Author: Longenecker, Joseph Bender Title: A study of the chemistry of the 1928 Pennsylvania tobacco crop Place of Publication: Copyright Date: 1930 Master Negative Storage Number: MNS# PSt SNPaAgOSI .3 <105132>**OCLC* Form:manuscriptitem2 lnput:BMM Edit:FMD 008 ENT: 980112 TYP: s DTI: 1930 DT2: LAN: eng 035 (OCoLC)38092146 037 PSt SNPaAg031 .3 $bPreservation Office, The Pennsylvania State University, Pattee Library, University Park, PA 16802-1805 090 20 Thesis 1930m $bLonge,JB $cax+(Archival) 090 20 IVIicrofilm D344 reel 31.3 $cmc+(service copy, print master, archival master) 100 1 Longenecker, Joseph Bender. 245 12 A study of the chemistry of the 1928 Pennsylvania tobacco crop $cby Joseph B. Longenecker. 260 $c1930. 300 45 leaves $c29 cm. 502 Thesis (M.S.)--Pennsylvania State College. 504 Bibliography: leaves 38-45. 533 Microfilm $bUniversity Park, Pa. : $cPennsylvania State University $d1997. $e1 microfilm reel ; 35 mm. $f(USAIN state and local literature preservation project. Pennsylvania) $f(Pennsylvania agricultural literature on microfilm). 590 Archival master stored at National Agricultural Library, Beltsville, MD : print master stored at remote facility. 590 This item is temporarily out of the library during the filming process. If you wish to be notified when it returns, please fill out a Personal Reserve slip. The slips are available in the Rare Books Room, in the Microforms Room, and at the Circulation desk. 650 0 Tobacco $zPennsylvania. 830 0 USAIN state and local literature preservation project. $pPennsylvania. 830 0 Pennsylvania agricultural literature on microfilm. T j.i ill OX aJ A Study of the Chemistry of the 1928 rennsylvania Tobacco Crop. Subnitted to the Comnittee on Graduate Study and Advanced Decrees at The Pennsylvania State College By Joseph B. Longeneclcer In partial fulfillment of the reauire- ments for the Degree of llaster of Science. Approved: 1930 rofessor cK SoITs and Pliylochenis ^^^-^-^^^ ^^.^^^Uuuu^ Head of the Department of Agricultural and Biological Chemistry. T'^i>^v'2> TABIJC OF COIT'i^Eirj.'S I. II. III. IV. V. VI. VII. VIII. IX. Introdv.otion. RoyIqvi of Literature. 1. Fertilizer Experiment Stiidies. 2# Tjrpes of Tobacco. 3. Types of soil. 4. C\u'in{j and Fermentation Purpose and Plan of tlie Exp er in en E>rperinental Procedure. II et hods of Analysis, Presentation of Data. Discussion of Hes-alts. Aclcnov/ledsenent s . Bibliography. 4- Page 1 2 7 8 9 9 1 13 13 14 37 28 INTRODUCTION. The auality of ci^-^r tobacco depends upon the chenioal composition of the leaf at the tine it is har- vested, and upon the Icind of treatment it receives during the "nrocess of ourinf:; and fermentation. More is ]:novm about the effects of cnvironnonts on the changes which occur in the leaf during the process of curing and fer- mentation than is Icnov/n about the effects of the compo- sition of the leaf at the time it is harvested, therefore, it is of especial importance to leax^ to v/hat extent the leaf constituents govern chemical charges which occur during this important sta^^-e. The inorganic composition of a tobacco leaf is determined, to a great extent, by the type of soil on v/hich it is r-crovm. Similar Icinds of tobacco grown in different sections of the country differ greatly in com- position, size, and other characteristics; but notwith- standing the fact that the nature of the soil and climate exei't a dominant influence on composition and equality, fundamental changes in the ratios of important inorganic constituents have been obtained by fertilizer treatments. The object of this experiment is to examine the effects of ertain inorganic constituents on the fermenta- tion changes, and to study the effects of these changes on o the q^uallt^ oj? tlie leaf. Reviev; of the Literature. The inportance of fertilizer treatments is shovm by IIoss irx his oxpcrincnts with flue cured toLacco in ITorth Carolina, This particular tobacco is used x^or cit::arettes. The cured product snoiild yield a thin yellow leaf, a product that carjiot be procured unless the feeding conditions of the plant in the field are carefully controlled (1) Moss obocrved that a soil rich in nitrogen produced a leaf too dark in color, and, therefore, a soil that becomes depleted in this element during the period of maturation is desirable. In the process of caring yellow tobacco, the temperatui*e and h^omidity are re£rulated carefiilly, yet, despite this, the process cannot be carried out successfully unless the tobacco is carefully grovm for that purpose. Killebrev/ (8) in his studies on the curing of tobacco, discusses the probable importance of the basic elemeiits in the leaf. He believes the final outcome of a tobacco crop may depend on the conversion and trans- location of certain substances during; the ripening and early part of the curing stage. 1. Fertilizer Tests With Flue Cured Tobacco, Tech. iiul. No. IE, 1927. 