Kov., I914
T H E J O U R N A L OF I N D L - S T R I A L A N D ENGIiVEERING C H E M I S T R Y
T h e a m o u n t of moisture in green alfalfa varies considerably. A sample cut during a rain contained 81 per cent moisture while in other samples t h e moist u r e content was as low as 60 per cent. The average of those samples dried in t h e laboratory was nearly 7 j per cent. This of course is much greater t h a n would be found in h a y c u t under ordinary conditions. T h e samples used for these tests contained about 64 per cent water, while a. sample taken a t t h e same time, b u t dried immediately. contained 74 per cent water, showing a loss of I O per cent from handling a n d lying in t h e pile over night. Mowing, raking a n d hauling on a bright d a y would probably show a greater loss. Assuming t h e moisture content of a h a y t o be 7 j per cent when it reaches t h e drier, i t would require t h e evaporation of 6000 pounds of water per t o n of d r y h a y ; 6000 X 23.2/100 = 1390 pounds of coal per ton of dry hay. 4 t $2.60 per t o n , this would a m o u n t t o $1.81 per t o n of h a y for heat. T o this shou!d be added something like 2 j cents per t o n for interest a n d depreciation of plant a n d a b o u t $1.00 per t o n for chopping, power a n d labor, making a t o t a l cost of $3.06 for drying one t o n , d r y weight, of hay. Assuming t h e moisture content t o be 64 per cent, as was found in t h e trials here shown, t h e production of one ton of dry h a y would require t h e evaporation of b u t 3600 pounds of water, using 835 pounds of coal a t a cost of S1.09. Adding t o this $1.25 for interest, power, labor, etc., a t o t a l cost of $2.34 per t o n is shown. Although these cost figures are only approximations, t h e y lead t o some interesting conclusions. .is it was shown t h a t t h e t o t a l loss in t h e value of h a y due t o field curing a n d handling amounts t o from 20 per cent under favorable conditions t o j o per cent under adverse weather conditions, a n d a s t h e calculated cost for artificial drying is from $2.35 t o $3.00 per ton, producing only choice alfalfa of uniform grade, it is a p p a r e n t t h a t artificial drying may be conducted at a profit in districts where t h e drier can be situated near both field a n d source of fuel. T h e alfalfa meal industry has grown t o considerable proportions in t h e last few years a n d t h e demand is for well-cured alfalfa of good color. Several alfalfa millers have expressed t h e opinion t h a t a h a y of such dryness could be ground a t a b o u t half t h e cost of t h e average hay, t a k e n from t h e stack. Some millers subject their h a y t o a short, prelimi n a r y drying before grinding. A drier r u n in connection with a n alfalfa-grinding plant would be a n excellent combination. This same principle of drying could doubtless be applied t o other fodders a n d perhaps even t o small fruits a n d other f a r m products which are occasionally damaged b y unfavorable weather during t h e drying season. Among t h e other fodders t o which drying might be applicable might be mentioned t h e pea vines from t h e canning factories a n d t h e common weed, sweet clover or yellow Mellilot. Pea vines are often made i n t o ensilage b u t a s such t h e y have b u t a limited demand a n d t h e canner often has trouble disposing of t h e enormous q u a n t i t y of vines which accumulate a n d become a nuisance. Pea-vine meal has been
919
used t o a certain extent as a n addition t o alfalfa meal. Young sweet-clover is used as forage t o a small ext e n t in Europe b u t sun curing does not produce a good hay. Drying in a n oven a t 5 0 ° , however, gave a h a y of good appearance a n d flavor. Samples of these plants examined gave analyses as follows: Protein Per cent
. . . .. ,. .. .. .. .. .. .. .. .. ..
Pea vines . . . . Sweet clover . . . .
