300

INDUdTRIAL A N D ENGINEERING CHEMISTRY

Vol. 17, No, 3

Analysis of Synthetic Indigo by Means of Titanous Chloride'

By F. L. English and W. S. Calcott
E. I .
DU

PONT

DB

NEMOURS & CO.. WILMINCTON, DEL.

HE analysis of indigo has been the subject of investiga- to yield the most satisfactory results. A brief survey of the tion for many years, numerous methods having ap- literature revealed the use of numerous agents other than peared in the literature. Most of these are fully de- sodium hyposulfite for the reduction of indigo or indigo scribed and criticized in the t e ~ t b o o k s , ~and consequently carmine, among which may be mentioned zinc and causticla J require no further comment here. ferrous hydroxide, stannous oxidello tin and hydrochloric The three methods which have received detailed attention acid,'l electrometrica1ly,l2 etc. I n several of these articles in this laboratory are the permanganate titration of Raw- over-reduction, beyond the stage of indigo white, was disson,4 the hyposulfite reduction described by Mullerj6and the cussed, particularly by Bayer," who states that in the reductitanous chloride method tion of indigo with tin and Preliminary report is made of a direct method for the hYdrochloric acid a green o r i g i n a l l y Suggested by titration of indigo with 0.25 N titanous chloride without compoundof stannousoxide Knecht and Hibbert.e None of these methods was with indigo white is first previous sulfonation of the sample. This method has not yet been perfected to the desired degree of accuracy, formed, which upon further found to be entirely satisfacbut the progress of the work done to date is recorded in the reduction is cmverted to a tory. The Permanganate hope that it may be of value to other investigators in the Pure yellow substance of method suffers from the disadvantage of determinsame field. undetermined composition. The development of a method depending upon sulfonaA similar statement is made ing all reducing Substances tion of the sample and titration with 0.05 N titanous concerning the action of as indigo, thus introduch3 upon errors of Up to 8 per cent, chloride is described in detail, typical results upon crude and purified synthetic indigo being given which prove the finely d i v i d e d indigo.'3 unless the indigo dhlfonic method capable of yielding results accurate within 0.2 per This suggested the Possiacid is purified. I n this event the determination is cent. bility of devising a method upon the basis of direct refairly accurate, but long drawn out and tedious, and hence hardly suitable for control duction of indigo with titanous chloride, eliminating the t i m e consuming sulfonation step common to previous reduction purposes. The hyposulfite method consistS briefly in sulfonating the methods. The probable course of such areactionis as follows: dried sample, diluting the sulfonation mass with water, and H 0 H OH titrating En aliquot in acid solution with approximately t 0.02 N sodium hyposulfite (Na28204) in an atmosphere of carbon dioxide. All steps in the procedure must be empirically standardized and rigidly adhered to, as even slight I I I 0 H OH H variations in the analytical procedure have a marked influIndigo Indigo white ence upon the results; even the rate of addition of the standard solution must be constant. The principal sources of error H OH H H involved in the process may be summarized as follows: (1) the indefiniteness of sodium hyposulfite as a reducing agent; (2) loss of sulfur dioxide due to decomposition of sodium hyposulfite by the sulfuric acid present; (3) the great H H H H effect of slight variations in manipulation, particularly the Hydroxy diindole Diindole rate of titration; (4)the instability of the standard solution; (5) extraneous oxidation resulting from air or other oxidizing Reduction beyond diindole is theoretically possible, involvagents in the reagents or carbon dioxide. I n addition, the ing saturation of the double bonds or molecular rearrangement, results cannot be interpreted upon an absolute basis, but but improbable under the proposed conditions. only in terms of an arbitrary standard indigo. The accuracy Preliminary Analysis of the method may be as high as 0.5 per cent under the best conditions, but in routine practice discrepancies of 1 per cent To test the hypothesis, analyses were made upon a dried are not uncommon. This investigation was therefore under- sample which was later shown to contain 96.7 per cent indigo. taken to develop a more reliable method less empirical in its The procedure was as follows: 0.6000 gram was dissolved in operation. 10 cc. of concentrated sulfuric acid and drowned in 50 cc. of Of all the methods previously studied, those depending water at 0" C . to obtain a fine suspension of the sample. To upon reduction to indigo white disulfonic acid had been found this was added an excess of 0.25 N titanous chloride, the I Presented before the Division of Dye Chemistry at the 68th Meeting mixture boiled in an atmosphere of carbon dioxide, cooled to

