t h e producer and of t h e consumer, is a t once obvious. The consumer of commercial organic fertilizers should be concerned not alone with t h e availability of t h e nitrogen in fertilizer b u t also in knowing t h e nature of t h e organic compounds he is adding t o his soil a n d in understanding t h e physiological effect of these on plant growth, whether their action will be harmful or beneficial or a balance of these two effects. A chemical investigation of some of t h e types ,of commercial organic fertilizers with t h e view of determining t h e kinds a n d nature of t h e organic compounds in t h e fertilizer is therefore of interest t o t h e agriculturist. T h e old high-grade nitrogenous fertilizers, such as cottonseed meal, dried blood, fish scrap, etc., are being used more and more for feed purposes, a n d t h e time cannot be far distant when their use as fertilizers will cease t o be economic; t h u s a necessity for other a n d cheaper fertilizers of this t y p e arises. On t h e market are a large number of fertilizers which may be characterized as “processed,” i. e., t h e crude materials, not in themselves permissible as fertilizers, are made t o undergo some decided chemical change t o render t h e m suitable a s plant nutrients. I t has been found t h a t t h e “availability” of t h e crude substances is nearly always greatly increased b y such processing a n d t h a t a much larger percentage of t h e nitrogen i n t h e finished product is soluble in water, although t h e actual chemical changes produced seem t o have received little a t t e n tion. T h e chemical compounds i n process fertilizers which are here shown t o have direct fertilizer 1 “A Beneficial Organic Constituent of Soils. Creatinine,” Oswald Schreiner, E. C. Shorey, M X Sullivan and J. J. Skinner, Bull. 83, Bur. Soils, U S Dept. Agr., 1911. “Nitrogenous Soil Constituents and their Bearing on Soil Fertility,” Oswald Schreiner and J. J. Skinner, Bull. 87, Bureau of Soils, U. S. Dept. Agr.. 1912.
229
significance have not been determined, other t h a n t o show t h a t ammonia is formed during processing a n d t h a t ammonia is more readily produced from t h e processed goods. Since t h e wastes from which t h i s t y p e of fertilizer is made contain more or less protein, or protein-like substances, i t seemed quite obvious t h a t t h e finished fertilizers must contain more or less of t h e chemical compounds which would arise from pure proteins b y similar t r e a t m e n t in t h e laboratory. As t h e action on plants of rnany of this class of compounds has been determined i t is evidcnt t h a t t h e finding oE such compounds in t h e fertilizers would throw much light on t h e question of t h e “availability” of t h e nitrogen in t h e fertilizer itself. BASE G O O D S A TYPE O F PROCESSED PERTILIZER
For a chemical s t u d y of processed fertilizers a sample of “wet-mixed” or “base goods” fertilizer was chosen as a representative of this t y p e of fertilizer material. T h e base goods was obtained directly from t h e factory for use in $his investigation. This fertilizer is made b y t h e t r e a t m e n t of various t r a d e wastes a n d refuse, such as hair, garbage tankage, leather scraps, etc., with rock phosphate a n d t h e requisite a m o u n t of sulfuric acid. These materials are mixed together in a “ d e n ” a n d t h e resulting mass is allowed t o s t a n d for several days, or until i t is cool enough t o be conveniently handled. I n t h e course of t h e reaction t h e mass reaches a temperature approximating 100’ C., a n d t h e identities of t h e original substances are almost or entirely lost. Under these conditions it is certain t h a t ’ m o r e or less hydrolysis of t h e proteins in t h e crude materials takes place, with t h e formation of proteoses, peptones, polypeptides, or t h e simple amino acids, t h e kinds a n d number of products formed necessarily depending on t h e proportion of t h e different proteins in t h e original materials, o n t h e amount a n d strength of t h e acid, t h e length of time of t h e reaction, a n d t h e temperature reached during t h e treatment. Hartwell a n d Pember’ have recently made a s t u d y of base goods in order t o determine t h e availability TABLEI-PERCENTAGETOTAL KITROCEN I N CRUDEMATERIALS AND FINISHED PRODUCT (HARTWELL AND PEMBER) Hairtankage.. , .. 6 . 2 8 B Roasted leather. . , 6 . 4 9 WaterGarbage tankage., , . 2 . 8 7 Water-
.
TABLE 11-PERCENTAGEOF
THE TOTAL NITROGBN PRESENTIN DIFFERENT FORMS(HARTWELL AND PEMBER)
. . . . . . . . . .. .. .. .. ..