2. Tobacco Leaf, Book. z It is thought tiiat the rate o.t v/hicii tiicGe proGGGses tal:e ]ilace depends upon the presence of excess basic clenents, a considerable amount of basic s ^bstanccs in crccess of those tied up v/ith inor^janic acid radicles. It is generally recognised that a shifting of the PH tov/ard the alkaline side speeds up the processes involved in curin^^ and feinentation. The g.uantity and aualit^ of the ash exert a great influence on the Quality of tobacco, especially in regard to smoking tobacco. The variations in the ash con- st itiients, due to clinate, soil, variety and fertilizer u.sed, run more or less parallel v;ith variations in buim, aroma and taste. But Aillebrew does not believe that one can speal: definitely of the effect of inorganic constituents on the curing processes. Nessler and Schloesing ^ ^ believe the equality of the cured product, especially with regard to bum, is determined by the basic salts in the tissue. These salts are thought to function in two v;ays: they stimulate the processes which remove injurious substances during ripening and curing. In those reactions the ratio of cellulose to the other organic substances is chi?-nged to the proper proportionality for good bum. And second, the presence I/CDifgunguersuche suToba}:,Landw.ver .Sta.29 ,309 ,1883. Ubcr die verhennlichJceit des Tabol':,Landv/. Ver.Sta. 3, 98, 1860. - -^ - of the salts in the seasoned tobac(3o catalize buxTi. (1) Haley cond^^cted a series of ei^rperiments on the catalitlc effect of certain salts on combustion and reports re- sults agree very \7ell v;ith the conclusions of earlier investdit'^ators . Durin^-j the curing of tobacco, the leaf is gradually drying out. In the meantime a series of chemical or physiolO;:ical reactions are going on. The equality of the tobacco v;ill be the resultant of the series of reactions. The changes that occiu? during the first stage are designated as the results of life processes, and the rea,otions v/hich follow the living processes are thotight to be oxidations. believes the living and post mortem changes a.re Xoew ^''^ the resiilts of enzyme activity, and the degree to v;hich these enzymes v/ill function (ass-^oming humidity and temperature are properly regulated) v/ill depend upon the conditions in v/hich the tobacco v;as grovm. The leaves of cured tobacco grov/n in dry v;eather v/ill be small, thiclc and inelastic; they v/ill btiim poorly and the smolce v/ill be strong and pungent. According to Lobv/, the production of enzymes in dry v/eather is not sufficient, or, in other v/ords,the physico-chemical malce up of the protoplasmic material is not right for the proper enzymatic reactions to take place • Tobacco grov/n in a very v/et season v/ill !• On the Relation of Potassium to iron in the Combustion Qf Gigar-Leaf-Tobacco, Science, V70, rl7 , 1929 2. Curing and Fennentation of the tJlgar Leaf, U.S.Dept. of Agri- Report Ko . 59, 1899. - 5 - not cure properly. In tliio oase, the fault is attributed to the hi;r:h acidit,^ of the leaf, a condition v/hich prevents the acciirailation of enz;^Ties. In moderately dry seasons, the protoplasnic structure has the proper physico-chemical make up to insure the carrying out of the proper reactions. A too ./Treat acidity of the juices in the leaf prevents the proper changes from tal:injjigungversuche zu Tobaks, Landv/, Ver. Station 29, 1883. • On the Relation of Potassl-um to Iron in the Conbustion of Giear-Leaf-Tobacco, Science, V 70, P17 , 1929 - 7 - of a potasGiijn base v/ill exert such a trcnenduoiis influence on combustion that it will completely cover up a retarding effect of the chlorine salts. Steece (1) made a sui^vey of the work done at tobacco e:cnerinent stations and reports that differences in the ^tjier, soluble substances, nitrogen free extracts and fibre are not traceable to fertilizer treatnents. He attributes variations in these fractions to climate, t^^ie of soil and variety of tobacco. But he does believe that the catalase activity is associated v/ith the mineral content. A parallelism is shovm to exist between the humidity of the a ir in the curing sheds, and the catalase content of the (2) ^ Steece says that it is really a response that leaf is governed largely by the fertilizer treatments. Oxidative processes bring about fundamental changes dviring curing and fermentation. The oxidative activity is said to ascend to its highest level in the period following the yellowing stage. Durin,g this time the tobacco will gain in softness and elasticity. Behrens^^^ believes that the softness and elasticity is dependent upon the production of such hygroscopic compounds as malatcs. These compounds 1. Investigations on Tobacco, with Special Reference to U.S. Report on Ag. Exp. Stat ions 1925. 2. Penn Station Bui. #196, Report 1925. 