• Ashok Chakravarthy V Andhra University
  • Sailaja Bbv
  • Praveen Kumar A



Objective: The objective of the present work is to develop a simple, efficient, and reproducible stability indicating reverse phase high-performance
liquid chromatographic method for simultaneous determination labetalol and its degradation products in tablet dosage forms.
Methods: The chromatographic separation of labetalol and its degradation products in tablets was carried out on Zorbax Eclipse Plus C-18
(100 × 4.6 mm, 3.5 µm) column using 0.1% trifluoroacetic acid (TFA) (v/v) in 1000 ml of water and 0.1% TFA (v/v) in 1000 ml of acetonitrile:
Methanol (1:1) by linear gradient program. Flow rate was 1.0 mL min
with a column temperature of 35°C, and detection wavelength was carried out
at 230 nm. Known impurity is well resolved from the main active drug within 14 minutes run time.
Results: The forced degradation studies were performed on labetalol tablets under acidic, basic, oxidation, thermal, humidity, and photolytic
conditions. No degradation products were observed from the forced degradation studies, and the known impurity is well resolved from the main
active drug. The method was validated in terms of specificity, linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, precision, and
robustness as per the ICH guidelines. The method was found to be linear in the range of LOQ to 120% for all the known and unknown impurities.
The LOD and LOQ values of known impurity were found between 0.3593 and 0.7187 µg mL
, and the percentage recovery values were in the range
of 95.5-105.2% at different concentration levels. Relative standard deviation for precision and intermediate precision results were found to be <5%.
The correlation coefficient found for all compounds was not <0.99. The results obtained from the validation experiments prove that the developed
method is a stability indicating method.
Conclusion: The developed method can be successfully applied for routine analysis, quality control analysis and also suitable for stability analysis of
the simultaneous determination of labetalol and its degradation products in tablet dosage forms as per the regulatory requirements.
Keywords: Labetalol, Development, Validation, Reverse phase high-performance liquid chromatography.


Download data is not yet available.



Available from:

Fahed S, Grum DF, Papadimos TJ. Labetalol infusion for refractory

hypertension causing severe hypotension and bradycardia: An issue of

patient safety. Patient Saf Surg 2008;2:13.

Louis WJ, McNeill JJ, Drummer OH. Labetalol and other vasodilator/

Beta-blocking drugs. In: Handbook of Hypertension. Amsterdam,

Netherlands: Elsevier Sciences Publishing Co.; 1988. p. 246-73.

Riva E, Mennini T, Latini R. The alpha- and beta-adrenoceptor

blocking activities of labetalol and its RR-SR (50:50) stereoisomers. Br

J Pharmacol 1991;104(4):823-8.

Robertson D, Biaggioni I. Adrenoceptor antagonist drugs. In:

Katzung BG, Masters SB, Trevor AJ, editors. Basic & Clinical

Pharmacology. 12

ed. San Francisco: McGraw-Hill Lange Medical;

p. 151-68.





standard solution

6 99.5 1.1

Percentage assay of

sample solution

Percentage 2-hydroxy


At refrigerated condition

Percentage assay of

2 99.3 0.9

154 0.148 0.01

standard solution

6 100.1 0.5

Percentage assay of

sample solution

Percentage 2-hydroxy


2 99.5 0.7

154 0.142 0.01

Koda Kimble MA, Alldredge BK. Koda Kimble and Young’s Applied

Therapeutic: The Clinical Use of Drugs. Philadelphia, PA: Lippincott

Williams & Wilkins; 2013.

MacCarthy EP, Bloomfield SS. Labetalol: A review of its pharmacology,

pharmacokinetics, clinical uses and adverse effects. Pharmacotherapy


Louis WJ, McNeil JJ, Drummer OH. Pharmacology of combined alphabeta-blockade.

I. Drugs 1984;28



Bertram KG. Basic and Clinical Pharmacology. New York: McGrawHill

Medical; 2006. p.

Richards DA, Tuckman J, Prichard BN. Assessment of alpha- and

beta-adrenoceptor blocking actions of labetalol. Br J Clin Pharmacol


Xu QA, Timothy LM. Analytical Methods for Therapeutic Drug

Monitoring and Toxicology. Hoboken, NJ: John Wiley and Sons; 2011.

Available from:


The United States Pharmacopoeia. USP-37, NF-32. Rochville, MD,

USA: United States Pharmacopeial Convention; 2014.

