The correctness of the results of the analysis of flamin for I and II were checked by

comparing the results of oscillopolarography with those of the chromatospectrophotometric

method [3]. For this purpose we took flamin of batch 126 from L'vov pharmaceutical chemicals
factory.
The results and their statistical treatment are shown in Table 2.
We analyzed for their amounts of I and II nine different batches of flamin and compared
the results obtained with the total amounts of flavonoids determined by the spectrophotometric
method of FS 42-161-72.
The results of these determinations are shown in Table 3. They
agree satisfactorily with literature data [1-3].
LITERATURE CITED
i.

2.

3.
4.

A. L. Litvinenko and V. P. Georgievskii, in: Modern Problems of Pharmaceutical Science

and Practice (Abstracts of Lectures at the Second Congress of Pharmaceutics of the
Ukrainian SSR) [in Russian], Kiev, No. 2 (1972), p. 646.
V. P. Georgievskii, Proceedings of the Second Congress of Pharmacists of the Ukrainian
SSR, Kiev (1974), p. 122.
V. P. Georgievskii, N. A, Kazarinov, A. L. Litvinenko, et al., in: Proceedings of the
Second All-Union Congress of Pharmacists [in Russian], Riga (1974), p. 163.
G. Zemplen, R. Bognar, and J. Szekely, Ber., 76, 386 (1943).

PHOTOCOLORIMETRIC

DETE~IINATION OF MANGIFERIN

S. V. Rusakova and S. I. Kocherga

UDC 615.282.074:543.432

We have previously developed a spectrophotometric method for the quantitative determination of mangiferin [i]. In the present report we describe a photocolorimetria method of
determining mangiferin, which has a number of advantages over the previous method (fewer
stages of analysis, a change of the critical solvent from tetrahydrofuran to dioxane, etCo).
The method is based on the ability of mangiferin to form complexes with ferric chloride
[2]. Such a reaction was first used for the analysis of xanthone glycosides, and mangiferin
is one of such glycosides.
Mangifer was determined from a calibration curve relating optical density of the
solution to the concentration of mangiferin.
The data were statistically analyzed.
Practically identical results were obtained on comparing the spectrocolorimetric method with the
photocolorimetric one.
EXPERIMENTAL
Determination of Mangiferin in Crystalline Powders.
Approximately i0 mg (weighed out
exactly) of mangiferin powder was dissolved in a i:i mixture of dioxane--water in a 100-ml
volumetric flask, with the volume brought up to the mark on the flask.
To a l-ml portion of
the stock solution we added 9 ml of the dioxane-water and 0.1ml of a 0.5% solution of ferric
chloride in methanol.
The optical density of the solution was determined in a photoelectrocolorimeter against a blank of i:i dioxane-water mixture, in cells of 10-mm pathlengths,
using a blue filter.
The percentage of mangiferin in the sample was calculated from a previously prepared calibration curve.
Determination of Mangiferin in Raw Materials.
The above-ground part of yellow or Alpine
French honeysuckle was crushed (particle size 0.3 mm), and 1 g was covered with methanol and
boiled for 2 h. The contents of the flask was cooled to 20~
and the extract filtered
through filter paper.
A 0.I0- to 0.15-ml sample of the solution was transferred to form a
3- to 4-cm streak on a chromatographic plate coated with a thin cellulose layer. The spot
was air-dried for 5 min, and the plate placed in a chamber for ascending chromatography, with
All-Union Scientific-Research Institute of Medicinal Plants, Moscow Province.
Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. Ii, No. 5, pp. 141-142, May, 1977.
Original article submitted July 12, 1976.

0091-150X/77/II05-0723507.50

9 1978 Plenum Publishing Corporation

723

40% acetic acid as the mobile phase.