17.73 18 85
Diastatic activity 0.54 0.03
Water solubility Per cent 44.2 32.7
T h e moisture contents of t h e two plants in t h e green s t a t e were 84.7 per cent and 83.2 per cent a n d this would probably be t h e principal drawback in curing t h e m for feed. SUMMARY
I-The diastatic activity of alfalfa is greater in t h e morning or after a period of darkness t h a n after a period o f , exposure t o light. There is much more diastase present in t h e plant during t h e warm, active, growth-producing summer t h a n during t h e spring or fall. Young plants contain more diastase t h a n older ones. 11-Drying a t elevated temperatures in a humid atmosphere decreases t h e diastatic activity, even though t h e temperature is only joo. Drying in a current of air with gradually increasing temperature, on t h e other hand, increases t h e activity markedly. Light a n d weathering in t h e field t e n d t o destroy t h e diastase. Rain during curing is very detrimental. 111-Highly diastatic alfalfas generally show a greater solubility in water b y a u t o digestion t h a n samples low in diastase. However, t h e degree of solubility in water cannot be increased above a certain limit. 1x7-The loss in digestible constituents during handling a n d curing in t h e field may v a r y from 2 0 per cent under favorable conditions t o as much as 5 0 per cent under adverse weather conditions. V-Curing by artificial heat, using t h e principle of counter-currents, gives a h a y of better color, odor a n d flavor t h a n can be produced b y other means. The h a y appears t o retain many of t h e valuable properties of t h e green plant, which are ordinarily lost in curing. VI-The cost of artificial drying is estimated t o be less t h a n t h e losses generally sustained in field curing a n d , therefore, it ought t o be possible t o conduct drying a t a profit when t h e drier can be located near both field a n d source of fuel. T h e writer desires t o acknowledge his indebtedness t o t h e late Robert Kennedy Duncan for helpful direction a n d valuable advice in connection with this work. DEPARTMENT OF INDGSTRIAL RESEARCH UNIVERSITY O F KANSAS,LAWRENCE
THE ORIGIN OF VANILLIN IN SOILS-VANILLIN WHEAT AND IN THE WATER IN WHICH WHEAT SEEDLINGS HAVE GROWN
IN
By M. X. SULLIVAN Received August 19, 1914
Vanillin, t h e chief odorous principle of t h e vanilla pod, t h e fruit of vanilla planifolia, has been reported as present in a number of plants a n d plant products. T h e presence of vanillin in plants a n d its possible
920
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
passage into t h e medium of growth is of great interest since ( I ) vanillin has been found injurious t o plant growth1 a n d ( 2 ) it has been found in a number of soils.2 As yet it has not been reported as formed b y microorganisms. Among t h e plants and plant material in which vanillin has been reported are asparagus sprouts, seeds of white lupine, ilex leaves, dahlia tubers, grain of oats, a n d roots of couch grass, potato, pine wood, peat, raw beet sugar, a n d in resins such as Asafoetida a n d Siam b e n ~ o i n . ~It has been found also in orchid flowers exposed t o sudden frosts4 a n d in t h e incrusting pigment of sugar cane on warming with dilute acids.5 Vanillin or a closely related mother substance, in large or small quantities, seems t o be fairly wide-spread in t h e vegetable world. Accordingly, as a possible explanation of its occurrence in field soils, it became of interest t o determine whether or not vanillin could be found in wheat as a type of field crop a n d in t h e water in which wheat had grown. V A N I L L I N I N wHEAT-Ivhen wheat seedlings were extracted with hot alcohol (recently distilled) there was obtained o n evaporating t h e alcohol extract a syrupy mass, p a r t of which was soluble in ether. T h e ether extract was treated with a half saturated solution of sodium bisulfite in t h e usual way in testing for aldehydes. T h e bisulfite solution was freed from sulfite b y means of dilute sulfuric acid a n d t h e excess of sulfur dioxide driven off by passing air through t h e liquid. After extraction with ether a n d evaporation of t h e ether solution a small a m o u n t of matter which smelled of vanillin was obtained. This gave t h e characteristic vanillin color reactions: blue with ferric chloride, green with ferrous sulfate a n d bromine water, violet with a mixture of hydrochloric a n d sulfuric acids a n d acetone water. With t h e recognition t h a t wheat contained vanillin, it became of interest t o determine how much of t h e aldehyde was present. For t h e quantitative estimation recourse was had t o t h e colorimetric method described b y Folin a n d D e n k 6 For t h e quantitative determination, zoo g. of wheat seeds were extracted with alcohol a n d t h e extract was t h e n treated in t h e customary way for extracting aldehydes. T h e final ether extract from t h e solution freed from bisulfite was evaporated a n d taken u p with warm water. This water solution was treated with equal parts of lead acetate a n d lead subacetate as long a s a precipitate formed. T h e filtrate or a n aliquot p a r t of it was treated with j cc. of t h e phosphotungsticphosphomolybdic reagent a s described by Folin and Denis a n d after 5 minutes a saturated solution of sodium carbonate was added until t h e volume of t h e liquid was 50 or I O O cc. After standing I O minutes, t h e solution was filtered a n d t h e blue color generated in t h e reactions compared in a Schreiner colorimeter with t h a t produced b y a similarly treated s t a n d a r d 1 Schreiner and Reed, Bull. 47, Bur. of Soils, U. S. Dept. Agric. (1907); Schreiner and Skinner, Bull. 77, Bur. of Soils, U. S. Dept. Agric. (1911). 2 E. C. Shorey, J . Agr. Res., 1 (19141, 357. 8 Abderhalden, Biochemische H a n d e x i k o n , 1 (19 11). 837. 4 V. Lippmann, Be*. d . chem. G6s , 46 ( 1 9 1 3 , 3431. 5 Langguth and Steuerwald. Chem. Zentr.. 1912, I, 83. 4 J . Bid. Chem., 12 (1912), 239; THISJOURNAL, 4 (1912). 670.
Vol. 6, N o .
11
solution of vanillin. The s t a n d a r d was of such a strength t h a t IO cc. diluted with t h e reagents t o I O O cc. contained I mg. of vanillin or I O parts vanillin per million. T h e stock solution of vanillin was made, a s recommended by Harder,’ by dissolving z g. of vanillin in zoo cc. of 9 j per cent alcohol a n d diluting t o I liter a n d t h e n using I O cc. of this solution t o make zoo cc. of t h e s t a n d a r d solution a s needed. Ten cc. of this standard were treated with j cc. of t h e phosphotungsticphosphomolybdic reagent a n d t h e n with sodium carbonate as outlined above. Vanillin was similarly determined ( I ) in t h e ethersoluble p a r t of a solution obtained b y heating wheat seeds 4 hours on a steam b a t h with 5 per cent sulfuric acid; ( 2 ) in the alcohol extract of wheat b r a n ; (3) in t h e alcohol extracts of t h e roots, tops, a n d seeds of wheat seedlings j t o I O days old; (4) in t h e seed water or t h e water bathing t h e seedlings for t h e first 2 4 hours; ( j ) in t h e root water or t h e water which bathed t h e roots for 4 t o I O d a y s with a change of water every two days. The water solutions were concentrated under reduced pressure a n d extracted with ether. The quantitative results are given in t h e following table: TABLEI-VANILLIN
IN
WHEAT AND I N THE WATERIN SEEDLINGS HADGROWN
WHICH
WHEAT
VAHILLIN Parts Der million MATERIAL OUANTITY 3 Wheat seeds., , . , . . , . , . . , , , , . , . , 200 and 500 g. 1-6 Wheat seeds digested with 5 yo HzSO4. 200 g. 5-14 11 3 Wheat bran.. , , . . , . . , . . . . . . . . . , , 200 g. 3(a) 13 Roots of wheat seedlings.. . . , . . , , 1500 seedlings 7-18(a) 15 Tops of wheat seedlings.. . , , , , . . , . . . 1500 seedlings 13-16(a) 7 Seeds of wheat seedlinas... . . . . . , . 1500 seedlings 6-8(s) Trace-3 Seed water., , . . , . , . ,, . ,, 4000 cc. Root water., . , . , , , , , , . . , , . , . , . . . 5000 cc. Trace to lO(a) (a) Based on weight of original seeds.
.
.
.
.. . .. . . . . . r.. .. . .. . . .
-
~
T h e effect of germination on t h e amount of vanillin in wheat may be seen more clearly in t h e following experiment where t h e vanillin in I 7 I 7 ungerminated seeds a n d in t h e same number of seeds germinated is compared: TABLE 11-VANILLIN
IN UNGERMINATED SEEDSAND IN SEEDS, TOPS, AND ROOTSOF SEEDLINGS 5 DAYSO L D P. p. rn. Seed... .. . . . . . . . . . . . . 3 . 6 Plant seed.. . . . . . . . . . . 7 . 7 Plant t o p s . , . . , , . . , . , , 1 5 . 2 (Based on weight of 1 7 1 7 seeds or 58 9.) Plant roots.. . . . . , . . . . 1 3 . 0
WHOLEP L A N T , .