of the American Chemical Society, Ithaca, N. Y., September 8 to 13, 1924. 3 Thorpe's, Dictionary of Applied Chemistry, Vol. 111, 1918, p. 120. 8 Allen, "Commercial Organic Analysis," Vol. V, 4th ed., p. 399. J . SOC. Chem. I n d . , 18, 261 (1899). 8 B n . , 18, 2283 (1880). a "New Reduction Methods in Volumetric Analysis", 1918, p. 34; J . SOL.Dyers Colourisls, 90, 97 (1904): 91, 292 (1905). 7 The investigation of Rawson's method was conducted by H G. Tanner and W. G.Croclcett, formerly of this laboratory.

* Binz and Marx, Ibid., 99, 529 (I908J; Ann., 48,257

10

Bin2 and Rung, 2. ongew. Chcm.. 18,490,518 (1899).

(1843).

Prudhomme, Dinglns 9oZrfrch. J., 499, 546 (1878). 11 A. Bayer, Be.,1, 17 (1808). 12Zinc and Hagenbach, 2. Elekfrochem., 4, 103 (1898); I , 261 (1899); 9, 599 (1903); Goepplesroeder, J SOC. Chem. I n d . , 13, 388 (1894); Mulletus, Ber., 46, R 1007 (1893). 18 Knecht and Hibbert, 013. E L , p. 8. G

March, 1925

INDUSTRIAL A N D ENGINEERING CHEMISTRY

301

5 O - 1 0 ' C., filtered in the absence of air, the filtrate heated to boiling, and titrated hot with 0.05 Y ferric iron solution.14 During the boiling period the blue of the original indigo suspension gave place to olive green, imparted by the precipitated reduction product. The filtrate was light yellow, changing sharply to red with an excess of ferric iron. The results are given in Table I.
Table I-Preliminary Analyses of Plant Indigo b y Direct Titration 1 cc. iron = 0.2044 cc. TiClr S of Tic13 = 0.2488 INDIGO FOUND PERCENT Recovery, Boil- Calcd. O n Hy- hydroxy ing on diin- droxy diindole Assay Tic18 Fei(SO4)a time dole diindole basis Cc. Cc. Min. basis basis Per cent Exut. Gram 98.9 95.6 20 63.7 72.1 0 6000 49 90 99.1 95.8 30 63.9 71.6 0 6000 49 9 0 99.2 95.9 45 63.9 71.0 0 6000 49 90 99.5 96.2 64.1 30 46.7 0 6000 44 9 0 99.2 95.9 63.9 30 57.2 0 6500 49 92 95.9 99.2 63.9 47.2 30 0 6000 44 92

These results indicate that reduction proceeds to hydroxy diindole rather than to diindole, and were sufficiently promising to warrant further study of the method. Accordingly, the preparation of pure indigo was undertaken, the method described below being finally adopted as yielding the best product. Preparation of Pure Indigo Twenty per cent paste was dried, passed through an 80mesh sieve, and extracted in a Soxhlet apparatus with alcohol until the extraction liquor was faintly blue with no trace of red. The residue was then dried, ground, and sublimed in small portions as follows: Ten grams were placed in a 2.54-cm. (1-inch) silica tube near one end and held in place by plugs of glass wool. The far end of the tube was connected with the suction line and the near end to a carbon dioxide generator, the gas being dried by passage through twosulfuric acid towers. The tube was swept for 2 to 3 minutes, the portion containing the sample wrapped in a single layer of asbestos paper and placed in one unit of an electric combustion furnace previously heated to 800"-850' C. The projecting portion was heated slightly to prevent condensation of water. Heating was continued, with slow passage of carbon dioxide, until the red indigo vapors were completely swept from the portion of the tube originally containing the sample. The sublimate was coppery-red and of crystalline structure, some of the crystals being a centimeter or more in length. A crude yield of 65 to 70 per cent was obtained. This product was ground and extracted with alcohol as before; a large amount of a red substance being obtained in the extract. The extracted indigo was next vatted with sodium hyposulfite, 150 grams being dissolved in a solution of 100 grams of sodium hydroxide and 100 grams of sodium hyposulfite in 2500 cc. of water. The reduced solution was filtered through double-hardened filter papers and the indigo in the filtrate regenerated by treatment with air for 24 hours. The product was filtered off, washed successively with hot 1 per cent hydrochloric acid and water, and put through the sodium hyposulfite reduction process twice more. The final product was again extracted with alcohol and dried a t 140' C. It showed a theoretical nitrogen content by the Kjeldahl method and 0.12 per cent ash.
Direct Reduction Method