The chromatogram was air-dried for 1 h, then examined
under ultraviolet light, and the mangiferin spots marked at Rf of 0.60 to 0.65.
(Under ultraviolet light mangiferin shows a yellowish-orange fluorescence.)
The marked cellulose
fragment was quantitatively transferred to a flask of i0- to 15-ml capacity; to it we added
i0 ml of the I:i dioxane--water mixture, and the contents were allowed to stand at room temperature for 2 h. The contents of the flask were then filtered through filter paper, and to the
filtrate we added 0.I ml of a 0.5% solution of ferric chloride in methanol.
The optical
density of the solution was determined as described above, and the percentage of mangiferin
was calculated from the previously prepared calibration curve.
Plot of the Calibration Curve.
The mangiferin standard solution was prepared by
weighing out exactly 0.05 g of mangiferin and dissolving it in 50 ml of the i:i dioxane-water
mixture.
Stock solution samples of 0.002, 0.004, 0.006, 0.008, 0.010, 0.020, 0.040, 0.060,
0.080, 0.i00, and 0.150 ml each were chromatographed, with these samples corresponding to 2,
4, 6, 8, i0, 20, 40, 60, 80, i00, and 150 ~g of mangiferin, respectively. Subsequent procedures
are outlined above.
Preparation of Plates.
Cellulose samples, 2 g each, were ground up in a porcelain mortar
with 9 ml of water, and then a uniform layer of the mixture transferred onto a 12 18 cm
glass plate.
The plate was air-dried for I0 to 15 h.
LITERATURE CITED
i.
2.
3.

B. A. Krivut, N. A. Fedyunina, S. I. Korenman, et al., Khim. Prir. No. I, 44 (1976).

S. Lewak et al., Roczn. Chem., 44, 1733 (1970).
I. M. Korenman, Photometric Analysis.
Methods'of Determination of Organic Compounds
Russian], Moscow (1975)~ pp. 264-266.

~THODS

[in

OF ACCELERATED DETECTION OF VIABILE MICROORGANISMS

IN RADIOPHARMACEUTICAL

PREPARATIONS AND IN THEIR MANUFACTURE

L. M. Kotlyarov, E. P. Pavlov,
E. G. Tushov, and V. V. Sedov

UDC 615.849.2.011.17:576.8

In the manufacture of radiopharmaceutical preparations (RPP), in a number of cases it is

required to carry out an accelerated detection of viable microorganisms, especially in the
output of RPP which contain isotopes having a short half-life.
This concerns primarily
checking for the observance of the sterilization regime of certain RPP in an autoclave by use
of a biological test (Baacilus subtilis 720), and determining the extent of "seeding" of the
manufacturing sites, working chambers, solutions of semifinished products, and the RPP with
microflora of the air before their sterilization.
Moreover, the presently known methods of
accelerated detection of viable microorganisms were not designed for use under the conditions
of radiochemical manufacturing.
With consideration of the specifics of this type of manufacturing, we selected methods of detecting microorganisms from the growth of microcolonies
on a thin layer of growth medium [i, 2] and methods of detecting microorganisms from their
biochemical activity [3-5].
The use of these methods with the objective of detecting viable
microorganisms in RPP and in their manufacture encountered definite hindrances, associated
with the fact that the times for performing various tests as a function of the content of
microorganisms in the medium were not known.
In our investigations we prepared suspensions Of various microorganisms (Bac. subtilis,
strain 720; Bac. anticoides, strain 250; Escherichia coli, strain 675; Staphlococcus aureus,
strain 209P) in solutions of RPP and in their individual ingredients.
In experiments on
detection of microcolonies, we applied the sample being tested onto a thin layer of agarized
growth medium, uniformly distributing the materials over the surface of the medium.
On the
Institute of Biophysics, Ministry of Public Health of the USSR, Moscow. Translated
from Khimiko-Farmatsevticheskii Zhurnal, Vol. ii, No. 5, pp. 142-145, May, 1977. Original
article submitted July 5, 1976.

724

0091-150X/77/II05-0724507.50

9 1978 Plenum Publishing Corporation