..
35.9
T h e tables show t h a t vanillin occurs free in ungerminated wheat t o a slight degree, t h a t i t is increased in amount during t h e early growth of t h e plant a n d can pass from t h e plant t o t h e medium of growth either directly or more probably as a result of cell sloughing a n d disintegration. T r e a t m e n t of ground seeds with dilute sulfuric acid increases t h e a m o u n t of vanillin extractable. Accordingly, i t would seem t h a t a mother substance of vanillin occurs in wheat a n d yields vanillin on hydrolysis by acids or in t h e process of germination. Coniferin is regarded b y many as t h e mother substance of vanillin. Coniferin gives a blue color with phenol a n d hydrochloric acid especially in t h e presence of a little potassium chlorate, gives glyco-vanillin on t r e a t m e n t with potassium permanganate and chromium trioxide. yields vanillin on t r e a t m e n t with chromic 1
THISJOURNAL, 6 (1913), 619.
Kov., 1914
T H E J O U R N A L OF I N D C S T R I A L A N D E N G I N E E R I N G C H E M I S T R Y
acid mixture, yields glucose a n d a resin b y boiling with dilute sulfuric acid. F r o m t h e roots of wheat seedlings t h e r e was obtained, b y extraction m,ith h o t waterand hot alcohol and precipitation f r o m alcohol b y of ether, material which gave a greenish b l u e color
with and acid; a “lor with phenol’ acid and a little potassium chlorate; a reducing substance on boiling with dilute Sulfuric acid. T h e method of obtaining t h e material. i t s reaction with phenol, a n d t h e formation of reducing substances on boiling with acid mould indicate t h e presence of coniferin in t h e wheat roots. Coniferin, i t might be said i n passing, has been found in t h e lignified tissue of a n u m b e r of plants. Vanillin we have found also i n rotten oak wood, in pineapple pulp a n d parings a n d in t h e hot water extract of lawn grass, consisting of blue grass a n d a small a m o u n t of white clover. T h e a m o u n t of vanillin extractable from grass is considerably increased b y heating with j per cent sulfuric acid for 4 hours on a steam b a t h . Undoubtedly i t occurs t o a greater or less degree in m a n y plants either free or in a conjugated form, from which i t m a y be liberated by oxidation a n d hydrolysis. I t s presence in wood and various forms of vegetation would lead t o t h e conclusion t h a t t h e vanillin in soil has i t s origin in vegetable dkbris a n d t o a minor degree also in direct excretion or cell sloughing b y growing plants. SOIL
FERTILITY INVESTIGATIONS, BUREAUOF U. s. DEPARTXENT O F AGRICULTURE WASHINGTON
SOILS
AN EXACT AND EASY METHOD FOR PREPARING A NEUTRAL. AMMONIUM CITRATE SOLUTION By J. M. MCCANDLESS Received August 8. 1914
Since t h e writer made his official report as Referee on Phosphoric Acid. in 1908 t o t h e Association of Official Agricultural Chemists, there have appeared a n u m b e r of interesting articles on t h e subject m a t t e r of t h a t report showing how t h e a m m o n i u m citrate solution could be rendered neutral b o t h b y physical a n d chemical methods. All of this work has confirmed t h e accuracy of t h e work done, a n d t h e conclusions reached b y t h e writer in his original report. As this report was never published i n a n y chemical journal, being printed only in t h e Proceedings of t h e A. 0.A. C., a n d has therefore never been seen b y m a n y who are interested in t h e subject, t h e writer m a y be pardoned for reproducing here a p a r t of t h a t report. P A R T I A L REPRODUCTION OF REPORT OF REFEREE O N PHOSPHORIC ACID FOR
1908
“As the referee was to decide whether the various solutions