remaining in solution, but this hypothesis was disproved. The effect of varying the acidity of the reaction mixture was next tried. Determinations were run under the usual conditions except that varying amounts of sodium tartrate were added, the results varying inversely as the tartrate, indicating high acidity as favoring the reaction. This conclusion could not be confirmed, however, for a series of analyses in which the amount of added sulfuric acid was widely varied indicated to the contrary. I n experiments in which the reaction time was prolonged beyond about 30 minutes, a slight film of white material was observed to deposit upon the sides of the flask above the liquid level, This was assumed to be TiO(0H)t resulting from hydrolysis of titanium compounds that splashed up during boiling and came in contact with refluxing water. Hydrochloric acid was therefore substituted for sulfuric acid in the hope that this volatile acid would maintain sufficient acidity in the vapors to prevent hydrolysis. Unexpectedly, this change had a marked influence upon the reaction, the reduced compound being a bright canary yellow, with no trace of green as when sulfuric acid was used, and the reduction, being apparently much more rapid. It was further demonstrated that the addition of extra hydrochloric acid was unnecessary, that contained in the titanous chloride solution being suficient under the following revised procedure: The sample was weighed into a 300-cc. Erlenmeyer flask, 10 cc. of alcohol, 50 cc. of water, and 10 to 15 grams of small glass beads added, and the flask stoppered and shaken vigorously for a minute or so to disperse the sample thoroughly. This mixture was boiled 3 to 5 minutes, swept with carbon dioxide, 0.25 N titanous chloride added in excess, and boiled for 30 minutes. After cooling to 5O-10' C. the precipitate was filtered off in a closed system in an atmosphere of carbon dioxide and the filtrate titrated hot with 0.05 N ferric iron. To eliminate errors due to manipulation, blank determinations were made under the conditions of the indigo analysis and the extraneous oxidation was found to be 0.6 cc. of 0.05 N iron, subsequent results being corrected for this error. This process was applied to the purified indigo sample, the results of eight determinations ranging from 99.86 to 100.03 per cent. These figures apparently leave little to be desired from the standpoint of reproducibility and accuracy, but unfortunately subsequent investigation proved them to be mere coincidences resulting from the fact that the ratio of sample to excess titanous chloride was the same throughout the series. The experiments summarized in Table I1 show the effect of variation in this ratio upon the recovery of indigo.
Table 11-Effect

of Variation of Ratio of Excess Titanous Chloride to Sample Weight

1 cc. iron Iron Cc. Purified sample 49.93 12.8 49.93 31.4 49.93 60.1 49.93 119.0 Planf sample 49.93 9 9 49.93 21.0 29.5 49.93 49.93 49.5 49.93 57.8 49.93 67.0 49.93 84.0 49.93 119.7
= 0.2046 cc. Tic13 Purity Percent

1 cc. TiCla = 0.01632 gram indigo Assay TiCh Expt. Gram c c. 1 2 3 4

5 6 7 8

0.7816 0.7145 0.6145 0.4140 0.8200 0 . 7800 0.7500 0.6800 0.6500 0.6150 0,5550 0.4250

98.9 99.5 100.0 100.8 95.4 95.5 95.5 95.5 95.7 95.9 96.4 97.8

9
10

11 12

The purified material was subjected to analysis through the procedure described above, six consecutive determinations yielding results between 99.05 and 99.25 per cent. It was thought that the low results might have been due to consumption of iron by the small amount of reduced compound
' For details of the preparation, storage, standardization etc., of the 4 titanous chloride solution see THIS JOURNAL, 12, 994 (1920).

