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Please cite this article in press G.Sirisha et al., Analytical Method Development and Validation for the Estimation of
Trabectedin in Bulk and Parenteral Dosage Form by RP-HPLC, Indo Am. J. P. Sci, 2018; 05(03).
Preparation of the standard solution: transferred into a 100 ml amber coloured volumetric
2.0 mg of the trabectedin working standard was flask carefully without any loss of solution. The vials
accurately weighed and transferred in to 100 ml amber were rinsed thrice with 5 ml of the diluent each time.
coloured volumetric flask. 20 ml of methanol was The solution was transferred into the same volumetric
added, sonicated to dissolve and diluted to volume with flask and diluted to volume with the diluent and mix
the diluent. 5ml of above solution was transferred in to (0.02 mg/ml). Separately injected the (20µl) of diluents
a 50ml Amber coloured volumetric flask, diluted to as blank standard preparation and responses for the
volume with the diluent (0.02 mg/ml). analyte peaks were recorded.
Linearity:
The linearity of the method was established by determining the absorbance of different concentrations of Trabectedin
over a range of 50-175µg/ml respectively. The linearity data was given in table 1.
Precision:
Precision is one of the important factors which determine the reliability of an analytical method. The precision of the
developed method was tested and was found to be suitable. Both system and method precision were performed and are
given in table 3,4.
Table 3: System precision data of Trabectedin
Number of Injections RT Peak Area
1 6.661 2166730
2 6.665 2173173
3 6.662 2168564
4 6.669 2166943
5 6.672 2164318
6 6.672 2169845
Average 2168262
SD 3044.77
%RSD 0.14
Robustness:
The robustness of the proposed method was determined by analysis of aliquots from homogenous lots by differing
physical parameters like volume of injection, wavelength which may differ but the responses were still within the limits
of the assay.
Table 5 : Robustness data of Trabectedin
Where, σ = the standard deviation of the response S = the slope of theS calibration curve
S = the slope of the calibration curve The slope S may be estimated from the calibration
curve of the analyte.
The slope S may be estimated from the calibration
curve of the analyte The results of LOD and LOQ for the drug are shown in
table 8 below.
The LOD for this method was found to be 0.016 ng/ml and The LOQ for this method was found to be 0.050ng/ml.
Assay:
Assay of different parenteral formulations of Trabectedin available in the market was carried by injecting sample
corresponding to equivalent weight into HPLC system and recovery studies were carried out.
DISCUSSION: CONCLUSION:
In the present work, an attempt was made to provide a The proposed method for the assay of the Trabectedin
newer, sensitive, simple, accurate and economical RP- in the commercially available dosage formulation was
HPLC method. It was successfully applied for the simple, accurate, economical, and rapid. It can be
determination of Trabectedin in pharmaceutical dosage easily adopted for routine quality control for
forms without the interferences of other constituents in monitoring the assay in the API, in-process samples,
the formulations. Different mobile phase compositions and the finished tablet formulation.
were tried, to get good optimum results. Mobile phase
and flow rate selection was done based on peak REFERENCES:
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