of ammonium citrate sent him were neutral or not, and as scarcely any two chemists can agree upon the exact point of neutrality, whether from lack of sensitiveness of the indicators, or color-blindness on the part of the operators, he decided to make an analysis of each sample according to the method outlined in his last report to the Association, and he guided by those results in deciding upon neutrality. METHOD OF ANALYSIS-“Twenty-five CC. O f each Solution were pipetted into a 250 cc. flask, diluted to mark, shaken, and 2 5 cc. of this solution were pipetted into a distillation flask. To the 1
Bur. of Chem , U. S. Dept. of Agr., Bull. 132, 11
921
solution in the flask, 40 cc. A7/4 caustic soda solution were added, and the contents of the flask distilled into 2 0 Cc. N / 2 acid, continuing the distillation till the Volume of the distillate measured from 6 j to 70 cc. The ammonia in the distillate was then titrated by means of N/IO alkali and cochineal. The residue in the distillation flask was washed into an Erlenmeyer, excess of standard acid added, a few drops of phenolphthalein, and the excess acid estimated by N/IO alkali. From the result, the weight of citric acid originally combined with the ammonia was calculated. Calculating from the formula of the pure salt, (NH4)3C6HjO1, that the ratio of ammonia to citric acid was as I : 3.765, a basis of comparison was established. Results obtained are given in table below: Milligrams Milliof citric grams of acid in ammonia 25 cc. of in 25 cc. diluted of diluted soluXumber solution = tion = of 2 1 / 2 cc. 2 1 / 2 cc. analvst orieinal orieinal 433.2 113.9 412.8 109.3 424.96 104.9 433.9 113.7 110.5 430.08 111.5 436.48 108.7 421.1 104.7 398.7 102.8 430.7
Ratio qf ammonia t o citric acid 1: 3.803 1: 3 , 7 7 5 1:4 . 0 5 1 1:3.816 I :3.891 133.915 1: 3.874 1: 3.808 1:4.189
Ratio in neutral salt (NHd3CsHsO; 1: 3.765 1 :3 . 7 6 5 1: 3 . 7 6 5 1:3.765 1: 3 . 7 6 5 1:3.765 1:3.765 1:3.765 1:3.765
REACTION WITH
CORALLIN N-eutral Alkaline Neutral Acid Slightly acid Arid Slightly acid Neutral Acid
“In the above table, all the solutions which showed materially more citric acid than 3.765 parts to I of ammonia, also showed a decidedly acid reaction t o corallin “It appears that some chemists prepare their ammonium citrate solution by treating the citric acid with excess of ammonia, and then leave the hot solution to neutralize itself, or finally adjust by means of red and blue litmus paper, or corallin. No. 3 in the above table was prepared in this way. The heat of the mixture not only drove off sufficient ammonia to render it neutral, hut continued and left it decidedly acid.” T h e writer has, only in t h e past few days, h a d his attention called t o t h e article of P a t t e n a n d Marti’ where in referring t o t h e solutions, analyses of which are given i n t h e above table, t h e y use t h e following language: “ T h e three solutions claimed t o be neutral a n d 168.76 g. b y McCandless contained 44.3 g . “3, C6Hg07 per liter, t h e ratio being 1:3.809. T h e results obtained b y us conclusively prove t h a t such a solution would be slightly acid, ’which fact is further substantia t e d b y t h e ratio of ammonia t o anhydrous citric acid being t o o wide.” P a t t e n a n d M a r t i h a d evidently failed t o note t h e headings of Columns j a n d 6 in t h e above table, o r t h e y would not have a t t r i b u t e d t o t h e writer t h e “CLAIM” t h a t solutions Nos. I, 4 a n d 8 were really neutral, since t h e heading of Column 6 expressly s t a t e d t h a t t h e words “neutral,” “acid” a n d “alkaline” i n t h e column below referred t o t h e reactions with corallin, a n d t h e heading of Column j showed t h a t t h e only ratio of ammonia t o citric acid which t h e referee recognized as being neutral was t h a t of I : 3.76j. T h e method given b y P a t t e n a n d h l a r t i is a n i n teresting a n d excellent method of preparing a neutral solution. T h e writer proposes the following exact a n d easy method of making a neutral solution of a m m o n i u m citrate which involves n o analyses o r distillations, b u t only a couple of titrations with solutions always r e a d y in every laboratory: 1
THISJOURNAL, 5 (1913). 568