It will be noted that in the case of the purified sample the results are an almost linear function of the excess titanous chloride, whereas with the crude sample the recovery is practically constant over a fairly wide range of excess titanous chloride concentration. A marked difference in the state of subdivision of the two materials might account for this. The increased recoveries a t high titanous chloride concentrations

302

INDUSTRIAL A N D ENGINEERING CHEMISTRY

Vol. 17, No. 3

may most reasonably be attributed to over-reduction with the formation of some diindole. The limits within which constant results may be obtained upon plant material are sufficiently wide to permit the use of the method in a routine laboratory, and it is probable that the method could be empirically standardized to serve the purpose of a rapid plant control analysis. Time was not available, however, to pursue this phase of the investigation further, but it is believed that the principle upon which the method is based is sound and that a detailed study of the mechanism of the reaction would lead to the establishment of conditions under which accurate results could be obtained.
Sulfonation Method

Table IV-Effect of Acid Strength upon Recovery of Indigo 1 cc. TiCls = 0.006257gram indigo Net assay = 0.20000gram
~ULFONATION-

Strength Expt. Percent 94 94 100 100 101 101 Time Hours

--TITRATIONTiCl; Cc. 30.67 30.65 30.65 30.68 30.68 30.76

Indigo found Percent 95.97 95.91 95.91 95.97 95.97 96.25

Attention was then directed to perfecting the titration of sulfonated indigo with 0.05 N titanous chloride, upon which some preliminary work had been done previously,Is and to a comparison of this with the sodium hyposulfite method then in use in the routine laboratories. These methods consisted, briefly, in,sulfonating 1.000 gram of the dried sample with 30 cc. of 94 per cent sulfuric acid a t 55' to 60" C. for 3 hours, drowning in water, diluting to 500 cc., and titrating aliquots with either 0.05 N titanous chloride in the presence of sodium tartrate or with 0.02 N sodium hyposulfite in acid solution. The former was standardized against metanil yellow,'e the latter against pure indigo. The sulfonation method, having been designed for the sodium hyposulfite titration, was not well adapted to the 0.05 N titanous chloride procedure, particularly in the large volume of acid recommended, necessitating the use of an excessive amount of sodium tartrate. Moreover, the duration of the sulfonation seemed needlessly long. These two points were tested in a series of analyses upon a composite sample of commercial indigo in which both the time of sulfonation and the amount of acid were varied. Sulfonation was accomplished in open beakers at a temperature of 55' to 60" C. The sulfonation masses were diluted to 500 cc., 100-cc. aliquots being taken for titration. Sodium tartrate (Na&&b0~,2H~O) was added in the proportion of 25 grams per 10 cc. of sulfuric acid used in the sulfonation. The solutions were boiled 3 to 5 minutes and titrated hot under carbon dioxide to disappearance of the blue color.
T a b l e 111-Effect

Considerable variation in the strength of sulfonating acid is evidently without effect and the conclusion previously drawn in regard to the time of sulfonation is further substantiated. In the next series of experiments the temperature of sulfonation was the principal variant, the results being set forth in Table V.
Table V-Effect

Expt.

1 2 3 4
5 6 7
S

9 10 11 12

temgerature C. 60 60 60 60 60 60 75 75 75 75 75 75

of Temperature of Sulfonation u p o n Recovery of India0 1 cc. TiCls = 0.006257gram indigo Net assay = 0.20008gram Sulfonation 94% Indigo

H0 a1
Cc. 10 10 15 10 10 15 10 10 15 . 10 10 15

Time Hours 1 1 1 2 2 2 1

1 1 2 2 2

TiCla Cc. 30.63 30.70 30.70 30.65 30.69 30.61 30.68 30.64 30.65 30.47 30.44 3 " 58 0:

found Per cent 95.85 96.05 96.05 95.90 96.00 95.80 95.97 95.87 95.90 95.38 95.32 95.72

For a one-hour sulfonation a rise in temperature of 75" C. is without effect, but if the sulfonation mass is exposed to this temperature for longer periods erratic results ensue. I n experiments conducted with 10 cc. of sulfuric acid the end point in the titanous chloride titration was not quite so sharp as when 15 cc. were used. As a final check upon the method, analyses were made upon purified indigo and upon a number of commercial samples, which were analyzed also by the sodium hyposulfite method.
Table VI-Analyses

of A m o u n t of H z S O La n d T i m e of Sulfonation upon Recovery of Indigo 1 cc, TiClr 0.006257gram indigo Net assay = 0.2000 gram
I

Expt.

