Applying green analytical chemistry (GAC) for development of

stability indicating HPLC method for determining clonazepam

and its related substances in pharmaceutical formulations and
calculating uncertainty
Abstract
Clonazepam contains one benzodiazepine ring in its chemical structure which makes it
vulnerable to degradation. In this study, green analytical chemistry approach was
applied in attempts for the development of validated stability indicating RP-HPLC
method for determining clonazepam and its related substances in pharmaceutical
formulation. Validation has been performed according to ICH guidelines. Assay was
capable of simultaneous monitoring of the intact drug in the presence of its related
substances within the same run. HPLC assay involved an ODS column and a mobile
phase composed of 2% sodium dodecyl sulfate, 0.05 M sodium acetate buffer pH 3.5
and isopropanol in ratio (25:55:20) at a flow rate of 1.5 mL/min and detection was
carried out at 254 nm. HPLC method allowed good resolution between the peaks that
corresponded to the active pharmaceutical ingredients and its degradation products with
good linearity, precision, accuracy, specificity, LOD and LOQ. The expanded uncertainty
(0.33%) of the method was also estimated from method validation data. This analytical
technique is not only ecofriendly but also faster than the conventional liquid
chromatographic system official in the USP-36.

Optoacoustic spectroscopy for real-time monitoring of strongly

light-absorbing solutions in applications to analytical chemistry
Abstract
An optoacoustic technique for solutions of strongly light-absorbing analytes at 0.1
0.01 mol l1 is proposed. The technique is based on the wide-band forward mode
detection of temporal profiles of laser-generated ultrasonic pulses (optoacoustic
signals). The leading edge of the signal repeats the distribution of the laser fluence in
the medium, which makes it possible to determine its optical absorption and investigate
its dynamics during a reaction. The range of light-absorption coefficients starts from 1 to
5 and reaches 104 to 105 cm1. The determination of iron(II) as ferroin shows the
possibility of probing 0.1 mol l1 of iron(II), which was not previously achieved for this
reaction by optical spectroscopy. To further prove the concept, kinetic measurements for
ferroin decomposition at the level of 0.1 mol l1 and at high pHs are performed. The
results are compared with spectrophotometry at lower concentrations and show good
reproducibility and accuracy of kinetic constants.