In ammonia.. . . . . . , . . . . . .. . In water-soluble organic matter.. In water-insoluble organic matter.. , . ,
Before After put removing into den from den 14.3 6.5 57.7 7.8 28 .O 85.7
of t h e nitrogen contained compared with t h a t of t h e high-grade nitrogenous fertilizers. T h e product t h e y used was made from hair tankage, garbage tankage, a n d roasted leather, together with rock phosphate a n d sulfuric acid. Tables I a n d I1 give their figures for t h e analysis of t h e crude materials used i n producing t h e fertilizer a n d of t h e finished product. T h e experimental work of t h e present investigation was along t w o separate lines: THIS
JOURNAL, 4 (1912), 441.
T H E J O l i R N A L O F I N D U S T R I A L A N D ENGIATEERING C H E M I S T R Y
230
I-Analytical, involving total nitrogen determinations and t h e se,parate estimation df t h e various forms in which nitrogen m a y occur. 11--A determination of the definite chemical compounds present in t h e fertilizer b y suitable methods of isolation and identification. THE CHEMICAL CHANGES IKVOLVED I N PROCESSIXG
It is not t h e purpose of t h e present paper t o enter in detail into t h e methods used in t h e chemical examination of t h e base goods. For t h e analytical study of t h e forms of nitrogen in t h e base goods t h e method of Van Slyke' was used in its essential details, omitting t h e determination of cystine, since t h e method for its determination is n o t applicable in this case. It should also be stated t h a t , although t h e results from t h e Van Slyke analysis are expressed in t h e usual way, arginine nitrogen, histidine nitrogen, etc., i t is not intended t o convey t h e impression t h a t these fractions contain pure arginine, histidine, etc., since, as will be shown later, other compounds are included under these analytical terms. However, t h e nitrogen so expressed is t h a t contained in compounds which give t h e various reactions upon which the Van Slyke method depends. T h e analytical figures obtained b y this method appear in Table I V and will be discussed after t h e isolation of certain of t h e constituents has been presented. The method used for isolating and identifying t h e various organic compounds are such as have been used in this laboratory in separating these compounds from soils and are described in various publications from this l a b o r a t ~ r y . ~ Since this investigation aims only a t an explanation and exposition of t h e general chemical principles involved in the treatment of trade wastes and other organic material t o render t h e nitrogen contained therein more available for agricultural purposes, i t does not aim t o present t h e research methods here employed as general methods for analyzing such fertilizers. Nor can t h e quantitative figures obtained be expected t o apply t o all products of similar manufacture, for t h e reason t h a t t h e different kinds of nitrogen compounds will necessarily show different proportions according t o t h e nature of t h e materials which enter into t h e mixture. T h e compounds which mere isolated from the base goods are tabulated in Table 111 according t o the sources from which they have been derived and t h e chemical groups t o which they belong. TABLE 111-ORGAXIC C O X P O U N D S ISOLATED F R O M SAlrlPLE OF RASE GOODS Arginiue. . . . . . . Diarnino acids or Products of protein hydrolysis by Histidine.. . . . . . hexone bases acid treatment of raw materials. Lysine.. . . . . . . . Leucine. . . ,. ,.,.,.,. Monoamino acids Guanine . . . . . . . Purine base Plant constituent, or product of hydrolysis of nucleoprotein. Hypoxanthine. , Purine base Plant constituent, or product of conversion of nucleoprotein-base.