-SULFONATION94% Time HzSOL Hours Cc.

1 2 3 4 5 6 7 8 9

Sodium tartrate Grams 15 30 45 15 30 45 15 30 45

-TITR Water added cc.

A TION-

25 50

50

.. .. ..
..
..

50

TiClr cc. 30.63 30.70 30.70 30.55 30.65 30.69 30.61 30.65 30.68

Indigo found Per cent 95.85 96.05 96.05 95.60 95.90 96.00 95. SO 95.90 95.97

of Various Samples of Indigo by t h e 0.05 N TiCl: a n d NarSzOc Methods Indigo Indigo by TiCl; found NatSzOl Expt. cc. Per cent Per cent Purilicd sample 99.77 1 31.71 99.74 2 31.70 99.65 31.67 3 99.71 4 31.69 99.80 5 31.72 99.80 6 31.72 99.77 7 31.71 99.80 S 31.72 Average.. . 99.76 Plant sample 93.64 93.80 9 ( A ) 29.78 93.60 93.30 10 ( B ) 29.75 93.36 94.70 11 (C) 29.68 97.02 96.10 12 (D)30.83

... ...
... ... ... ... ... ...

. ...........

The time of sulfonation and the amount of acid within wide limits of variation are thus seen to exert no appreciable influence upon the recovery of indigo. Additional water was added in those titrations involving a high concentration of sodium tartrate to obtain complete solution of the sample. Sulfonations were next made with different strengths of acid, 15 cc. being used in each case, the rest of the procedure remaining unchanged except for the substitution of 30-CC. t o p s pered weighing bottles for open beakers in which to conduct the sulfonations.
This preliminary work had been conducted by Messrs. Tanner and Crockett. Calcott and English, THISJOURNAL, 1042 (1923). 16,
18

Since the purified indigo sample contained 0.12 per cent ash, the above figure represents an average recovery of not less than 99.88 per cent. Effect of Presence of Indirubin The chief organic impurity in synthetic indigo is an isomeric compound, indirubin, or indigo red:
* I

I/

-4 sample of this material, prepared by the condensation of pure indoxyl and isatin, was obtained, and its effect upon the recovery of indigo by the foregoing procedure determined.

March, 1925

I N D U S T R I A L A X D ENGINEERING C H E M I S T R Y

303

Table VII-Analysis of Indigo and Indirubin in Admixture 1 cc. TiCh = 0.006257 gram indigo N e t assay = 0.2000 gram Recoverv -...~ calcd.-as TiCla indigo Expt. cc. Per cent Purified indigo 1 31.92 99.85 2 31.87 99.70 3 31.90 99.77 Purified indigo plus 5% indirubin 4 31.90 99.77 5 31.87 99.72 6 31.84 99.65
~~

No distinction is therefore possible between indigo and indirubin by this method, the latter being also quantitatively recovered. Conclusions Upon the data presented above, the optimum conditions for the determination of indigotin by the method under investigation may be stated as follows: Weigh 1.000 gram of the dry ground sample into a 30-cc. tall-form weighing bottle,

add 15 cc. of concentrated sulfuric acid, stopper, and place in an oven a t 60"-65" C. for one hour. Cool to room temperature, drown in 400 cc. of water and dilute to 500 CC. Pipet a 100-cc. aliquot into a 300-cc. Erlenmeyer flask, add 20 grams of crystallized sodium tartrate, connect to the carbon dioxide generator and boil for 3 to 5 minutes. Sweep with carbon dioxide and titrate hot with 0.05 N titanous chloride to the last distinct color change. The end point is readily determinable within one drop, the final color change being from green to yellow. From these data it is also evident that the results are not appreciably influenced by variations in technic. The amount and strength of the sulfonating acid, the duration and temperature of sulfonation, the volume of solution titrated, and the amount of sodium tartrate may all be varied within quite wide limits without detriment. The rate of titration is also without significance. The method is capable of yielding results accurate within 0.2 per cent, on the 100 per cent basis.

