Simple Reagent Simple Reagent for Determination of Doxycycline and Tetracycline in Pharmaceuticals by UV - Visible Spectrophotometric Method

Main Article Content

Soudavone Phiphatphon
Wirat Ruengsitagoon


The objective of this work was developed for the quantification of doxycycline and tetracycline in pharmaceutical products for proposed protocol describes the use of iron ion contained in vitamin tablets as reagent. It is based on the complexation between drugs - iron ion in acidic and dilution of hydrogen peroxide. Method: This study was used commercial doxycycline and tetracycline in samples for capsules and tablets which was reacted with iron ion from vitamin tablets was concentration 5.0 ´ 10-2 mol/L in 5.0 ´ 10-3 mol/L nitric acid and 2.5 ´ 10-2  % (v/v) of hydrogen peroxide as reagent in a ratio of 2.0 : 1.0 : 0.1 (v/v), respectively. The reaction was given a yellow complex with the optimum absorption at 425 nm. Results: The calibration graphs were contained doxycycline and tetracycline standard in the range of 1.0 - 100 mg/mL and 1.0 - 200 mg/mL, respectively. Linear regression analysis of the absorbance of each drug (y) and concentration of each drug (x) expressed the equation y = 0.0064x – 0.001 (r2 = 0.9998) and y = 0.0069x - 0.0095 (r2 = 0.9997) for doxycycline and tetracycline, respectively. The limit of detection (LOD) and limit of quantitation (LOQ) each drug was 1.2, 4.0 mg/mL and 0.1, 0.4 mg/mL for doxycycline and tetracycline, respectively. The percentage relative standard deviation (RSD) of the repeatability for this proposed method was less than 2.00 %, the percentage recovery was in the range 98.62 - 100.56 % for doxycycline and 100.92 - 102.00 % for tetracycline. Conclusion: This proposed method was successfully applied in the determination of doxycycline and tetracycline in pharmaceutical products. The results obtained by the proposed method were in good agreement with method by the student t-test at the 95% confidence level.


Download data is not yet available.

Article Details

Pharmaceutical Sciences


Agwuh KN and MacGowan A. Pharmacokinetics and pharmacodynamics of the tetracyclines including glycylcyclines. J. Antimicrobial Chemotherapy. 2006; 256-265.

Al-Abachi MQ and Al-Nedawi ZA. Batch and flow injection spectrophotometric determination of doxycycline hyclate in pharmaceutical preparations. Iraqi J. Med Sci. 2015; 18: 24-32.

Anastas P, Eghbali N, Green chemistry: Principle and practice. Chem. Soc. Rev. 2010; 39:301-302.

Attia MS, Mahmoud WH, Ramsis MN< Khalil LH, Othman AM, Hashem SG and Mostafa MS. Erratum to: Spectrofluorometric Assessment of Doxycycline Hydrochloride in Pharmaceutical Tablets and Serum Sample Based on the Enhancement of the Luminescence Intensity of the Optical Sensor Sm3+ Ion. J. Fluoresc. 2011; 21:1739-1748.

Campbell NC and Hasinoff B. Iron supplements: a common cause of drug interactions. Br. J. Clin. Pharmac. 1991; 31: 251-255.

Castellanos Gil E, Van schepdael A, Roets E and Hoogmartens J. Analysis of doxycycline by capillary electrophoresis method development and validation. J Chromatogr A. 2000; 895: 43-49.

Galuszka A, Migaszewski Z and Namiesnik J. The 12 principles of green analytical chemistry and the significance mnemonic of green analytical practices. Trends Analyt Chem. 2013; 50: 78-84.

Ghaemi M and Absalan Ghodratollah. Fast removal and determination of doxycycline in water samples and honey by Fe3O4 magnetic nanoparticles. J. IRAN CHEM SOC. 2014.

