THIN-LAYER CHROMATOGRAPHY OF AZAPHEN

M. R. Bagreeva UDC 615.214.32.074 : 543.544

Azaphen is 2 - (4 ' -methy lp ipe raz iny l - l ' ) -10-methy l -3 ,4 -d iazaphenoxaz ine dihydrochloride (I), a new ant idepress ive compound which has been synthesized in the labora tory of ant i tubercular compounds of the S. Ordzhonikdze All-Union Sc ien t i f i c -Research Institute of Pharmaceut ica l Chemis t ry [1-4].

Compound I was p repa red by the condensation of 2-chloro-10-methyl -3 ,4-d iazaphenoxazine (II) with N-methylpiperazine dihydrochloride (HI).

~v ~ f CH~

here and later r~= ~:~N~

Thin-layer chromatography on silica gel bound with gypsum has been used for the determination of the original substances in I.

Of the i0 systems examined the ~system most suitable proved to be methanol-acetone-aqueous am- monia (45 :40 :2 .5) in which Rf (1II) was 0.15, Rf (1) was 0.33, and Rf (1I) was 0.70.

When the ch romatogram was developed in UV light the diazaphenoxazine der ivat ives (I) and (II) c lear ly f luoresced; approximately a 5 pg quantity of (II) is detected. Dragendorf reagent appeared to be be t te r for the visual izat ion and gave an orange color with (I) and (t_I); the detectable minimum for (II) was 10 #g. The dihydrochloride of (Ill) did not show with Dragendorf reagent . The solution of p-benzoquinone in acetic acid and alcohol suggested for the co lo r ime t r i c determinat ion of piperazine [5] was used for the detection of (HI). In this way it was detected as a pinkish co lored spot on the chromatogram; the detectable mini- mum was 2 pg.

Thus, in o rde r to de termine the original substances in (I), 0.01 ml of a 10% aqueous solution of (I) was applied to each of two chromatographic plates (t0 x 17 cm). The dihydrochloride of (IH) was applied to the f i r s t plate as a r e fe rence m a r k e r and compound II to the second. After chromatographing in the above solvent sys tem, the f i r s t plate was t r ea t ed with the p-benzoquinone solution and the second with Drag- endorf reagent . Labora to ry and plant samples of I did not contain the original substances as impuri t ies but two other impuri t ies were detected with Rf0.25 (A) and Rf0.5 (B) (in negligible amount); only impuri ty A f luoresced in UV light and was v isual ized with Dragendorf reagent .

It was assumed that p iperazine was possibly an impuri ty in III and that the two substances (IV and V) were fo rmed f rom it under the react ion conditions for the prepara t ion of {I).

Substance (IV) was p r epa red with substance (V) as impuri ty by the condensation of (II) with piperazine .

~ - n k _ ~ ~ - ~ - ~

2F Y

S. Ordzhonikdze All-Union Sc ien t i f i c -Research Institute of Pharmaceut ica l Chemis t ry , Moscow. T r a n s - lated f rom Khimiko-Farmatsev t ichesk i i Zhurnal, Vol. 5, No. 6, pp. 59-60, Jtme, 1971. Original ar t ic le submitted May 29, 1970.

�9 1972 Consultants Bureau, a division of Plenum Publishing Corporation, 227 West 17th Street, New.York, N. Y. 10011. All rights reserved. This article cannot be reproduced for any purpose whatsoever without permission of the publisher. A copy of this article is available from the pt~blisher for $15.00.

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Substance V (Rf0.5) was detected on the chromatogram when the react ion product was applied in a 100 ~g quantity. The main react ion product was substance (IV) with Rf 0.2-0.25; f rom the p roper t i es this was analogous to impuri ty A in (I): it f luoresced in the same way in UV light and was visual ized by Drag- endorf reagent .

The content of impuri ty A in (I) was evaluated by compar ison with a r e fe rence spot of (IV) and in all of the samples did not exceed 1%.

L I T E R A T U R E C I T E D

1. French Patent No. 1576534 (1968). 2. M . D . Mashkovskii and A. I. Polezhaeva, Farmakol . i Toksikol , No. 6, 656 (1969)o 3. A . I . Polezhaeva, O. P. Vertogradova, and M. R. Bagreeva, K h im -F a rm a t s . Zh., No. 2, 59 (1970). 4. M . D . Mashkovskii, A. I. Polezhaeva, G. Ya. Avrutskii et al. , Zh. Nevropatol. i Ps ikhia t r . , 8, 1234

(1969). 5. V, Kakac and L. J. Veidelek, Handbuch der Kolorimetrie, Jena (1962).

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