3. V/eitere Beitrage zur Kenntnis des Tobak pflanze Landw Ver.Sta.46, 1896. Quali.ty 4 '1 - 8 - are ozidation loroduots and tliis uay "be one reason v;liy it is important to prod\..3e a leaf v/ith a hifrii catalase activity. But the production of a leaf with softness and elasticity is not a guarantee of good desirable tobacco. Furthermore, tobacco, nay possess, v/hen cured, most of the desired physical properties and still be worthless as a marketable product . (1) Gamer believes the ether soluble fraction of tobacco tissues is traceable to fertilizer treatments. He also believes the substances which are responsible for the agreeable taste and pleasant odor are derived from nitro- genous compoxinds. There is a lot of evidence to be seen that lends support to this conception. Practically all tobaccos used for cigar filler are grown on soils which pro- duce a rank: growth. The leaves are deeper green and heavier than those of the tobaccos used for cigar binders and wrappers and cigarettes. In the case of the lighter tobaccos, the nitrogen content of the soil should be sufficient to support grovrth; while with the heavier tobaccos, it should be sufficient to accelerate vegetative grovrth, and thus stimulate the production of fleshy tissues. Beoaxise of the large amount of nitrogenous substances in the heavy tobacco, it is difficult to procure a fermented product that is free from siibstances v/hich tend to malce the smolce pungent. Dui'ing the 1. Tobacco Cm-ing, Farmers Bui. llo. 523. ^ C"^ ^ curing processes, "ondesirable conpoiinds are renoved, and the more there are In the leaf at the tine it is hai^este'^ , the more difficiilt the task v;ill be. Gariier suggests that the curing of cigar filler toliacGO can be facilitated by har- vesting it at the time v/hen the ripening processes have partially diminished the dense masses of chlorophyll matter. Sticcessfijl ^iroduction of tobacco depends iipon the slcill with which the natural physiological processes can be substi- tuted v/ith pathological processes. The changes which can talce place during curing are of considerable magnitude, but a failure to carry the ripening processes to the proper limit cannot be corrected by careful manipulation of the agencies involved in the curing and fermentation processes, at least our information thus far is too limited to accom- plish such corrections. There is a parallelism between the nitrogenous content of a leaf and aroma, ta,ste and nicotine content. This parallelism is the be.sis for Garner's belief that the compounds which are responsible for the aroma and taste are primary derivatives of protein constitiients in the leaf. ( 1 ) i^ Bailey and Anderson^ ^attempt to poctmate certain equalities in terms of chemical composition. They were unable to find a simple relationship between the physical and chemical standards for grading tobacco. There are a number of reasons v/hy the physical characteristics of tobacco do 1. Chemical Analyses of Tobacco from the Nitrogen Plots Report of Conn. Tobacco Station Bui. 10, 1928. - 10 ^ not check up with the chenicp.l composition. There is on iineoual distribution of substances betv;een the nidrib and web or leaf proper, and, therefore, an o.nalysls of the v/hole leaf cannot be considered as a representation of the chemical influence that is exerted on the leaf proper, it is true that an analysis of both midrib and v/eb will tell more about the distribution of the various chemical lnflu.ences, but unless the rate of movement, and the con- ditions reg^alating the movement are Icno^Tn, it v/ill be im- possible to state definitely the chemical influences that are exerted on a series of physiological reactions, and xinless this can be done, it will be a hazardous venture to explain quality in terms of composition. Killebrew and Frear both report that they have observed a movement oi inorganic substances during the ripening and curing processes. Analysis of midrib and v;eb at the time of harvesting and again when the tobacco was cured showed a one-third increase of potash in the web and a considerable decrease in calcium. Other invest i^rrators o also report a movement of salts from midrib to web and the reverse order. It is not probable that the chemist will ever be able to get perfectly concordant results in an attempt to ex- plain physiological processes on the basis of chemical analysis, 1. Tobacco Leaf, Boole 2. xj -"11- for the presence of a definite ouantity of a substance, in a reacting: systen, in one state or form nay have a totally different influence than if the sane ouantity were present in a different state or fonn. — X^ PURPOSE A] [13 i-L/uI OF TIE EXPEIiUffiNT m he object of this investigation v/as to study the effects of fertiliser treatments on the potash content of a given strain of tobacco*, and to see if there is a correlntion betv/een the absorbtion of Dotash and oalcrmnj It v/as also anticipated that the potash carrier, a sulphate, would have some effect on the sulphur content of the leaf. If it is possible to change the amounts of the principal minerals o-bsorbed by the plant, then it should be possible to determine the effects of a new distribution of inorganic substances on the cured and ferments leaves. Analyses of some of the inorganic and organic constituents of the cured and ferments leaves were made with the idea that certain q.ualities of a cigar leaf can be attributed to the com- position of the tissue. The aroma and taste of a cigar leaf arises from certain organic compounds that are volatilized during combustion of the rolled leaves in the cigar. Host of