Jinqi Z, Lina FU, Chao L, Guogang Z. Determination of content and

related substances of labetalol hydrochloride by HPLC. Chin J Modern

Appl Pharm 2013;1:???.

Meredith PA, McSharry D, Elliott HL, Reid JL. The determination of

labetalol in plasma by highperformance liquid chromatography using

fluorescence detection. J Pharmacol Methods 1981;6(4):309-14.

Vaishali CK, Bhavika RC, Bavaskar SR, Barhate SD. Stability indicating

RP-HPLC method for determination of labetalol hydrochloride in

pharmaceutical formulation. World J Pharm Res 2015;4(4):1149-61.

Ganesan M, Nanjundan S, Rauthan KS, Eswaran K, Tripathi P.




Asian J Pharm Clin Res, Vol 9, Suppl. 2, 2016, 242-249

Chakravarthy et al.

Rapid analysis of labetalol in human plasma using liquid

chromatography- tandem mass chromatography. Int J Pharm Sci Res


European Pharmacopoeia 8.0.

ICH. Q2 (R1), Harmonized Tripartite Guideline, Validation of

Analytical Procedures: Text and Methodology. In: Proceedings of the

International Conference on Harmonization of Technical Requirements

for Registration of Pharmaceuticals for Human Use; 2005.

Snyder LR, Kirkland JJ, Glajch JL. Handbook of Practical HPLC

Method Development. 2

ed. Hoboken, NJ, USA: Wiley-Blackwell;


Basak AK, Raw AS, Al Hakim AH, Furness S, Samaan NI, Gill DS,

et al. Pharmaceutical impurities: Regulatory perspective for abbreviated

new drug applications. Adv Drug Deliv Rev 2007;59(1):64-72.

Bakshi M, Singh S. Development of validated stability-indicating assay

methods – critical review. J Pharm Biomed Anal 2002;28(6):1011-40.

United States Food and Drug Administration. Guidance for

Industry: Analytical Procedures and Methods Validation: Chemistry,

Manufacturing, and Controls Documentation. Draft Guidance.

Rockville, MD: USFDA; 2001.

Shabir GA. Validation of high-performance liquid chromatography

methods for pharmaceutical analysis. Understanding the differences

and similarities between validation requirements of the US Food and

Drug Administration, the US Pharmacopeia and the International

Conference on Harmonization. J Chromatogr A 2003;987(1-2):57-66.

Center for Drug Evaluation and Research (CDER). Reviewer Guidance:

Validation of Chromatographic Methods, CMC 3. USFDA; 1994.

Guidance for Industry, Analytical Procedures and Methods Validation

for Drugs and Biologics. Draft Guidance, U.S. Department of Health

and Human Services, Food and Drug Admin-Istration (FDA), Center

for Drug Evaluation and Research (CDER), Center for Biologics

Evaluation and Research (CBER), CMC; 2014.

Ravichandran V, Shalini S, Sundramand KM, Rajak H. Validation of

analytical methods-strategies & importance. Int J Pharm Pharm Sci


Pranshu T, Singh RP, Vikash J. Validation: A critical parameter for

quality control of pharmaceuticals. J Drug Deliv Ther 2012;2(3):34-40.

???. Validation of compendial methods. United States Pharmacopeia

, National Formulary 32. Ch. 1225.Rockville, MD, USA: The United

States Pharmacopeial Convention; 2014.

Ashok CV, Sailaja BB, Praveen KA. Development and validation of a

dissolution method for Frovatriptan tablets by reverse phase UPLC. Int

J Pharm Pharm Sci 2015;7(4):125-30.

Venkata RG, Pavani G, Supriya P, Madhavilatha N. Reverse phase

high performance liquid chromatography method development and

validation of atorvastatin in bulk drug and formulation. Asian J Pharm

Clin Res 2015;8(6):84-7.



How to Cite

V, A. C., S. Bbv, and P. Kumar A. “STABILITY INDICATING REVERSE PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR SIMULTANEOUS ESTIMATION OF LABETALOL AND ITS DEGRADATION PRODUCTS IN TABLET DOSAGE FORMS”. Asian Journal of Pharmaceutical and Clinical Research, vol. 9, no. 8, Oct. 2016, pp. 242-9, doi:10.22159/ajpcr.2016.v9s2.13687.



Original Article(s)