}
ryrosine,, 1
}
Establishing t h e presence of these prod.ucts of acid hydrolysis of proteins, namely, t h e diamino acids, J. Biol. Chem., 10 (1911), 15-55. Oswald Schreiner and E. C. Shorey, Bur. Soils, 72, S. Dept. Agr., 1910; Oswald Schreiner and E. C. Lathrop, Bull. 80, Bur. of Soils, U. S. Dept. Agr., 1911; E. C. Shorey, Bull. 88, Bur. Soils, U. S. Dept. Agr., 1913; Oswald Schreiner and E. C. Lathrop. BuEl. 89, Bur. Soils, U. S.Dept. Agr.. 1912; E. C. Lathrop, Bull. 158, U. S . Dept. Agr., 1914. 1
2
Vol. 7, No, 3
arginine, lysine, a n d histidine, and t h e t w o monoamino acids, leucine and tyrosine, in the amounts in which t h e y were found is of itself sufficient evidence t o demonstrate t h a t b y t h e acid treatment of t h e crude materials used in t h e manufacture of t h e base goods t h e proteins contained therein have been changed. T h e change is shown t o be a deep-seated one, since five of t h e compounds known t o be final products of protein hydrolysis b y acids are found. This, however, cannot be t a k e n t o mean t h a t t h e proteins have been completely hydrolyzed b y t h e acid treatment since i t is possible t o have present in t h e product of partial hydrolysis of proteins, not only t h e diamino a n d monoamino acids, b u t also such intermediate compounds a s polypeptids, peptones, proteoses, etc, I n this connection t h e results obtained b y use of t h e Van Slyke method, and given in Table IV, are of particular interest. For t h e nitrogen partition two samples of base goods were extracted: ( I ) with boiling water and ( 2 ) with boiling acid. I n t h e former case only slight further hydrolysis of the materials in the base goods is t o be expected since t h e free acid in t h e fertilizer is extremely weak, and t h e boiling temperature, zooo C., is t h a t which was reached in t h e process of manufacture. I n t h e case of t h e second extract complete hydrolysis of all. t h e proteins or protein-like materials is certainly t o be expected, since In addition t o t h e original hydrolysis t h e material was boiled with strong hydrochloric acid for 2 4 hours, a treatment sufficient for t h e complete hydrolysis of most proteins. T h e differences in the results obtained from t h e analyses of t h e two extracts may, therefore, be expected. t o throw some light on t h e question of t h e completeness of hydrolysis of the original proteins b y t h e acid processing, TABLE IV-KITROGEN FORXS AS
c1ETERMIKED BY THE V A X S L Y K E METIIOD
Results expressed in per cent of
EXTRACT
T o t a l nitrogen.. . . . . . . . . . . . . . . . T o t a l soluble nitrogen.. . . . . . . . . . T o t a l insoluble nitrogen.. . . . . . . . Amide nitrogen.. . . . . . . . . . . . . . . Humin nitrogen., . . . . . . . . . . . . . . D i y r ~ o Arginine N . , . . . . . . . Histidine PIT.. . . . . . . . fraction Lysine N . . . . . . . . . . Monoamino acid Amino N . . , , fraction Konamino N , . (a) Obtained indirectly.
{
1
Base goods
Total N in base goods
HzO HCI HZO €IC1 1.610 1.610 85 i 4 ( a ) a i : i 4 1.372(a) 1.435 0.238(a) 0. 175(u) 14. 76(a) I I .36:a) 0.374 0.382 23.23 23.70 0.074 0.031 4.61 1.95 6.46 0.111 0.104 6.89 0.117 0.070 7.26 4.38 0.117 7.26 0.081 5.06 0.546 3 3 , 75 0.543 33.92 7.10 0,133 8.27 0.114
:
Without discussing t h e significance of all t h e figures obtained by t h e use of this m e t h o d i t is interesting t o note t h a t t h e differences betn-een the water extract and hydrochloric acid extract indicate t h e presence in t h e base goods of a nitrogenous compound not yet completely hydrolyzed. I n order t o prove t h e presence of some intermediate product of protein hydrolysis, which is t h u s indicated b y analytical methods, an aqueous solution of about 2 . 5 lbs. of base goods was made and t h e diamino acids mere precipitated with phosphotungstic acid in t h e presence of 5 per cent sulfuric acid. T h e precipit a t e which formed was allowed t o stand over night and after filtering OH it was mashed'well with j per cent sulfuric acid and dissolred in sodium hydroxide. T h e phosphotungstic acid was then precipitatcd by adding barium hydroxide sol.ution, and after filtering, t h e excess of barium mas removed b y adding sulfuric
I
Mar.,
IQIj
T H E J O U R N A L OF I N D U S T R I A L A N D ENGINEERING C H E M I S T R Y
2.31
