Simultaneous Estimation of Tinidazole Containing Combination Formulation by RP-HPLC Technique. 

M. Pharm Dissertation Protocol Submitted to

Rajiv Gandhi University of Health Sciences, KarnatakaBangalore – 560 041

ByMr. Ajay Kumar.T. B.Pharm

Under the Guidance ofDr. S. Meena,

Head, Department of Pharmaceutical Analysis.

Acharya & B.M. Reddy College of Pharmacy,

Acharya Dr.Sarvepalli Radhakrishna Road,

Soladevanahalli, Chikkabanavara (post),

Bangalore – 560090.

2012 – 2014.

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA

ANNEXURE – II

PROFORMA FOR REGISTRATION OF SUBJECTS

FOR DISSERTATION

1.0 NAME OF THE

CANDIDATE AND

ADDRESS

Ajay Kumar.T. ,#1, 1st Cross, 3rd Block, Nagarbhavi 1st Stage,

Bangalore: 560072.

2.0 NAME OF THE

INSTITUTION

ACHARYA & B.M. REDDY COLLEGE

OF PHARMACY,

Acharya Dr. Sarvepalli Radhakrishnan Road,

Soldevanahalli, Chikkabanavara (post),

Bangalore-560 090.

3.0 COURSE OF STUDY

AND SUBJECT

M. PHARMACY

(Pharmaceutical Analysis)

4.0 DATE OF

ADMISSION

30th May, 2012.

5.0 TITLE OF THE TOPIC

Simultaneous Estimation of Tinidazole Containing Combination

Formulation by RP-HPLC Technique. 

6.0

6.1:-

BRIEF RESUME OF THE INTENDED WORK:

NEED FOR PRESENT STUDY:

Tinidazole is official in IP1, BP2, and USP.3 Few combinations containing tinidazole with

Ofloxacin or clotrimazole or fluconazole are considered for the present study. The

individual drugs are official and their combinations are being introduced in the market

for the treatment of mixed aerobic/anaerobic polymicrobial infections, fungal infections

and to eradicate H.pylori infections.4,5

Studies on estimation of tinidazole have been reported by several methods.

Limited methods are available for the estimation of tinidazole in combination

formulations. Reports on simultaneous estimation of the drug combinations with

tinidazole and estimation of them by a liquid chromatographic method, validated as per

ICH guidelines, are very few and there is a need for simpler and sensitive methods to be

developed for the estimations in combinations.

In this study, attempts will be made to develop a HPLC method and validate as per ICH

guidelines, the analytical methods for tinidazole as one of the drug with its combination

such as clotrimazole or fluconazole or ofloxacin in marketed formulation.

6.2:-DRUG PROFILE:

TINIDAZOLE:

It is an anti-parasitic drug used against protozoan infections. It is widely known throughout

Europe and the developing world as treatment for variety of amoebic and parasitic infection.6

IUPAC Name 1-(2-ethyl sulfonylethyl)-2-methyl-5-nitro-imidazole.6

Solubility Soluble in acetone, methylene chloride.6

Sparingly soluble in methanol. Practically insoluble in water.

pka 4.7 (negative logarithm of the acid dissociation constant)6

Log P 0.7 (logarithm of the ratio of the concentrations unionized solute in the

solvents)6

TINIDAZOLE AND ITS COMBINATIONS:

Tinidazole with clotrimazole. (Tinidazole 500 mg and clotrimazole 200 mg).

Tinidazole with fluconazole. (Tinidazole 1 gm and fluconazole 150 mg).

Tinidazole with ofloxacin. (Tinidazole 600 mg and ofloxacin 200 mg).

6.3:- OBJECTIVES OF THE STUDY:

The aim of our study is:

To develop a liquid chromatographic (LC) method for the simultaneous estimation of

tinidazole and its combinations like clotrimazole or fluconazole or ofloxacin.

To validate the LC method developed, as per ICH guidelines, for parameters such as

accuracy, precision, specificity, robustness, linearity etc.

To study the suitability of the LC method developed for analyzing a marketed dosage

form containing tinidazole along with clotrimazole or fluconazole or ofloxacin.

