Author(s):
Bhaskar Mangalapu, B. Srinivas, Sirigiri Ramathulasi, Ramesha Andagar Ramakrishna, Venkata Narayana Palakollu
Email(s):
dr.pvnarayana@gmail.com , venkatanarayana.palakollu@reva.edu.in , dr.ramesha@flowchempharma.com
DOI:
10.52711/0974-360X.2025.00537 
Address:
Bhaskar Mangalapu1,2, B. Srinivas3, Sirigiri Ramathulasi4, Ramesha Andagar Ramakrishna2*, Venkata Narayana Palakollu1*
1Department of Chemistry, School of Applied Sciences REVA University, Yelahanka, Bengaluru, 560064 India. 2Flowchem Pharma Pvt Ltd., Hindupur, Andhra Pradesh., 515211, India.
3Trroy Life Sciences Pvt Ltd., Yelahanka, Bengaluru, 560064, India.
4Annamacharya Institute of Technology and Sciences, Tirupathi, Andhra Pradesh, 517520, India.
*Corresponding Author
Published In:
Volume - 18,
Issue - 8,
Year - 2025
ABSTRACT:
Anxiolytic and antidepressant uses for melitracen hydrochloride are the main ones. It helps treat anxiety and depression symptoms by adjusting the activity of certain neurotransmitters, mainly norepinephrine and serotonin. To increase the treatment's therapeutic efficacy, it is frequently used in combination with other drugs. A simple, specific, and accurate stability indication of the RP HPLC method was developed to quantify melitracen hydrochloride (MH) and its associated process impurities, aiming to provide a precise and reliable analytical tool for this bulk drug. This method adheres to the International Conference on Harmonization Guidelines (ICH). The X-Terra RP 18 column from Waters Corporation (15cm x 0.46cm i.d., 5µm particle size) was used to establish an isocratic RP-HPLC on a Shimadzu HPLC system. The mobile phase contained a mixture of buffer, Acetonitrile 35:65; the buffer preparation was prepared weighed 5.23g of potassium phosphate dibasic in 1000 ml of water, stirred well, and the pH was adjusted to 7.0 using orthophosphoric Acid (phosphoric Acid, H3PO4). A mobile phase was made (35:65), with Acetonitrile at 35 volumes and buffer at 65 volumes. The peak elution was observed at 220nm, with a 1.2ml/min flow rate; Impurity A 3.89 minutes, impurity B 32.85 minutes, and MH 20.69 minutes were determined to have retention times, respectively. The calibration curve used to assess MH and its related impurities (Impurity A: 10-(3-(dimethylamino) propyl)-9,10-dihydro-9,9-dimethylanthracen-10-ol, and Impurity B: 10,10-dimethyl anthrone), demonstrates a straight line across the concentration range of 5µg/ml to 200µg/ml, with an r2 value of 0.999. The limit of quantification is 29.68µg/ml for MH, 7.95µg/ml for Impurity A, and 17.08µg/ml for Impurity B. Statistical analysis has been conducted on the outcomes obtained from our proposed methodologies. The API drugs were subjected to Acid, base, thermal, UV, sunlight, and peroxide degradation. The degradation studies indicated MH showed degradation only in peroxide conditions. With notable variations in their retention time values, the degradation products from the pure medication were clearly separated. Melitracen hydrochloride can be quantitatively analyzed simultaneously in bulk medications with success when using this approach.
Cite this article:
Bhaskar Mangalapu, B. Srinivas, Sirigiri Ramathulasi, Ramesha Andagar Ramakrishna, Venkata Narayana Palakollu. Stability Indicating Method Development and Validation for Quantification of Impurities in Melitracen Hydrochloride, An Antidepressant Drug Using Reverse Phase High-Performance Liquid Chromatography (RP-HPLC). Research Journal Pharmacy and Technology. 2025;18(8):3730-8. doi: 10.52711/0974-360X.2025.00537
Cite(Electronic):
Bhaskar Mangalapu, B. Srinivas, Sirigiri Ramathulasi, Ramesha Andagar Ramakrishna, Venkata Narayana Palakollu. Stability Indicating Method Development and Validation for Quantification of Impurities in Melitracen Hydrochloride, An Antidepressant Drug Using Reverse Phase High-Performance Liquid Chromatography (RP-HPLC). Research Journal Pharmacy and Technology. 2025;18(8):3730-8. doi: 10.52711/0974-360X.2025.00537 Available on: https://www.rjptonline.org/AbstractView.aspx?PID=2025-18-8-40
REFERENCES:
1. Tripathi KD. Essentials of medical pharmacology. Jaypee Brothers Medical Publishers. 2018; Oct 31.
2. O'Neil MJ, editor. The Merck index: an encyclopedia of chemicals, drugs, and biologicals. RSC Publishing. 2013.
3. Gatti R, Andreatta P, Gioia MG, Boschetti S. An optimized and validated reversed-phase chromatographic method for the simultaneous determination of glucosamine and chondroitin sulphate in dietary supplements. InXIX National Meeting on Medicinal Chemistry. Book of Abstracts 2008 (pp. 250-250). ITA.