acid until a neutral reaction was obtained. Portions trade’ wastes, such as hair, lather, garbage, etc., is of t h i s solution were tested for peptones, proteoses, made more available, is recognized as a process of etc., with t h e following results: ( I ) T h e biuret test partial hydrolysis of t h e complex protein contained was positive; ( 2 ) a precipitate was obtained on satura- in such materials, resulting i n ammonia, amino acids, tion of t h e solution with ammonium sulfate, or with etc., all of which are more available t h a n t h e original TABLE \’-PRODUCTSOF ACID HYDROLYSIS OF VARIOUSPUOTEINS sodium chloride; (3) when t h e filtrate from t h e l a t t e r “Synotin” “Keratin” from “Legsolution was treated with acetic acid a cloudy prefrom Haliumm” cattle sheep’s sheep’s horse’s b u t Ox from cipitate developed; (4) precipitates were also obflesh horn wool hair muscle muscle pea COMPOUND (ai (bl (bl (c) ie) it3 ., ., . , (,d,i ., ~.,, tained with sulfuric acid, hydrochloric acid, phosGlycine (9) , . . . . , . . . . 0.5 0.5 0.6 0.0 2.1 0.4 4.7 phomolybdic acid a n d with phosphotungstic acid; (5) Alanine(g). ...... . . . . 4 . 0 1.6 4.4 3.7 2.1 1 . 5 (?) . . . . . . . . . . . . . . 0.9 4.5 2.8 0.9 0.8 0.8 a precipitate was formed in t h e addition of alcohol Valine Leucine(g) . . . . . . . . . . . 7.8 15.3 11.5 7 . 1 10.4 11.7 8:O Isoleucine.. . . . . . . . . . . . . . ... t o t h e solution. This precipitate was filtered off, Phenylalanine(g) . . . . . 2 . 5 1 9 0:O 3:l 3.2 3:8 Tyyosine(g).. . . . . , . . . 2.2 3.6 2:9 3.2 2.4 2.2 1.6 dissolved i n dilute alkali, a n d on addition of very dilute Serine ......... . . . . . . . . 1.1 0.1 0 . 6 (?l (?i 0.5 .. .. 7.5 7.3 8.0 ,,, ,,,,,... copper sulfate solution t h e biuret reaction was again Cystine, Proline . , . , . . . , . . . . . . 3:s 3.7 4.4 3.4 i:2 5:s 3 : ~ obtained. These reactions are those which are given Oxyproline, . . , , , , . . . Asparticacid(g) . . . . . . 0 : 5 2:s 2:3 0:3 218 4:52 5 : 3 by proteoses a n d b y t h e proteins a n d confirm t h e con- Glutamic acid(g). , . . . 1 3 . 6 1 7 . 2 1 2 . 9 3 . 7 1 0 . 1 15.5 1 7 . 0 ., , .. .. clusions arrived a t from t h e results with t h e Van Slyke Tryptophane. (69 /l? Arginine(g) . . . . , , , . . . 5 :1 2 7 . . 2:5 .. . .. .. , . . . 3 . 3 0.2 .. .. 1.1 7.5 7.6 5.0 method. T h e Millon reaction a n d t h e Hopkins-Cole Lysipe,(g). Histidine(al . . . . . . . . . 2.7 .. 0.6 2.6 1.8 1.7 Ammoniacg). . . . , . . . . 0 . 9 . . . 1 . 4 1 . 1 2.1 reaction were both negative, showing t h e absence 47.3 6 2 . 3 49.2 39.6 5 0 . 7 67.5 6 2 . 4 from t h i s protein-like compound of t h e tyrosine a n d (a) Abderhalden and T. Saski Z.physiol. Chem. 5 1 (1907) 404. t h e t r y p t o p h a n e radicals. ( b ) E. Abderhalden a n d A. VAitinovici, Ibid., 6a (1907), 34‘8. Abderhalden a n d H. G. Wells, I b i d . , 46 (1905), 31; A. Argiris. A very large number of compounds intermediary Ibid.,(c)54 E.1905), 86. (d) B. Osborne a n d F. W Heyl Am. J . Physiol. 22 (1908), 433. between t h e protein and its primary hydrolysis prod( e ) T. B. Osborne a n d D. B: Jonei, I b i d , , 24 (1909): 437. ( f ) T.B. Osborne and F. W. Heyl, J . Bzol. Chem., 6 (1908), 197. ucts m a y occur, depending on a great variety oE con(g) Physiological action on plant growth has been determined a n d reported in Bull. 87, Bureau of Soils, U. S . Dept. Agr. ditions, so t h a t t h e actual identification of t h e compounds under discussion would be a difficult matter. protein material; t h i s hydrolysis is almost complete, However, t h e n a t u r e of this compound m a y be approx- t h e nitrogenous compounds formed being principally imately determined b y t h e results obtained in t h e t h e primary products of protein hydrolysis, together s t u d y of t h e t w o extracts b y t h e Van Slyke method. with a small a m o u n t of proteose-like compound which These results indicate t h e presence in t h e base goods has not been fully decomposed. of a compound of a proteose nature, which, since it A V A I L A B I L I T Y O F T H E N I T R O G E N O F O R G A X I C P E R gives a biuret test, must be composed of a t least TILIZERS three amino acids. The results indicate still further The question of t h e availability of t h e different t h a t t h e compound is composed of acid amide radicals, kinds of nitrogen contained in organic fertilizers is diamino acids-particularly lysine, a n d monoamino one t h a t has caused considerable discussion. A n u m acids-those containing amino nitrogen a n d espe- ber of methods have been proposed for determining cially those containing non-amino nitrogen. Since t h e this factor, a n d while some of t h e m give helpful refigures obtained b y t h e nitrogen partition method are sults, all except t h e plant method are open t o more or subject t o a certain amount of error when applied t o less objection, since t h e methods are empirical a n d such a mixture