the tobacco produced in Lancaster County, Pennsylvania, gives a smoke possessing pleasant arelpatic substances, but often- times the smoke also contains pungent substances which interfere with the enjoyment of the cigar. This pungency is due to incor.plete destruction of ill smelling substances during the curing and fermentation changes, probably caused by i * HI'bsiiman Strain, Peniisylvania Broadleaf . ■• Jut, O ^' inpx^ope27 handling dtirin^^ the curing season, but it is more lilzcly cauccd "by the conditions imdor vrhlch the tobacco grew and matured, conditions as effected by rainfall, temperature and fcrtilir^or treatments. The G'^ov/ing con- ditions in the summer of 1928, in so far as weather is con- cerned, \7ere favorable, therefore, the effects of the ferti- lizer treatments should be of some importance. Materials Used. Eighteen plants carefully selected from each of ten plots, one thirtieth of an acre in area, located at Ephrata, Lancaster County, Pennsylvania, v;ere harvested at the end of the grov/ing season in 1928 and removed to the curing sheds, where they v/ere air ci;j:ed. V/hen the tobacco was cured, it was taicen from the shed and dried at room temperature. V/lien the leaves were thoroughly dried, they were separated from the midrib, finely ground and put into air tight con- tainers and ]cept for analysis. The chemical treatment of the plots from which the samples were taJcen is given in Table I. Methods of Analysis. The total carbon content of the leaves of the cured and fermented tobacco v/as determined by digesting 0,15 grams of the dried material with cone. sulphuric and Chromic acid. The chlorides and sulphuric fumes given off by the digestion mixture v/ere trapped in a silver sulphate solution and constant boiling sulphuric acid. -.14 - Table I. F_ej't j 1 i s er_ Tre^tKcnt_s_j)f_J^j3_ Exp^^^^ Plot llo. • II_amir e___ Cotton- seed Eieal Nitrate of soda Prec. Bone Carbon- ate of Potash Sulphate of Potash _ Urea *fc. * ^^ V tons/a lbs. /a lbs. /a lb s . / a lbs. /a lb 3. /a Ibs./i A-1 10 400 125 130 200 A- 2 10 400 125 130 400 A- 3 10 400 125 130 600 A- 4 10 400 125 130 310 A- 5 10 A- 6 600 187 195 300 A- 7 600 187 195 600 A- 8 600 187 195 900 A- 9 600 187 195 465 A- 10 375 460 465 57 The carbon dioxide was collected in a soda line tube. The or,''anic nitrogen v/as determined by the Kjehdal , method. The total ash was determined by sepai-ating the soluble part from the insoluble and icniting and v/eishine the fractions ceparately. The soluble portion was extracted from the ash of the cautiously ignited sample with 150 cc's of hot water, evaporated to dryness and ignited to a constant weight, at a temperature not exceeding a low red heat. " 1, The insoliible fraction v/as ignited in a platimirn cr-'j.ci'ble at a tenperature sufficiently higli to driye off the couliinecL carbon dioxide and v/eiched. The s-uin of the tv;o portions v/as calc^ilated as the total ash* The potash v/as determined by the method outlined (1) by Ames and Boltz The calcium and magnesium v/ere determined by the standard methods. (2) The sulphur v/as determined by the Bomb calori- meter method. The alkalinity of the ash v/as determined as out- lined by Haley ^ . Nasset and Olsenl*^^ The alkalinity is expressed as the number of ccs. of normal sulphuric acid needed to neutralise the ash of a 100 gram sample. The moisture v/as determined by extraction of the v/ater v/ith an evacuated sulphuric dessicator, (evacuated v/ith v/ater suction). Experiment I. The v/eb portions of the cured samples v/ere analyzed for the total ash, the soluble and insoluble fractions, and the more important ash constituents • The organic nitrogen and the total carbon v/ere determined; also the allcalinity of the soluble and insoluble parts 1* Ohio Agriculture Experiment Station Bul,#285, 2. Work done by V/. I. Zimmerman • 3. A Study of certain constituents of leaf, and their relation to the burning auality of tobacco, Plant Physiology V 3, P189, 1928. - IG - of the ash. E^rperinent II, The v;e"b portions of the fermented samples v/ere analysed for the total ash and the soluble and insoluble fractions of the total ash. The ash of tv/o of the samples were analvzed for the soluble and insoluble allcalinity. The nitrogen anc carbon v;ere determined as in the previous e:q}oriment. Table II shov/s the carbondioxide free ash in the c-oi^ed and fermented samples; including the soluble and iiisoluble fractions. Table II- A study of the ash of the cured and fermented Pennsylvania cigar filler tobacco as modified by fertiliser treatments. Soluble Ash J^ Insoluble Ash fo Total Ash fo Plot No. Cured 3.99 Fer- mented Cxired Fer- mented 10.45 Cured 17.47 Fer- mented ▲-1 5.49 13.47 15.94 A-2 4.64 6.01 11.95 10.63 16.57 16.65 A- 3 4.76 6.38 12.02 10.10 16.82 16.49 A-4 4,51 5.70 11.89 10.61 16.40 16.31 A- 5 2.79 3.77 12.85 10.28 15.63 14.55 A- 6 3.57 5.40 12.33 10.67 16.41 16.07 A-7 5.00 4.90 11.48 9.94 16.47 15.39 A-8* 5.13 12.74 17.88 A- 9 4.23 4.97 12.71 10.53 16.94 15.50 A-10 3.44 5.02 12.74 10.57 16.18 15.58 * Lost the fermented sample. I - 17 - 4 E i ' A 11 T.'iblc