of 40 cc of strong ammonia water with 60 cc of alcohol, cork, shake well and often during one hour, and let stand over night. Transfer as much of the mixture as possible to a small percolator, the neck of which is provided with a purified cotton plug, receiving the percolate in a beaker or flask. Rinse the residue in the flask into the percolator with additional portions of the etherchloroform mixture, packing it down moderately with a glass rod, and continue the percolation with the same mixture until the percolate does not give an alkaloidal reaction, or at most only an opalescence when 2 cc are evaporated, and the residue is treated with N/10 hydrochloric acid, the mixture filtered and the filtrate tested with Mayer's solution. Evaporate or distil the percolate to about 10 cc at a temperature not exceeding 70° C. Transfer to a separator, and rinse the vessel with two portions of 20 cc each of ether, alternating with two portions of 10 cc each of 2 per cent sulphuric acid. Shake out, making sure that the reaction is acid, and repeat the operation three times with 15 cc of the same strength of acid, filtering the acid solutions into another separator through a 7 cm filter, and rinsing the latter with a few cubic centimeters of water. Wash the acid solution with 10 cc of ether-chloroform mixture (1 to 3 by volume), discarding the latter, and if colored repeat until no more color is acquired. Make the solution alkaline with ammonia water and shake out with four successive portions of about 20 cc each of ether-chloroform mixture (1 to 3); collect the latter in a separator, wash with 5 cc of water, transfer the ether-chloroform mixture to a tared flask or beaker, rinsing the separator with a small portion of ether-chloroform, and evaporate the solvent at a temperature not exceeding 70° C.

(a) Gravimetric. Add 3 cc of ether, evaporate and dry to constant weight at a temperature not exceeding 70° C. Report weight of alkaloidal residue. (b) Volumetric. Dissolve the residue in about 10 cc of neutral alcohol, add 25 cc of water, 3 to 15 drops of cochineal indicator solution, measure in from a burette a slight excess of N/50 sulphuric acid, mix intimately, and titrate back with N/50 potassium hydroxid. Report net amount of N/50 acid required. Aliquot method.

Into a 200 cc flask weigh 15 grams of the powdered drug, add 150 cc of etherchloroform mixture (5 to 1 by volume), cork and shake often for several minutes. Add 10 ce of ammonia water (10 per cent), shake frequently during one hour, and let stand over night. Add 15 cc of water, or sufficient to agglomerate the drug, shake, let settle a few minutes, and then decant 75 cc of the clear solution into a graduated cylinder. Transfer the solution to a separator, rinsing the cylinder with a few cubic centimeters of ether-chloroform, and shake out with 20 cc (or sufficient to render acid) of 2 per cent sulphuric acid, repeating the operation three times with 15 cc of the same strength acid, and collect the acid solutions in another separator. Wash the acid solution with 10 cc of ether-chloroform mixture (1 to 3 by volume), discarding the latter, and, if colored, repeat until no more color is acquired. Make the solution alkaline with ammonia water, and shake out with four successive portions of about 20 cc each of ether-chloroform mixture (1 to 3); collect the latter in a separator, wash with 5 ce of water, transfer the ether-chloroform mixture to a tared flask or beaker, rinsing the separator with a small portion of etherchloroform, and evaporate the solvent at a temperature not exceeding 70° C. (a) Gravimetric. Add 3 cc of ether, evaporate and dry to constant weight at a temperature not exceeding 70° C. Report weight of alkaloidal residue. (b). Volumetric. Dissolve the residue in about 10 cc of neutral alcohol, add 25 cc of water, 5 to 15 drops of cochineal indicator solution, measure in from a burette a slight excess of N/50 sulphuric acid, mix intimately, and titrate back with N/50 potassium hydroxid. Report net amount of N/50 acid required. Whenever iodeosin indicator is used, follow the directions of the United States Pharmacopoeia VIII, page 542. Solutions standardized by means of cochineal should not be used with iodeosin without running a blank for comparison, as the indicators do not give the same neutral point.

These methods and those referred to in the United States Pharmacopœia are applied to the analysis of the following drugs with the modifications noted.

ACONITE ROOT.
Method I..

U. S. Pharmacopoeia VIII, p. 28. Report the amount of N/10 acid required.

Method II.

Aliquot gravimetric. Use ether instead of ether-chloroform mixture in the final shaking out.

BELLADONNA LEAVES.

Method I.

U. S. Pharmacopoeia VIII, p. 67. Report indicator used and amount of N/10 acid required.

Aliquot volumetric.

Method II.

BELLADONNA ROOT.

Method I.

U. S. Pharmacopoeia VIII, p. 68. Report indicator used and amount of N/10 acid required.

Aliquot volumetric.

Method II.

CINCHONA BARK.

Method I.

U. S. Pharn.acopoeia VIII, p. 102. Report total and ether-soluble alkaloids. Method II.

Total extraction, gravimetric.

COCA LEAVES.

Method I.

U. S. Pharmacopoeia VIII, p. 106. Report indicator used and amount of N/10 acid required.

Method II.

Aliquot gravimetric. Use ether instead of ether-chloroform mixture through

out.

COLCHICUM CORM.
Method I.

U. S. Pharmacopœia VIII, p. 111. Report weight of alkaloidal residue.

Method II.