t h e figures can be taken as only a p - t h e nature of t h e complicated compounds i n which proximate for t h e various forms of nitrogen which t h e nitrogen is linked in t h e fertilizer is unknown make u p t h i s compound. or only guessed. When these nitrogen compounds I n Table V are given t h e primary hydrolysis prod- are known a n d the’u action on plants, as well as t h e ucts of a number of proteins which may be present action of t h e compounds formed from t h e m during i n t h e base goods. These results were obtained b y their decomposition in t h e soil, have been determined, t h e esterification method a n d show how t h e different t h e n t h e question of t h e availability of t h e nitrogen proteins vary i n t h e n a t u r e a n d a m o u n t of t h e units of organic fertilizers can be understood. Originally composing them. M a n y monoamino acids, besides it was held t h a t plants were able t o use nitrogen only leucine a n d tyrosine, occur in these proteins, a n d there when i t was offered t o t h e m i n t h e form of nitrates; must consequently be present in t h e base goods amino this idea, however, was modified when it was discovacids other t h a n t h e two isolated. This is apparent ered t h a t under certain conditions plants used ammonia from t h e composition of t h e various proteins shown in or ammonium salts without their conversion i n t o t h e table. Owing t o t h e large a m o u n t of amide nitrates quite as well as t h e y used t h e nitrates t h e m nitrogen present in t h e fertilizer, which was split off selves. During t h e past few years it has been clearly b y t h e acidulation of t h e original proteins of t h e t r a d e demonstrated t h a t plants use nitrogen n o t only in t h e wastes, it m a y be concluded t h a t considerable quanti- form of nitrates a n d ammonia b u t in t h e form of comties of aspartic or glutamic acids are present in t h i s plex organic compounds.’ T h e action of a number sample of base goods. of these nitrogenous compounds has been tested in The conclusion t o be drawn from t h e results ob- this laboratory in conjunction with t h e three fertilizer tained b y t h e examination of this fertilizer b y means elements a n d it has been found t h a t in some cases of t h e analytical a n d isolation methods a r e as fol1 Hutchinson a n d Miller, Centv. Bakl. Parasilenk, 80 (1911), 513; lows: T h e process b y which t h e nitrogen of certain Schreiner a n d Skinner, Bull. 87, Bureau of Soils, U. S. Dept. Agr., 1912, A -
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a32
T H E J O U R N A L O F I N D U S T R I A L A N D ENGIATEERIAVG C H E M I S T R Y
t h e nitrogen compounds are not only used as a source of nitrogen for t h e growing plant, without a n y change in t h e compound, b u t t h a t they are apparently nitrate sparers, i. e., t h e plant uses t h e m in preference t o t h e nitrates. Instead, then, of only one kind of nitrogen compound, ??ifrate,or a t most two, izitrate and ammonia, there appear t o be a very large number of nitrogenous compounds which have properties of physiological importance t o plant growth. T h e question of t h e availability of nitrogen compounds can therefore be answered only when t h e nitrogen compounds contained in t h e fertilizer can be determined in amount and a t t h e same time classified according t o their physiological action on plant growth. I t is hardly necessary t o s t a t e t h a t such a method does not exist a t present a n d t h a t t h e physiological action of only a p a r t of t h e total number of nitrogenous compounds present in fertilizers is known The physiological action on plants of all of the nitrogenous compounds isolated from base goods has been determined by means of water cultures' a n d t h e results obtained may be stated briefly, as follows: Both of t h e purine bases are used b y t h e plant as a source of nitrogen a n d are beneficial t o plant growth; furthermore, t h e hypoxanthine acts as a nitrate sparer, there being less nitrate used b y t h e plant in t h e presence of hypoxanthine t h a n when t h e hypoxanthine is absent. Histidine, arginine, and lysine2 are all beneficial t o plant growth, causing nitrogen increases in t h e plant, and t h e two first diamino acids act as nitrate sparers; this may also be t r u e of lysine, although this property of lysine has not been studied. Leucine is also beneficial t o plant growth, b u t tyrosine, in t h e light of later investigations, is somewhat doubtful in action. Of t h e other monoamino acids which may be present in base goods, aspartic acid, glutamic