USP 36 Chemical Tests / 〈201〉 Thin-Layer Chromatographic Identification Test 139

ness in similar containers under identical conditions, and re-

〈197〉 SPECTROPHOTOMETRIC peat the test on the residues.
IDENTIFICATION TESTS ULTRAVIOLET ABSORPTION
The reference 〈197U〉 in a monograph signifies that a test
Spectrophotometric tests contribute meaningfully toward solution and a Standard solution are examined spectropho-
the identification of many compendial chemical substances. tometrically, in 1-cm cells, over the spectral range from 200
The test procedures that follow are applicable to substances to 400 nm unless otherwise specified in the individual
that absorb IR and/or UV radiation (see Spectrophotometry monograph.
and Light-Scattering 〈851〉). Dissolve a portion of the substance under examination in
The IR absorption spectrum of a substance, compared the designated Medium to obtain a test solution having the
with that obtained concomitantly for the corresponding USP concentration specified in the monograph for Solution. Simi-
Reference Standard, provides perhaps the most conclusive larly prepare a Standard solution containing the correspond-
evidence of the identity of the substance that can be real- ing USP Reference Standard.
ized from any single test. The UV absorption spectrum, on Record and compare the spectra concomitantly obtained
the other hand, does not exhibit a high degree of specific- for the test solution and the Standard solution. Calculate
ity. Conformance with both IR absorption and UV absorp- absorptivities and/or absorbance ratios where these criteria
tion test specifications, as called for in a large proportion of are included in an individual monograph. Unless otherwise
compendial monographs, leaves little doubt, if any, regard- specified, absorbances indicated for these calculations are
ing the identity of the specimen under examination. those measured at the maximum absorbance at about the
wavelength specified in the individual monograph. Where
INFRARED ABSORPTION the absorbance is to be measured at about the specified
wavelength other than that of maximum absorbance, the
Seven methods are indicated for the preparation of previ- abbreviations (min) and (sh) are used to indicate a mini-
ously dried test specimens and Reference Standards for anal- mum and shoulder, respectively, in an absorption spectrum.
ysis. The reference 〈197K〉 in a monograph signifies that the The requirements are met if the UV absorption spectra of
substance under examination is mixed intimately with po- the test solution and the Standard solution exhibit maxima
tassium bromide. The reference 〈197M〉 in a monograph sig- and minima at the same wavelengths and absorptivities
nifies that the substance under examination is finely ground and/or absorbance ratios are within specified limits.
and dispersed in mineral oil. The reference 〈197F〉 in a mon-
ograph signifies that the substance under examination is
suspended neat between suitable (for example, sodium
chloride or potassium bromide) plates. The reference 〈197S〉
signifies that a solution of designated concentration is pre-
pared in the solvent specified in the individual monograph,
and the solution is examined in 0.1-mm cells unless a differ- 〈201〉 THIN-LAYER
ent cell path length is specified in the individual mono-
graph. The reference 〈197A〉 signifies that the substance CHROMATOGRAPHIC
under examination is intimately in contact with an internal
reflection element for attenuated total reflectance (ATR) IDENTIFICATION TEST
analysis. The reference 〈197E〉 signifies that the substance
under examination is pressed as a thin sample against a
suitable plate for IR microscopic analysis. The reference
〈197D〉 in a monograph signifies that the substance under
examination is mixed intimately with an IR-transparent ma- GENERAL PROCEDURE
terial and transferred to a sample container for diffuse reflec-
tion (DR) analysis. The ATR 〈197A〉 and the 〈197E〉 tech- The following procedure is applicable as an aid in verify-
niques can be used as alternative methods for 〈197K〉, ing the identities of many compendial drug substances as
〈197M〉, 〈197F〉, and 〈197S〉 where testing is performed such and in their respective dosage forms.
qualitatively and the Reference Standard spectra are similarly Prepare a test solution as directed in the individual mono-
obtained. graph. On a line parallel to and about 2 cm from the edge
Record the spectra of the test specimen and the corre- of a suitable thin-layer chromatographic plate, coated with a
sponding USP Reference Standard over the range from 0.25-mm layer of chromatographic silica gel mixture (see
about 2.6 µm to 15 µm (3800 cm–1 to 650 cm–1) unless oth- Chromatography 〈621〉) apply 10 µL of this solution and
erwise specified in the individual monograph. The IR absorp- 10 µL of a Standard solution prepared from the USP Refer-
tion spectrum of the preparation of the test specimen, pre- ence Standard for the drug substance being identified, in
viously dried under conditions specified for the the same solvent and at the same concentration as the test
corresponding Reference Standard unless otherwise speci- solution, unless otherwise directed in the individual mono-
fied, or unless the Reference Standard is to be used without graph. Allow the spots to dry, and develop the chromato-
drying, exhibits maxima only at the same wavelengths as gram in a solvent system consisting of a mixture of chloro-
that of a similar preparation of the corresponding USP Refer- form, methanol, and water (180:15:1), unless otherwise
ence Standard. directed in the individual monograph, until the solvent front
Differences that may be observed in the spectra so ob- has moved about three-fourths of the length of the plate.
tained sometimes are attributed to the presence of poly- Remove the plate from the developing chamber, mark the
morphs, which are not always acceptable (see Procedure solvent front, and allow the solvent to evaporate. Unless
under Spectrophotometry and Light-Scattering 〈851〉). Unless otherwise directed in the individual monograph, locate the
otherwise directed in the individual monograph, therefore, spots on the plate by examination under short-wavelength
continue as follows. If a difference appears in the IR spectra UV light. The RF value of the principal spot obtained from
of the analyte and the standard, dissolve equal portions of the test solution corresponds to that obtained from the
the test specimen and the Reference Standard in equal Standard solution.
volumes of a suitable solvent, evaporate the solution to dry-