4. Uddin M, Mamun A, Rashid M, Asaduzzaman M. In-process and finished products quality control tests for pharmaceutical capsules according to pharmacopoeias. British Journal of Pharmaceutical Research. 2016; Jan 10; 9(2): 1-9. https://doi.org/10.9734/BJPR/2016/22044
5. Toronto O. The Official Compendia of Standards, Webcom Limited; 2005. The United States Pharmacopoeia-National Formulary.
6. Keitel S. Pharmacopoeial Standards: European Pharmacopoeia. Encyclopedia of Pharmaceutical Science and Technology,. 2013 Jul 1:2691-703.
7. Chhalotiya UK, Bhatt KK, Shah DA, Nagda CD, Patel JR. Development OF HPTLC Method For The Estimation Of Antidepressant Drugs Melitracen And Flupentixol In Their Combined Dosage Form. Journal of Liquid Chromatography and Related Technologies. 2013; Mar 1; 36(9): 1231-42. https://doi.org/10.1080/10826076.2012.685920
8. Sharma MC, Sharma S, Sharma AD. Novel application and spectrophotometric estimation of Melitracen HCl tablet dosage form using niacinamide as hydrotropic solubilizing agent. Journal of Chemical and Pharmaceutical Research. 2010; 2(2): 416-20.
9. Sheikh IA, Charde M, Kasture AV. Development of Spectrophotometric Method for Simultaneous Estimation of Flupenthixol HCl and Melitracen HCl in Their Combined Dosage Form. Asian Journal of research in Chemistry. 2009; 2(4): 488-93.
10. Xu P, Li HD, Zhu YG, Chen BM, Ma N, Xie YL, Zhang BK. Validated liquid–liquid extraction and LC–ESI–MS method for the determination of melitracen in human plasma. Chromatographia. 2008; Jun; 67: 935-9. https://doi.org/10.1365/s10337-008-0619-1
11. Tanaka E, Terada M, Nakamura T, Misawa S, Wakasugi C. Forensic analysis of eleven cyclic antidepressants in human biological samples using a new reversed-phase chromatographic column of 2 μm porous microspherical silica gel. Journal of Chromatography B: Biomedical Sciences and Applications. 1997; May 9; 692(2): 405-12. https://doi.org/10.1016/S0378-4347(96)00510-5
12. Kollroser M, Schober C. Simultaneous determination of seven tricyclic antidepressant drugs in human plasma by direct-injection HPLC-APCI-MS–MS with an ion trap detector. Therapeutic drug monitoring. 2002; Aug 1; 24(4): 537-44.
13. Shinozuka T, Terada M, Tanaka E. Solid-phase extraction and analysis of 20 antidepressant drugs in human plasma by LC/MS with SSI method. Forensic science international. 2006; Oct 16; 162(1-3): 108-12. https://doi.org/10.1016/j.forsciint.2006.03.038
14. Che J, Meng Q, Chen Z, San C, Hou Y, Cheng Y. Validation of a sensitive LC/MS/MS method for simultaneous quantitation of flupentixol and melitracen in human plasma. Journal of pharmaceutical and biomedical analysis. 2007; Dec 21; 45(5): 785-92. https://doi.org/10.1016/j.jpba.2007.07.022
15. Chhalotiya UK, Bhatt KK, Shah DA, Nagda CD, Patel JR. Development of Hptlc Method For The Estimation Of Antidepressant Drugs Melitracen And Flupentixol In Their Combined Dosage Form. Journal of Liquid Chromatography and Related Technologies. 2013; Mar 1; 36(9): 1231-42. https://doi.org/10.1080/10826076.2012.685920
16. ICH I. Q2 (R1): Validation of analytical procedures: text and methodology. InInternational conference on harmonization, Geneva 2005 Nov 6 (Vol. 2005).
17. ICH, Technical Requirements for the Registration of Pharmaceutical for Human Use; Validation of Analytical Procedures: Text and Methodology Q2 (R1); IFPMA: Geneva, Switzerland, 2005.
18. ICH I. Q1B, Stability testing: photostability testing of new drug substances and products. InInternational Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use, Geneva 1996.