acid, and glycocoll have been found t o be beneficial. T h e action of alanine is somewhat doubtful, i t apparently being beneficial in low concentrations, while phenylalanine is reported as harmful. T h u s six of t h e seven compounds isolated f r o m t h e base goods are actually available t o plants as such a n d have a beneficial action. Of t h e monoamino acids, other t h a n t h e two isolated from base goods, which have been studied in regard t o their action on plant growth, three have been found t o be beneficial, one doubtful, and one is reported as being harmful. The high-grade nitrogenous fertilizers. such as dried blood, are considered t o have a high availability ovtling t o t h e fact t h a t t h e nitrogenous materials when placed in t h e soil quickly undergo t h e process of ammonification and nitrification, t h e nitrogen t h u s being changed into a form which can be immediately used b y the plant. I n fact, Lipman3 has proposed a method f o r t h e determination of t h e availability of t h e nitrogen of organic fertilizers, depending on t h e amount of ammonia produced under certain conditions in a gi17en length of time. It is evident from t h e above Con'sideration t h a t such a method does not tell t h e whole 1
Bull. 87, Bureau ot Soils.
* Unpublished data.
s Lipman, Brown and Omen, Centr. Bakt. (191 1-1912). 49-85.
Parasitenk, 31, II A b t .
Vol. 7 , No. 3
story, since, in the decomposition of protein materials, such as dried blood, intermediate compounds are formed which are undoubtedly in themselves beneficial t o plant growth. In order, therefore, t o understand t h e complete action of the nitrogenous materials in t h e base goods i t is necessary t o know how t h e compounds contained in i t are acted upon b y ammonifying bacteria. Jodidil has shown t h a t t h e amino acids a n d acid amides are quite readily ammonified when placed in t h e soil, t h e rate of ammonia formation and t h e amount of ammonia formed depending apparently upon t h e chemical structure of the particular compound under consideration. In general, he found t h a t t h e simpler t h e chemical structure of t h e nitrogen compound t h e more quickly and readily it was ammonified. In t h e light of these Eacts i t appears t h a t polypeptides, peptones, proteoses, and proteins would be ammonified still more slowly t h a n t h e amino acids since their structure is increasingly more complex. Hartwell and Pember2 in their study on t h e availability of t h e nitrogen of base goods, b y means of plant tests found t h a t i t h a d apparently as high an availability as dried blood, t h e water-soluble nitrogen having even a higher availability. From t h e nature and amounts of t h e compounds present in the base goods this might be predicted. I n t h e case of t h e dried blood, t h e nitrogen is practically all in t h e form of complex protein material which must be broken down into simpler compounds b y bacterial action, with t h e formation of ammonia and other nitrogenous compounds, some or all of which may be of physiological importance t o plants. With t h e base goods t h e case is a little different; the greater part of t h e nitrogen is a t once available for plant use, and a t the same time these available compounds may be changed more easily and quickly b y bacteria of t h e soil into ammonia and nitrate, which in t u r n are used b y t h e plant. The soluble nitrogen of base goods should therefore be in a more readily a\Tailable form t h a n the nitrogen of dried blood or other nitrogenous fertilizers which are entirely of a protein nature. THE CHEMICAL P R I K C I P L E S U K D E R L Y I N G
T H E L-TILIZA-
T I O N O F N I T R O G E N O U S T R A D E WASTES
It has already been stated t h a t in order t h a t the plant may make use of t h e nitrogen of even highgrade organic fertilizers, i t is necessary for t h e proteins therein t o be a t least partially decomposable b y t h e biological and biochemical agencies of the soil. T h e low-grade organic nitrogenous fertilizers resist decomposition b y these biological and biochemical soil agencies, and their nitrogen is therefore considered t o be less available for plant use. The guiding idea behind the processes proposed for t h e treatment of trade wastes, which will not decompose easily in the soil as such, is t o change t h e nitrogen compounds contained in them in such a way t h a t ammonia is formed and t h a t their decay in t h e soil is more rapid. Much of t h e nitrogenous materials in trade wastes is of a protein nature, since t h e products from which 1 2
Research Bull. No. 9 , Iowa Expt. Sta. Eoc. cit.