Identification of Sources of Vanilla Extracts'

By C. B. Gnadinger
MCLAUGHLIN GORMLBY KINGCo;,MINNEAPOLIS, MI".

IC'DER the present standards for flavoring extracts2 the term "vanilla extract" may properly be applied to the alcoholic extract made from the beans of any variety of Vanilla planifolia Andrews. The extract trade, however, distinguishes between the several varieties, Tahiti beans being considered the least desirable and therefore commanding a much lower price than Mexican or Bourbon beans. The extract of vanillons cannot legally be sold as vanilla extract, but supplies of vanillons are usually obtainable in this country. Since the extracts of the different varieties of beans can be distinguished only by experts, who depend upon the senses of smell and taste, a chemical method for differentiating them would be advantageous. The purpose of this paper is to point out certain differences in composition between Tahiti and Bourbon beans and to describe a method for detecting the addition of extract of Tahiti beans or extract of vanillons to the better and more costly grades of vanilla extract.

Aromatic Constituents of Tahiti Beans

I n his work on Tahiti vanilla beans, Wahlbaum3 found anisyl alcohol, anisaldehyde, and anisic acid among the aromatic constituents other than vanillin. Wahlbaum did not investigate the composit,ion of Bourbon beans, and this work was accordingly undertaken by the writer. Preliminary to the examination of Bourbon beans the work of Wahlbaum on Tahiti beans was repeated, using a slightly different procedure. Five kilograms of Tahiti beans were extracted by percolation with chloroform, which was then distilled to a volume of 800 cc. The chloroform solution was distilled with steam, the chloroform separated, and the aqueous distillate extracted with ether. The chloroform and ether extracts were combined and washed with 5 per cent sodium hydroxide solution. and then with saturated sodium bisulfite solution. The chloroform-ether solution was again distilled with steam and the
1
2

Received September 24, 1924. Officeof Secretary, U. Dept. Agr., Circ. 186. S. Schimmel's Semi-Annual Report, October, 1909, p. 142.

aqueous distillate extracted with ether. This chloroformether solution yielded on evaporation 3 grams of yellowish oil having a pleasant odor and pungent taste. A portion of the oil on oxidation with dilute acid permanganate solution yielded a colorless, tasteless acid, which on recrystallizing from hot water had a melting point of 182.8' C. (corrected), compared with 184.2' C. for anisic acid. A minute quantity of the oil mixed with 2 cc. concentrated sulfuric acid gave a red color, identical with that obtained with anisyl alcohol. The refractive index a t 24" C. was 1.5411, compared with 1.5438 for anisyl alcohol prepared in the laboratory. The oil did not solidify a t 0" C. This portion of the volatile oil of Tahiti vanilla appears to consist largely of anisyl alcohol with small amounts of other compounds, probably esters. The sodium bisulfite solution washings were treated with an excess of sulfuric acid, and carbon dioxide was passed to remove the liberated sulfur dioxide. The solution was then extracted with ether and the ether evaporated spontaneously. A small amount of liquid having a pleasant odor was obtained. On treating with phloroglucin and hydrochloric acid a faint orange color was obtained, indicating the absence of piperonal and the absence of more than traces of anisaldehyde. The quantity obtained was too small for identification. The nonvolatile residue from the original steam distillation was dissolved in ether and washed nrith the 5 per cent sodium hydroxide washings mentioned above. The alkaline solution was acidified with sulfuric acid and extracted with ether. The ether solution was washed repeatedly with saturated sodium bisulfite solution to remove vanillin and evaporated to dryness. Six grams of tasteless acid were obtained. This material was purified by crystallizing three times from hot water and three times from alcohol. Its melting point was 184.1 C. (corrected), and its neutralization equivalent 151.3. The constants for anisic acid are 184.2" C. and 152, respectively. On fusing with potassium hydroxide an acid melting a t 213.2' C. was obtained. Under similar conditions anisic acid yields p-oxybenzoic acid, melting a t 213" to 214" C. The acid is therefore anisic acid.
O