7.0

7.1:-

7.2:-

MATERIALS AND METHODS:

SOURCE OF DATA:

The data will be obtained from:

JOURNALS :

Indian Journal of Pharmaceutical Sciences.

International Journal of Pharmaceutical Sciences.

International Journal of Pharm Tech Research.

IJPI’S Journal of Analytical Chemistry.

Thai Journal of Pharmaceutical Science.

Journal of Chromatographic Science.

Chronicles of Young Scientists.

www.sphinxsai.com

www.google.com

www.wikepedia.org

www.ethesis.helsniki.fi

www.drugs.com

www.omicsonline.org

www.ijppsjournal.com

www.medindia.net

www.medicinenet.com

www.drugsupdate.com

METHOD OF COLLECTION OF DATA:

Methodology of the study:

A HPLC method for simultaneous estimation of tinidazole and its combinations will be

developed.

A suitable chromatographic condition has to be optimized such as: Stationary phase,

mobile phase, flow rate, detection wavelength.

Bulk drug samples of tinidazole and its combinations shall be procured as gift sample

from industries involved in manufacture of them or from suppliers.

The analytical method development and validation study will be carried out in the

Department of Pharmaceutical Analysis, Acharya & B.M Reddy College of Pharmacy,

Bangalore-90.

.

LC method will be validated, as per ICH Guidelines,15 for following parameters:

1) PRECISION:

The response to repeated injections of the same standard solution should have RSD ≤

1.0%.

Intra assay: Injection of sample from each solution should have RSD ≤ 2.0%.

Intermediate precision: Assay of the same sample have to be carried out several times

within two to three days using same experimental conditions but a different analyst and

instrument, RSD ≤ 2.0%.

2) ACCURACY:

Direct comparison to reference standard.

Recovery studies are to be performed by spiking the blank matrix of the sample at

different level (50, 75,100,125,150%) of the known level in the sample. Average

recovery of the analyte should be in the range of 98.0-102.0%.

3) ROBUSTNESS:

The method must be robust enough to withstand such slight changes and allow routine

analysis of the sample.

Conditions such as temperature ±5ºC, buffer pH±0.5, and ionic strength of buffers and

level of additives to mobile phase have to be slightly altered.

4) LINEARITY:

Linearity is the degree to which the response measured by the method is directly

proportional to the concentration of the analyte.

By covering the range 0-150% of the expected level of the analyte.

The method must exhibit linearity in the described range.

5) RANGE :

Linearity when applied to samples containing analyte at the extremes of the range as well

as within the range. Analytical procedure should be confirmed to establish an acceptable

degree of linearity, accuracy and precision when applied to samples containing amounts

of analyte within or at the extremes of specified range.

6) SPECIFICITY:

Specificity should be conducted during the validation of identification tests, the

determination of impurities, and the assay. To determine this, the sample have to be

injected and monitored for the identification of decomposition/degradation products,

intermediates, solvents, chemicals used in analytes should separate from main analyte

with Rs not less than 3.0.

Injecting separately different components of the matrix of the sample (tablets, syrup,

suspension) should clearly separate from analyte with Rs ≥2.0.

Tinidazole and its combinations will be analyzed by the LC method developed.

7.3:- DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO BE

CONDUCTED ON THE PATIENTS OR OTHER HUMAN/ANIMALS? IF SO, PLEASE

DESCRIBE BRIEFLY.

- No -

7.4:- HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN

CASE OF 7.3?

- No -

8.0 REFERENCES:

1. Indian Pharmacopoeia, III. Ghaziabad: The Indian Pharmacopoeia Commission; 6th

ed. 2010 Sept. 2226-27.

2. British Pharamcopoeia. London: British Pharmacopoeia Commission; 2009 Jan,

2037-38.

3. U.S. Pharmacopoeia and National Formulary. Rockville, MD: The United States

Pharmacopoeia, 34th ed. 2011 May, 4446.

4. K.D.Tripathi, Essentials of Medical Pharmacology, 6th ed, New Delhi: Jaypee

Brothers Medical Publishers (P) Ltd; 797-807.