”rrir”,
FIG. ~ - T A N K SUSEDFOR STEEPING BARLEYSHOWING AIR LINES
offensive or disagreeable odors must also be avoided. The species of bacteria, moulds a n d wild yeast encountered in air are very many. b u t fortunately only five species of bacteria are dangerous i n a brewery, i. e . , acetic, lactic, butyric, sarcinae or pediocci, a n d Bacillus viscosus. Practically all others. including all t h e common pathogenic bacteria, will not survive fermentation. This is very important from a sanitary standpoint. I n our plant we use air of different degrees of sterility,
FIG.11-PERMANGANATE TANKS
depending upon t h e use t o which t h e air is t o be a p plied. All air used is drawn through a pipe line from t h e top of our highest building b y means of a p u m p which compresses i t a n d conveys i t t o t h e various points where i t is needed.
r1e
__-
6l.b
”-_.
a n d removed b y a n overfloh device. T h e air used in this process is purified by passing i t through a series of large drums filled with a solution of potassium permanganate. As t h e air is sprayed through these t a n k s of permanganate in a very fine spray t h e permanganate not only washes t h e air b u t oxidizes or removes a n y objectionable odors a n d also removes t h e greater part of t h e dust, d i r t , a n d bacteria. After ‘being steeped for a certain length of time, t h e wet barley is conveyed t o a series of compartments kept a t a uniform temperature in order t o allow t h e barley t o grow. During this growth certain changes t a k e place i n t h e barley kernel, various enzymes, such as amylase a n d peptase; develop, a n d during t h e growing process large amounts of carbon dioxide a n d other gases are evolved. It is necessary t o remove these gases a n d at t h e same time provide t h e barley with a fresh supply of oxygen. This is accomplished b y means of air. This air is purified b y passing i t through a series of galleries constructed of perforated metal plates. Through t h e center a n d running t h e entire length of these galleries there is a water pipe equipped with a large number of very fine sprays. The air must pass through this fine spray of moisture. After t h e growing process is complete t h e barley or as it is now called, “green malt,” is transported t o kilns a n d dried. It is t h e n cleaned a n d this represents t h e finished malt. The third a n d perhaps t h e most i m p o r t a n t place t h a t air is used is in t h e fermenting department. T h e green malt is mixed with water a n d a n infusion m a d e ; this is finally boiled with hops a n d t h e resulting wort must t h e n be cooled t o t h e proper temperature a t which t h e yeast is t o be added for t h e fermentation by a series of coolers very similar i n construction t o t h e ordinary condensers used in t h e laboratory for distillation purposes. Air is passed through t h e wort when it is still hot. This air functions mainly as an oxidizing agent. T h e specific nature of this oxidation has not been very thoroughly investigated b u t i t plays a very important r6le in t h e brewing process a n d serves t o render certain constituents of t h e hop, commonly termed hop resins, insoluble; i t also has some action on t h e proteins a n d carbohydrates a t this high temperature. After t h e temperature of t h e wort has been considerably reduced, more air is added, n o t a s a n oxidizing agent, b u t for solution b y t h e cold wort. This dissolved air is necessary for t h e respiration a n d growth of t h e yeast. When t h e wort has been cooled a n d t h e yeast added, more air serves merely t o mechanically agitate t h e yeast a n d wort a n d secure a more or less uniform mixture,
___
-,y L a ~ ~ ~ b . , I ~ d n g ( "SGll compounds has been brought about b y t h e partial hydrolysis of t h e proteins contained i n t h e various .trade wastes used i n t h e manufacture of t h e fertilizer. When proteins decompose through n a t u r a l conditions, be t h e y in t h e soil or out of i t , a certain amount of hydrolysis of t h e proteins takes place a n d if t h e decomposition is allowed t o proceed long enough under proper conditions complete hydrolysis will result. The principle involved in making t h e nitrogenous #material in t h e soil available a n d in increasing t h e availability of low-grade nitrogenous materials by factory t r e a t m e n t is therefore t h e same. I n other words, t h e general chemical principle t o be applied in making available t h e nitrogen of