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Varsha Jadhav et al. / Journal of Pharmacy Research 2009, 2(4),694-697 Research Article
Available online through
www.jpronline.info
Validation of high performance liquid chromatographic method for the
determination of Tibolone in bulk and pharmaceutical dosage form
Varsha Jadhav*, Sachin Gholve and Vilasrao Kadam
*
Department of Quality Assurance, Bharati Vidyapeeth’s College of Pharmacy, Navi-Mumbai – 400614, MH, India
*For correspondence:Dr. (Mrs.) V. M. Jadhav
E-mail:drvmjadhav_bvcop@rediffmail.com
Received on:01-12-2008; Accepted on :07-02-2009
ABSTRACT
To develop and validate a sensitive, selective, precise high performance liquid chromatographic (HPLC) method of analysis for tibolone both
as bulk drug and formulations. A new sensitive and reproducible HPLC method was developed and validated for the determination of tibolone
in tablets. The separation was achieved by using a JASCO HPLC system 2000 series comprising of JASCO PU 2080 plus pump, JASCO UV
2075 plus detector, Rheodyne manual injector fitted with 100 µl sample loop. Data acquisition and treatment done using Borwin chromatog-
raphy software version 1.5. The analytical column used was thermo C8 (250 X 4.6mm; 5µm) at ambient temperature using a mobile phase
consisting of Methanol: Acetonitrile: Water (10:60:30). Flow rate was 1.0 ml with an average operating pressure of 200 kg /cm² and retention
time was found to be 6.8 ± 0.02min.The method was validated for accuracy, precision, linearity, reproducibility and robustness and statistical
comparison was performed by analysis of variance. The developed HPLC technique is precise, specific, accurate and stable. Statistical
analysis proves that the method is reproducible, selective and suitable to be applied for analysis of tibolone in commercial pharmaceutical
dosage form for routine quality control application.
INTRODUCTION
OH CH
CH3
H H
O CH3
Fig 1: Tibolone
1000000 A rea
0
0 5 10 15
Tibolone conc.m c g / m l
Fig 2: Calibration curve of Tibolone
Table 1: Data for calibration curve Table 2: Linear regression data for calibration curve
Sr. Concentration Peak Parameters HPLC method
No. of Tibolone area
(mg/ml) Range 1-11 mg/ml
1. 1 97458.87 Correlation coefficient 0.9997
2. 2 187406.75 Slope 89577x
3. 4 358744.45
4. 6 538649.89
5. 8 709397.48
6. 10 899201.45
7. 11 984456.5
Table 3: Precision of proposed HPLC method
Concentration Concentration Of Tibolone (mg/ml) found on
of Tibolone (mg/ml) Inter-day Intra-day
Mean CV Mean CV
As per the USP XXIII, system suitability tests for HPLC Linearity of the method was investigated by serially dilut-
were carried out on freshly prepared standard stock solution of ing the stock solution to give a concentration range of 1 to 11 mg/ml
Tibolone and the parameters studied and results obtained with 20 ul
injection volumes. The method was validated for accuracy, precision, and injected 20 ul with universal injector (Rheodyne). The flow rate
specificity, detection limit, quantitation limit and robustness by fol- was maintained at 1.0 ml/min. Temperature of the column was kept
lowing procedures.[3-5, 9, 10] ambient and the effluent was monitored at 205 nm. Calibration curve
was constructed by plotting concentration against peak area as shown
Accuracy: in fig 3. [3-5]