5. Horatio B. Fung, Thien-Ly Doan. A nitroimidazole anti-protozoal agent. Clinical

Therapeutics. 2005; Dec; 27(12): 1859-81.

6. Beale M. John, Block H, John Wilson and Gisvold’s Textbook of Organic Medicinal

and Pharmaceutical Chemistry, 12th ed, New Delhi: Lippincott Williams & Wilkins;

2011, 981.

7. Parul Parmar and Ankitha Mehta. Determination & validation of HPTLC method for

the estimation of clotrimazole in bulk drug tablet formulation. Ind J Pharm Sci: 2009;

71 (4): 451-54.

8. Bharath S A, Arshad M D, Venugopal Darak and Kalyan Chakravarthy K.

Development and validation of simultaneous spectrophotometric estimation of

clotrimazole & tinidazole in tablet dosage form. IJPI’S J Anal Chem: 2011; Vol 1(2):

13-17.

9. Meshram D B, Bagade S B and Tajne M R. simple HPLC method for simultaneous

estimation of fluconazole & tinidazole in combined dose tablet. J Chromatogr Sci:

2009; 47: 885-8.

10. Chiranjeevi Bodepudi, Swati Bantu, Kalyan Obula Reddy M, Shanmugasundaram P,

Vijey Anandhi M. Novel RP-HPLC method development & validation of fluconazole

& tinidazole in a combined tablet dosage form. Int J Chem Tech Res: 2011; 3(3):

1309-17.

11. Saurab Pandey, Preeti Pandey, Shaifali Dubey, Udisha Chaturvedi, Awani k Rai.

Simultaneous estimation of tinidazole & fluconazole in combined tablet dosage form.

Thai J. Pharm. Sci: 2012; 55-62.

12. Kaaja Pasha, Simultaneous determination of ofloxacin and tinidazole by instrumental

methods [M.Pharmacy], Bangalore, RGUHS, Feb 2005.

13. Dharuman J, Vasudevan M, Somasekaran K N, Dhandupani B, Prashant D Ghode &

M. Thiagarajan. RP-HPLC method development & validation for the simultaneous

estimation of ofloxacin and tinidazole in tablets. Int J Pharm Tech Res: 2009; 1(2):

121-124.

14. Kareti Srinivas Rao, Arijit Banerjee, Nargesh Kumar Keshar. Spectrophotometric

methods for the simultaneous estimation of ofloxacin & tinidazole in bulk and

pharmaceutical dosage form. Chronicles of Young Scientists. 2011; 2(2): 98-101.

15. ICH topic Q 2 B Validation of analytical procedures – methodology [internet]. 1996

[cited 2011 Mar 5]. Available from: http://Www.Uam.Es/Personal_Pas/Txrf/Mu5.pdf

16. Sethi P D. High performance liquid chromatography quantitative analysis of

pharmaceutical formulations.1st ed. New Delhi: CBS Publishers and Distributors

2001: 118-20.

17. Kasture A V, Wadodkar S G, Mahadik K R, More H N. Pharmaceutical Analysis.

19th ed. Pune, Nirali Prakashan, 2010: 48-57.

9.0 SIGNATURE OF THE CANDIDATE

10.0 REMARKS OF THE GUIDE Recommended for Research

11.0 NAME AND DESIGNATION OF:

11.1 GUIDE Dr. S. Meena , Head, Department Of Pharmaceutical Analysis,Acharya & B.M. Reddy College of Pharmacy, Soldevanahalli, Bangalore-90

11.2 SIGNATURE

11.3 CO-GUIDE NIL

11.4 SIGNATURE NIL

11.5 HEAD OF THE DEPARTMENT

Dr. S. Meena, Head, Department Of Pharmaceutical Analysis,Acharya & B.M. Reddy College of Pharmacy, Soldevanahalli, Bangalore-90

11.6 SIGNATURE

12.0 12.1 REMARKS OF THE PRINCIPAL

12.2 NAME OF THE PRINCIPAL Dr. Divakar GoliPrincipal,Acharya & B.M. Reddy College of Pharmacy, Soldevanahalli,Bangalore-90.

12.3 SIGNATURE