low-grade fertilizers, t r a d e wastes, etc., is t h a t of complete or partial hydrolysis b y a n y suitable means of t h e proteins contained in t h e wastes. Partial hydrolysis of proteins m a y be accomplished b y means of heat, boiling, steaming, heating under pressure; both partial a n d complete hydrolysis m a y be obtained b y treating with strong acids or alkalis, either in t h e cold for a long time or heating t o a high temperature, t h e extent of hydrolysis depending on t h e several conditions. An examination of t h e patent literature shows t h a t all of these means have been proposed for t h e t r e a t m e n t of waste materials, although t h e actual chemi'cal principle of hydrolysis involved in t h e t r e a t m e n t is not considered. Also, i n a number of processes in actual use, various of these treatments are practiced, resulting in different degrees of hydrolysis of t h e original proteins. While t h e availability of t h e nitrogen of a fertilizer depends on t h e substances i n which t h e nitrogen is contained, it also depends on t h e extent of hydrolysis of t h e proteins used in t h e manufacture. It may be s t a t e d t h a t in general t h e more extended a n d final t h e hydrolysis t h e more available t h e nitrogen of t h e compounds formed, since, as has been shown, t h e final products of hydrolysis are utilized b y t h e plants as such a n d are at t h e same time more readily changed into ammonia b y bacteria, etc., t h a n are t h e intermediate compounds produced b y partial hydrolysis. *LI"I
I
S UM MA R Y
T h e base goods used as a t y p e of processed fertilizer
w i n g principally t h e products or prirlidry p r o w l 1 uLcomposition, together with a small amount of a proteose-like compound which has persisted. F r o m t h e sample of base goods were isolated t h e following nitrogenous compounds : two purine bases, guanine a n d hypoxanthine; t h e three diamino acids, arginine, histidine, a n d lysine; a n d two monoamino acids, leucine a n d tyrosine. A proteose-like compound was also obtained a n d i t s general n a t u r e established. By means of t h e Van Slyke Method t h e approximate proportions of t h e different forms of nitrogen contained in t h e fertilizer were estimated, a n d t h e extent of t h e hydrolysis of t h e original proteins was determined. It was also shown b y this method t h a t t h e proteose-like compound was composed of acid amide radicals, diamino acid radicals, especially lysine, a n d monoamino acid radicals, particularly t h e monoamino acids which contain non-amino nitrogen. T h e question of t h e availability of nitrogen is discussed, a n d from a consideration of t h e amount a n d t h e physiological action on plants of t h e different forms of nitrogen present i n t h e fertilizer i t is concluded t h a t t h e water-soluble nitrogen of this fertilizer should have a n availability equal t o or greater t h a n t h e nitrogen of dried blood, or other high-grade fertilizers. These results are i n accord with t h e results obtained b y t h e plant method of determining availability. T h e general chemical principle which underlies t h e method for rendering available t h e nitrogen contained i n most t r a d e wastes, which are t o be used as fertilizing materials, is shown t o be either partial or complete hydrolysis of t h e protein of t h e wastes b y a n y suitable means. T h e more complete t h e hydrolysis t h e more available t h e nitrogen in t h e fertilizer becomes, since t h e products of complete hydrolysis of proteins are not only utilized b y t h e plants themselves as nutrients b u t t h e y are more 'easily ammonified when placed in t h e soil t h a n are t h e more complex compounds, such a s peptones, proteoses, a n d t h e proteins themselves. BUREAUOF SOILS
U. S. DEPARTMENT OB AGRICULTURE, WASHINGTON, D. C.
LABORATORY AND PLANT PURIFICATION AND STERILIZATION OF AIR' By S. BORX AND WM. F. CARTHAUS
Received h'ovember 16, 1914
I n considering t h e question of air, we have t a k e n i t up mainly from t h e standpoint of t h e fermentation 1 Presented before the St. Louis Section of the American Chemical Society, June 8, 1914.
industry, b u t t h e methods employed here could easily be extended t o other fields such as t h e ventilation of hospitals, schools, public buildings a n d factories. Air is one of t h e most important factors in t h e brewing process, a n d a s sterile air can be obtained b y \ none of t h e ordinary methods employed for purifying