Exhaust 10 grams of the powdered drug by percolation with alcohol, add 25 ce of water to the solution and evaporate. Digest and stir the residue for several minutes with 25 cc of petroleum ether and transfer the liquid to a beaker, rinsing the residue with a little fresh petroleum ether. Add to the beaker 20 cc of water and evaporate the petroleum ether. Gently warm the first residue with 10 cc of water, cool, and filter the solution through a small filter into a separator, rinsing the vessel, and filter with several small portions of water and finally with the water in the beaker. Shake out the liquid with four portions of 10 cc each of chloroform and evaporate the latter in a tared dish. Add 3 cc of alcohol and evaporate, repeat the operation and dry the residue to constant weight at a temperature not exceeding 70° C. Report weight of alkaloidal residue.

COLCHICUM SEED.
Method I.

U. S. Pharmacopoeia VIII, p. 112. Report weight of alkaloidal residue.
Method II.

See Colchicum Corm. Method II.

STANDARDIZATION OF VOLUMETRIC SOLUTIONS.

In order to compare the present pharmacopoeial method for preparing volumetric solutions, a quantity of potassium bitartrate was purified according to the prescribed directions of the Pharmacopoeia. Two equal parts of this purified potassium bitartrate were weighed out, one portion ignited at a low red temper

ature in a platinum dish, dissolved in water, and carefully mixed with the second portion weighed as above. The resulting mixture was acid in reaction to phenolphthalein, indicating that some of the bitartrate was possibly lost during the ignition. Several repetitions of the above indicated that there apparently was some loss on ignition and that the method indicated in the Pharmacopoeia required further study. Recourse was then had to normal sulphuric acid prepared by the usual methods, titrated by two independent workers using several indicators, with satisfactory results, the variations ranging from the lowest for methyl orange, 0.9890, to the highest for phenolphthalein, 1.0065. This solution was used as the basis for preparing decinormal sulphuric acid.

Mr. Parker's portion of the work was practically completed when the pharmacopoeial corrections appeared, and his results are therefore based upon the original methods, except that cochineal had been substituted for hematoxylin. The standard acid employed by him, however, was found to correspond closely with that employed by the other analysts.

The results obtained are given in the following table:

Comparison of methods for the assaying of aconite, belladonna, cinchona, coca leaves, and colchicum.

Comparison of methods for the assaying of aconite, belladonna, etc.-Continued.

Residue purified by re-solution in ether, treatment with water, etc., as in the assay of colchicum corm, U. S. P., not included in average, maximum, or minimum. Residue purified as above, substituting petroleum ether for ether, not included in average, maximum, or minimum.

DISCUSSION OF RESULTS.

ACONITE LEAVES.

The leaves were assayed by the same methods as the root. The variation by the United States Pharmacopoeia method reached 51 per cent of the average value. The aliquot gravimetric method gave more uniform results, the variation amounting to 10 per cent; but titration in a few cases indicated that, as is frequently the case, the gravimetric results are high. The Squibb physiological test was obtained with a dilution of 1-80. It will be noted that the indications by this test and by assay do not correspond to the analogous results for aconite root.

ACONITE ROOT.

For aconite root, as for the leaves, the aliquot gravimetric results showed less variation than the United States Pharmacopoeia method, but the few titration results indicate that some of the former are considerably too high. The

Squibb physiological test gave a value of 1-500, or six times as great as the leaf, instead of two to three times, as might be inferred from the assays.

BELLADONNA LEAVES.

The results of Mr. Parker by the United States Pharmacopoeia method for belladonna leaves are decidedly higher than those of the other analysts, which do not vary much among themselves. The discrepancy may indicate some deterioration in the drug in the three months intervening before the lower results were obtained. All the aliquot volumetric results, which were obtained about the same time, are fairly concordant. Experiments were made to determine whether, in the extraction of the drug by ethereal solvents, the presence of a small amount of water exercises an unfavorable influence. In the pharmacoprial method a few cubic centimeters are introduced in the form of diluted ammonia water. Mr. Parker varied the method by using alcoholic ammonia of corresponding strength (about 4 per cent), also by diluting stronger ammonia water with alcohol instead of water to the same strength. The results were decidedly lower, namely, 0.298 and 0.276 per cent.

Mr. Warren employed the total extraction method also, which is similar to the pharmacopoeial method, except in the use of alcohol for diluting the ammonia. He obtained higher gravimetric results, but titration of the residues indicated that the increase was due to impurities.

Mr. Rieger tried the total extraction method, using alcohol in one test and water in another, to dilute the ammonia water. His results are low in both cases, and no conclusions can be drawn except that the residues are impure.

BELLADONNA ROOT.

In the results for belladonna root, as in those for the leaves, the best agreement is obtained by the aliquot volumetric method, and the gravimetric results are evidently vitiated by impurities. Just as with belladonna leaves, the results of experiments on the influence of small amounts of water on the extraction of the root led to no conclusion. No positive advantage was shown to accrue from the substitution of alcohol.

CINCHONA (YELLOW AND RED).

The pharmacopoeial method for cinchona gave much less variation in the results on total alkaloid than the total extraction method, but, as was the experience last year, the results on ether-soluble alkaloid are not concordant.

The change in the pharmacopoeial method, doubling the amount of solvent used in extracting the drug, involves the shaking out of 200 cc of ether-chloroform mixture with comparatively small volumes (15, 10, and 5 ce) of dilute acid, which is a tedious and difficult task. Evidently some nonalkaloidal matter passes into the acid solution in shaking out, and from that into the alkaloidal residue, which is discolored in all cases.

In determining ether-soluble alkaloid Mr. Warren filtered the ethereal solution through cotton in a small funnel.

The total extraction method fails to exhaust the marc even after obtaining 400 to 500 cc of percolate, though weak runnings give no test for alkaloids. On testing the mare, as directed last year, by boiling with normal hydrochloric acid and shaking out the acid extract, the amounts of impure alkaloid indicated in the column under "marc" were obtained.