III shov/s the ^;:atcr inbibing pov;er of the cured and fernentcd tobacco. Thene samples were both dried at roon temperature, ground and put into air tight containers. But the samples v/ere not prepared at the sane time, and it is possible that differences in the h-onidity at the time of preparation of the samples may have some- thing to do v/ith the moisture content. It, hov/ever, is of some interest to ::nov; that v/hen the dessicated samples of the c^ared and fermented tobacco were exposed to room conditions, they both regained their original weight, and prolonged exposure would not induce greater absorbtion of moistui-e from the air of either of the samples • It is evident, as seen by the Table, that there is a slight decrease in the water embibing power brought about by fermentation. Table III A study of the moisture content of cured and fer* mented Pe:insylvania cigar filler of the 1928 crop after preparation for analysis. Plot Hxunbor _ CiirecL f" Femented A-1 5,69 4.07 A- 2 5.76 3.92 A-3 5.62 4.03 A-4 5.81 4.20 A- 5 4.92 4.30 A- 6 5.70 4.04 A-7 5.06 3.75 A- 8 A-9 A- 10 5.00 4,20 5.51 3.94 4.02 - 10 - Tiie distribution of the more inportant mineral constituents in the cured leaves as effected by ferti- lizer treatucnts is shown in Table IV. The alicalinity of the ash is listed in the sane table in order to show hov; it is effected by the distribution of the ash con- stituents. Table IV. A study of the partial inoi'^Q-^^'^i^ composition of cured cigar filler tobacco of the 1928 crop, and allcalinity of the ash, as effected by fertilizer treatments. Plot ^2_«„ CaO jo llgOf^ A-1 A- 2 A-S A- 4 A- 5 A- 6 A-7 A- 8 A- 9 7.95 7.55 7.45 7.76 7.65 8,09 6.97 7.01 7.2S A-10 7,67 Sol. Insol, AUc, Alk. Total SOgf. K20^ CCS.* Cds.* Allc. * 1.14 2.57 46.7 309.6 356,3 1,16 3.16 45,7 286.5 332.5 1.52 3.35 36.6 292.5 339.1 1.16 3.21 39,5 887.0 325.5 0,04 0,75 0,80 0,81 0,96 1,18 1,74 28,1 0,92 1.40 2.81 25,1 0,82 1,47 3,26 37.3 0.85 1.68 3.27 30.1 0.81 1.05 2,84 35,4 0,85 1,49 2.23 31.3 287,0 332,4 296,0 300.0 309.1 332.3 315.1 357.5 333.3 330.6 344.5 363.6 * CCS. of normal sixlphuric acid reouired to neutralize the alJcalinity of the ash of 100 grams of material. Table V shov;s the nitrogen and carbon content of the cured and fermented leaves. Thei'e apparently is m a small change in the IT/C ratio brought about by an in- - 19 - orease in the nitroj^cn content 4 si diirin£; fermentation i' .5 Talkie V. A study of the II/C ratio of cure^T and fernented Pennsylvania cigar filler tobacco of the 1928 crop as nodified "by fertiliser treatments . Total Carbon f» Fer- Total Nit rogen fa H/G Plot Fer- Fer- nwab e r Cured raentcd 40.74 Cured 4 . 42 nented 5.17 Cured 1:9.12 nented A-1 40.25 1:7.88 A-2 42.06 40.01 4.38 5.00 1:9.58 1:8.19 A-3 40.96 41.16 4.29 4.81 ■ 1:9.54 1:8,56 A-4 42.05 41.62 4.80 4.71 1:8.76 1:8.84 A- 5 41.87 41.17 4.47 4.95 1:9.36 1:8.32 A- 6 40.92 41.07 4.51 4.93 1:9.10 1:8.33 A- 7 40.95 41.56 4'.' 57 5.14 1:8.96 1:8.10 A-8* 42.11 4.44 1:9.48 A-9 40.07 41.36 4.51 5.18 1:8.88 1:8.00 A- 10 42.24 * 42.03 4.24 4.45 1:9.96 1:9,44 * Lost fermented sample Experiment III. Sam-oles of the fermented tobacco were used for the the filler of cigars rolled with a Connecticut' binder and Srjnatra wrapper. These cigars were scored according to their smoking cLiialitics. The factors considered were bum, fire holding capacity, color of ash, coherence of ash, aroma and taste. The writer scored the cigars carefully, but feels that tJie res"alts obtained are rather iinconvincing. The 20 - n ethod of Gcoring the snolring q.ualit?les does not tirin^'^ if I m out certain fa.ctoi'3 that aliould be enph.asi:::ecL. Table VI The scoring of clears prepared from Pennsylvania cigar filler tobacco as modified by fertiliser; treatment G and rotation of crops. Coher- Fire Color ence Holding of of Burn Capacity Ash Ash Aroma Taste Total Plot ""20 To 10" 10 25 25 100 Ntmber _ J? o_int s p p int s points points points p o int s_ _ jgoint A-1* A-2 A-3 A-4 A- 5 A-6 A-7 A-8 A- 9 A- 10 18 14 14 12 17 16 14 14 16 18 10 10 10 10 10 10 10 10 10 10 10 7 6 5 8 8 6 8 10 10 6 7 8 8 5 8 8 10 10 14 20 18 5 O 20 15 12 10 10 12 18 18 10 12 17 12 12 12 10 s 74 75 73 50 67 76 65 62 66 68 G-1 / 16 10 8 6 14 14 68 G-2 17 • 10 8 8 16 15 74 rr-3 14 • 10 8 7 14 10 63 0-4 14 10 10 6 16 15 71 G-5 14 10 8 6 15 17 70 G-6 16 10 8 15 16 72 G-7 16 10 6 8 17 15 72 G-8 17 10 6 7 18 17 75 G-9 16 10 8 8 20 20 82 G-10 16 10 10 8 15 12 71 — cjJl — ♦ The tobaccos ^^rovm in the A rjerles were {iroxin on a three year rotation -nlan. V/heat - God tobacco • / Four year rotation. Y/heat - sod - com - tobacco. Weather conditions and rainfall at Ephrata, Pennsyl- vania, for the year of 1928. ]>ariniO: the months of Api-il and Hay alternate cool and wa,rn v/eather prevailed; there v/as very little ra^infall. The nonth of Jime v/as cool and v;et. ]>urin£; the time of trans- planting^ it v/as very v/et. The physical condition of a v;et soil is very much altered by exerting a pressure eg.iial to the v;ei£;ht of a horse or tobacco planter upon it. Evidence of this has been seen in greatly reduced