Grudpan K, Hartwell SK, Wongwilai W, Grudpan Sand Lapanantnoppakhun S. Exploiting green analytical procedures for acidity and iron assays employing flow analysis with simple natural reagent extracts. Talanta. 2011; 84:1396-1400.

Hasan M, Haque M, Kobir SM and Hossain M. Quantification of doxycycline hyclate in different pharmaceutical samples by UV assay. Eur J Biol Res. 2016: 2449-8955.

ICH Expert Working Group. ICH harmonized tripartite guideline: Validation of analytical procedures: Text and methodology Q2 (R1). 2005.

Jeyabaskaran M, Prof. Rambabu C, Dr Sree Janardhanan V, Uma Maheswara Rao K, Lakshmi Maneka S and Ramesh M. HPLC method development and validation for simultaneous estimation of doxycycline and tinidazole. J. pharma creations. 2015; 2(1): 16-24.

Kogawa AC and Nunes Salgado HR. Quantification of Doxycycline Hyclate in Tablets by HPLC - UV Method. J Chromatogr S. 2000; 895: 43-49.

Mohamed HM. Green, environment-friendly, analytical tools give insights in pharmaceuticals and cosmetics analysis. Trends Anal. Chem. 2015; 66: 176-192

Newton PN, Chaulet JF, Brockman A, Chierakul W, Dondorp A, Ruangveerayuth R, Looareesuwan S, Mounier C and White NJ. Pharmacokinetics of Oral Doxycycline during Combination Treatment of Severe Falciparum Malaria. Antimicrob. Agents Chemother. 2005; 49: 1622-1625.

Palamy S and Ruengsitagoon W. A novel flow injection spectrophotometric method using plant extracts as a green reagent for the determination of doxycycline. Spectrochim Acta part A Mol Biomol. Spectrosc. 2017; 171: 200-206.

Palamy S and Ruengsitagoon W. Reverse flow injection spectrophotometric determination of ciprofloxacin in pharmaceuticals using iron from soil as a green reagent. Spectrochim Acta part A Mol Biomol. 2017; (S1386-1425917) 30752-7.

Pinyou P, Hartwell SK, Jakmunee J, Lapanantnoppakhun S and Grudpan K. Flow Injection Determination of Iron Ions with Green Tea Extracts as a Natural Chromogenic Reagent. Anal.Sci. 2010; 26:619-623.

Ramesh PJ, Basavaiah R and Rajendraprasad N. Sensitive and selective spectrophotometric assay of doxycycline hyclate in pharmaceuticals using Folin-Ciocalteu reagent. Acta Pharm. 2010; 60: 445-454.

Ramesh PJ, Basavaiah R, Divya MR, Rajendraprasad N, Vinay KB and Revanasiddappa HD. Simple UV and Visible spectrophotometric methods for the determination of doxycycline hyclate in pharmaceuticals. Anal Sci. 2011; 66: 482-489.

Rufino JL, Fernandes FCB, Ruy MS, Pezza HR and Pezza L. A simple spectrophotometric method for the determination of tetracycline and doxycycline in pharmaceutical formulations using chloramine-t. Ecol. Quim., Sao Paulo. 2010; 35(4): 139-146.

Settheeworrarit T, Hartwell SK, Lapanatnoppakhun S, Jakmunee J, Christian GD and Grudpan K. Exploiting guava leaf extract as an alternative natural reagent for flow injection determination of iron. Talanta. 2005; 68:262-267.

Sunaric SM, Mitic SS, Miletic GZ, Pavlovic AN and Naskovic-Djokic D. Determination of doxycycline in pharmaceutical based on its degradation by Cu (II)/H2O2 reagent in aqueous solution. J. Anal. Chem. 2009; 64(3): 231-237.

The British Pharmacopoeia. London: H.M. Stationary office. 2015.

The United States Pharmacopeia NF. American Pharmaceutical Association. 2016.

Vilayphone S and Ruengsitagoon W. Simple spectrophotometric method for determination of iron (III) content. Isan J. Pharm Sci. 2018;14(2): 113-121.