yields, fifty per cent reductions, yihen the tobacco has been transplanted in very v/et soil. Transplanting by hand in v/et soils v;ill not have a bad effect on the grov/th of the crop. During the month of July the v/eather v/as v/ann and frequent thunder storms supplied sufficient moisture for good grov/th. From the latter part of July until the second v/eeic in September it v/as very v/arm, and, although very little rain fell during this period, occasional thunder storms modified the drought. Cooler and dry v/eather prevailed during the latter paort of September, The nonth of October v/as also cool and dry. Table VII shov;s the rainfall per month during the year of 1928. on Tal}le VII. Ro.infall diirinc the year of 1928. January February March April May Jime July August September October Novenber Decenber 2#71 inches 3^89 2.74 4,98 1.92 9.43 3.65 4.75 3.37 0.77 1.59 0.76 TI TT tr fl II TI TT II n n TI Total 40.56 IT (Komal - 42.96) 1 -»- <-. Di3C~iission oi KesuroS i i Tlie totcl acli listed in Table II, imcLer E:cperinent T, sliov;s lov.'cr values than do most tables of total ash analysis. The reason for the low ash content is due to a modification in the method of proced'jjre for the total ash analysis. It v;as learned, during the attempt to determine the ash by the usual method, that it is very difficult to get a constant v/eight by cautiously igniting the combined v/ater solvable and insoluble fractions. I'ost of the carbonaceous material can be driven off at a low temperature. When this is done, ■:;he soluble is separated from the in- soluble ash and the latter fraction can then be ignited thoroughly without danger of driving off anj' of the volatile salts, or inclusion of carbon in the fusable ash constituents. According to the usual method of analysis, after the insoluble portion was thoroughly ignited in a platin-un cmxcible, the soluble portion v/as added to the igiiited residue, and the conbined fractions v/ere cautiously i£piioed (lo\Y red heat) and v;eished as the total or crude ash. The Pennsylv'..nia cigar filler tobacco contains much lime, and the greater part of it, v/hen ignited, is present in the oxide form. When the soluble portion of the ash is added to the ignited residue, the oxides become hydrated, and perhaps during the evaporation of the v;ater and low - 24 ^ igiiition of the anh corsiderable car*bondio:cicTe conljines v/ith the hydrated line. Because of the presence of soluble salts of potassiiun and sodium, the nixturc cannot be ignited to a sufficiently hi^^h temperature to drive off the v/ater of hydration and the combined carbondiozide. The heat applied is probably Just sufficient to drive off some of the water of hydration and carbondioxide, and this is the reason v/hy it is difficult to ^et a constant v/eisht when the combined fractions are ignited. It was thought advisa^ble to determine the two portions separately, and the total ash content obtained by this method is about four per cent lower than the ash obtained by the previous method. There can be no objections to the procedure on the grounds that some of the ash constituents were driven off by ignition at a high temperature, for the temperatui^cs applied are the same as in previous methods. The original purpose of the total ash deteriJiination of the cured and fermented samples vras to obtain a method for measiu^ing the loss in dry weight which occurs during ferm.entation. It was found that the total ash content of both samples is about the same, a condition that one would not expect to find \mdcr the circ-amstanccs. It is aT)r)arent that a loss in dry matter caiuiot be accounted for on the total ash basis, yet the heat generated and the strong odors evolved durin.^ f enientation indicate tliat losses do occar, and the fact that there is no IncreaGO in total ash is not conclnsive evidence that no losses occtir; but it does sugsest that the chan£;es v/hich acconpany f eiTiontation are more involved than v/ere ina^ined. The ash content shov/s a sliz-ht decrease in the fernentcd samples; this difference ma:^ not be of any si£jnif icance , but sonethinc has happened to the proportionality v/hich exists betv/een the soluble and insoluble ash in the cured samples during the process of fomentation. The insoluble fraction decreased v/hile the soluble fraction increased. The simplest explanation for this behavior is that part cf the insoluble ash of the cured leaf v;as converted to a soluble form dui*in^ fermenta- tion. This may be true. It seems more l0;:;lcal to account for the chan^rre on this basis than to seek an explanation on the grounds that there is a movement of ash constituents to different parts of the leaf during fermentation. But a number of investigators reported that they have observed a translocation of ash constituents, a redistribution of potash, calcium and other elements diiring the period from the time of harvesting to the end of the bull: sweating stage • In this period, Killeljrev; ^ ^states the potash content in the leaf increased one-third and the calcium decreased 1. Tobacco Leaf, Boole. - 26 - considerably. Frear^-^^ reports a novencnt of uiie saue elements in the sane direction ccrrespondlns to the direction obsei-ved by ICillebrew. A possible explanation for the reverse moTenent of the salts is suggested. The v;eb portion of the leaf consists of thin layers of tissues and the loss of moisture is considerably more rapid than from the fleshy tissues composing the midrib and the stem. During the drying processes chemical changes of a catabolic nature are tal^iing place, and some of the products formed are acidic in nature. The calcium salts of these acids are less soluble than the salts existing in the v/eb previous to the acid formation, and the acids are precipitated as the calcium salts. This condition continues until the v:eb becomes dry and the rate of acid formation approaches zero. In uhe meantime the midrib and stem are drying out much more slov;ly, and their acid fonsiation v;ill continue long after the reactions in the leaf have been slov/ed by dessication. m he concentration of the transportable calcium salts in the midrib and stem v/ill fall below that of the v/eb , and diffusion tov/ard the midrib v/ill occur. In the meantime, it does not seem reasonable that there will be a movement of salts in air dries tissues in a post mortem state. But the movement of salts need not extend more than one inch from the midrib in order 1. Pennsylvania Station Reports for 1916-1922. 27 r^'f to produce noticeable differences in the ash content of the leaf. Some attenpts have been made to study the soluble and insoluble fractions of the ash. The alizalinity studies made on the ash of c-^oi^ed and fermented leaves are of some interest. The allcalinity of the soluble portion of the ash in fermented leaves v/as one-third higher than that of the cured leaves. This agrees very v;ell v/ith the increase in the soluble ash of fermented tobacco. The soluble allca^line substances, v;hen titrated v/ith a strong acid, behave as carbonates, that is, phenol phthalein titrates one-half of the allcalinity. The allcalinity is due to a di-acid base. If "the basic substance is calcuJLated as potassium carbonate, the v;ei{^ht of the carbonate v/as eq^ual to tv/o-thirds of the viexriht of the soluble ash. One is justified in concluding, from the a.bove resi^lts, that tv/o- thirds of the soluble ash is all:aline in nature, v/hile the rest must be tied vip v;ith neutral i^adicles. The soluble portions of the ash of fermented tobacco contains practi- cally all the pota^sh that is in the tissue. If the potash foimd in the soluble ash is calculated as a carbonate, it is eq.ual to three-fourths of the total soluble ash. Since the part of the soluble ash that contributes tov/ard the alacaliiiity (on a theoretical basis) is tv;o-thirds of the total ash and the highest aaoinrit of potassi-'om carbonate that can be present in the fraction is three-fourths of the i - 28 -, total amount, it can be definitely stated that at least tv/elve per cent of the potash is present in the forn of a neutral salt. It v/as surprising to learn that the loss in the soluble ash during f ermenta.tion v/as not acconpanied by a Gorresponding decrease in the insoluble alkalinity. This Y/ould lead one to believe that there is no novenent of calci-oin from the v;eb to midrib, or a conversion of in- soluble allcaline substances to a soluble form d-oring fer- mentation. If either one occurs, the final result of the complex changes must compensate in some way for an increase in the soluble aUcalinity at the e:q?ense of the insorable alicalinity. The difficulty of explaining v/hat happened to part of the insoluble ash during fermentation might easily have been averted, if analysis of the midrib before and after fermentation had not been neglected* One ex- planation for the loss in insoluble ash which has not been considered thus far is a removal of sand and dirt from the e:cterior of the leaf during the handling of the tobacco in preparation for buHcing. The cured leaf is coated with a gummy substance. It is natural that sand and dirt would stic3: to the g-ummy substance. During the heating of the tobacco, the gummy substances are destroyed, the surface becomes less sticlcy, and particles v/hich had been adhering to it will drop off. On the other hand, there may be salts - 29 - in the rnornny secretions and these v/oiild be lost to 1 he leal' v/lien the {;uxi is disintegrated. The alicalinity of the insolu"ble ash is dxie to the calcimn in the tissues. If the calcium content of the leaf is Icnovm, it should be possible to predict v/hat per- centaf^o of the total calcium is contributing tov/ard the alicalinity by nalcing use of the allcalinity number. The amount of calcium oxide that is ecLuivalent to the cLuantity of acid used is nearly eg.ual to the total calcium content. There is, however, a small amomit of magnesium in the leaf also, and it is possible that it is responsible for a small amount of the allcalinity. But, on talcing everything into consideration, it can be definitely stated that about ninety per cent in the leaf exists in such a form v;hich, upon ignition, will be allcaline in nature • It is evident that only a small pei'centage of the calcium is in eq.uilibri-um v/ith non-combustible acid residues. Considerable worlc has been devoted to the study of the effects of certain salts in the nutrient solutions on the allcalinity of the ash of the plant tissue. Certain salts, especially chlorides, lower the allcalinity of the ash; whereas others, such as sulphate, do not have much effect on the aUcalinity. Chlorides are absorbed in pro- portion to their concentration in the nutrient solution. In soils of hlf:h concentrations of chlorides the basic - 30 ^ radiclos in the plant are tied up with chlorine, especially the Golv-blc basic siitctancGS , and the allcallnity of the ash will be lev/. In the case of sulphates, the absorp- tion of this ion is not propox»tional to the concentrations in the soil solution. The absorption of sulphates is apparently rec^alated by the sjnnthetic processes within the leaves, for plants showing similar rates of growth absorb eq.iial amounts of sulphur regardless of the q.'u.antity of available sulphates in the soil. This is true only insofar as availability of the su.lphate depends upon the amount that is applied to the soil. Reactions in the soil may tend to keep the quantity of available sulphates more or less constant, regardless of the quantity applied. In previous papers it is stated that the calcl um content of the leaf is inverselyproportional to the potash content. ITasset studied the composition of the 1925 and 1926 crops of Pennsylvania tobacco. In the dry sui:imer of 1925 the potash content was very lov/ and the calci-jxi content was rather high. In the following year, with a wet sumiiier, the potash content increased materially and the calci-^juii content decreased considerably* If the small increase in the potash absorbed is responsible for the decrease in the calcium absorbed, then it must have a profound ei'fect on the quantity of the calcium in the tissue. In the year of 1926, the potash content v/as about twenty per cent higher and the calcium content about fifteen per cent lower ol than in the previous year. The sane absorption bcliavior v/as not observed in 1928. A one hiindred per cent increase in the amo-unt of calcitun absorbed did not lower the calcrajn content at all. Table IV shows that even tiioiigh it is possible to increase the potash from 1.74 to 3.55 per cent , the hi{^h availability of line in the Hasersto\7n linestone soil overr-ales \7hat effect the potash may have on- the absorption ratio. An inspection of Tables shov/ing the inorganic content of nany kinds of tobacco grovm. on widely different soils shov/s a range in the ratio of potash to calci-oni from 1:4 to 1:0.71, There is some evidence than an inverse proportionality exists betv/een potash and calci-ujn, for in nost tobaccos the siim of these tv/o elenents in the leaf is more or less constant, being about ten per cent of the total ash. The extent to v/hich the potash in the leaf can be increased by fertilizer applications depends upon the type of soil on v/hich it is grovm. , The absorption eq-^TilibriTin nay be a function of the concentration of the siin of the txro elements in the plant sap, for there is con- siderable evidence available from absorption studies that the amount of substances tal:en up by a plant is not governed by the q.uantity of the siib stance needed for naxiniim growth. On the other hand there is a possibility that the q[uantity of potassiujn absorbed is governed by its availability. ibi application of nine mmdred pounds of potash per acre is no g^aarantee that there is plent- of it available throughotit the ,:;rov7inf? season; it nay become "fisced" in the soil. If this is true, applications of potash at definite intei-^als during the ■svoxiing season night bring about a response similar to that obtained by applications of nitrates at different intervals during growth. - 32 a - It will be no.ed. that the nitrogen content of the tobacco leaves is very high, and this may explain why the Lancaster Cotinty cigar filler produces a plangent snol:e. The nitrogen content is constant for all the sanplos. It v/ould be expected to see a parallelism betv/een yield and nitrogen content. The only constituent that varied with yield v/as potash. Apparently there is no noticeable acoixmulation of the more important con- stituents, such as nitrogen, calciuia, magnesi-om and sulphtir resulting from potash deficiency, for the dis- tribution of these substances is about the same, regardless of yield. The main purpose of this investigation was to study the effects of the inorganic composition on the changes which occur during fermentation. There are three changes in the chemical composition which accompanied fermentation. 1. There was an increase in the nitro^ren content. The nitro-carbon ratio changed from about 1:9.5 to 1:8.5. 2. There was an increase in the soluble ash and a corresponding decrease in the insoluble ash. 3. There ■./as an increase rf rf in the aUcalinity of the solu'ble ash. The magnit-ade of all the charij'^es v;as about the sane, regardless of the va