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BR-102024017737-A2 - PROCESS FOR DETECTION AND ANALYSIS OF CISPLATIN AND RELATED SUBSTANCES BY MEANS OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)

BR102024017737A2BR 102024017737 A2BR102024017737 A2BR 102024017737A2BR-102024017737-A2

Abstract

The present invention describes an analytical process, based on the use of HPLC, that allows the identification and quantification of the drug cisplatin in aqueous solutions, pharmaceutical and biological samples such as blood serum and body fluids, even at low concentrations (parts per billion).

Inventors

  • DANIEL CRISTIAN FERREIRA SOARES
  • MATHEUS VALENTIN MAIA
  • DANIEL BRAGANÇA VIANA
  • STEPHANIE CALAZANS DOMINGUES

Assignees

  • UNIFEI - UNIVERSIDADE FEDERAL DE ITAJUBÁ

Dates

Publication Date
20260310
Application Date
20240828

Claims (8)

  1. 1. Process for the detection and analysis of cisplatin using High-Performance Liquid Chromatography (HPLC), characterized by comprising the following steps: a. Subjecting the sample to liquid chromatography in a liquid chromatography column coupled to a diode array detector (DAD), where the stationary phase used is a C18 reversed-phase column. b. Eluting the sample using a mobile phase, with a minimum flow rate of 0.1 mL/min up to 3.0 mL/min at a controlled temperature, preferably employing a flow rate of 1.5 mL/min at 27°C. c. Identifying and quantifying cisplatin or related substances in samples by means of detection in the ultraviolet spectrum region (200 to 600 nm, preferably at 210 nm) and developing a calibration curve. d. Quantifying cisplatin in the sample using a calibration curve.
  2. 2. Process for detection and quantification of cisplatin in aqueous, pharmaceutical and biological samples, according to claim 1, step “a”, characterized by having the stationary phase consisting of a C18 reversed-phase column.
  3. 3. Method for detection and quantification of cisplatin in aqueous, pharmaceutical and biological samples, according to claim 1, step “b”, characterized by having the mobile phase composed of a mixture of methanol and water.
  4. 4. Method for detection and quantification of cisplatin in aqueous, pharmaceutical and biological samples according to claim 3, characterized by the ratio of deionized water (25%) and methanol (75%).
  5. 5. Use of the process defined by claims 1 to 4, characterized by being used for the identification or determination of cisplatin during the development of pharmaceutical formulations based on the use of cisplatin for use in the treatment of cancer.
  6. 6. Use of the process defined by claims 1 to 4, characterized by the identification or determination of cisplatin during the quality control of the production of injectable solutions.
  7. 7. Use of the process defined by claims 1 to 4, characterized by the identification or determination of cisplatin in samples of hospital waste and water and sewage treatment plants to investigate and control environmental contamination.
  8. 8. Use of the process defined by claims 1 to 4, characterized by the identification and determination of cisplatin in various biological samples.

Description

FIELD OF THE INVENTION: [001] The present invention is situated in the area of analytical chemistry and/or pharmaceutical sciences, and is characterized by an innovative process that allows the quantification of the drug cisplatin in aqueous medium through the technique of high-performance liquid chromatography. BACKGROUND OF THE INVENTION / RELATED TECHNIQUE: [002] Cisplatin is one of the most widely used chemotherapeutic agents worldwide for cancer treatment, a fact associated with its broad spectrum of therapeutic efficacy against many types of solid tumors. It is classified as an alkylating agent, chemically consisting of a platinum atom bonded to two chlorine atoms and two ammonium groups in the cis spatial position. In use since 1978, when it was approved by the FDA, the drug has been primarily employed in the treatment of tumors in the ovary, bladder, testicle, and lung. [003] Disadvantages such as the significant nephrotoxicity, cardiotoxicity, nausea, and vomiting caused by antitumor drugs in their free form have motivated the development of controlled-release pharmaceutical formulations such as PEGylated liposomes (Caelyx®) and non-PEGylated liposomes (Myocet®), which received authorization for use from the American FDA in 1996 and 2000, respectively. These new formulations have significantly improved the toxicological profile of the drug; however, with prices still very high today, access to the medication is restricted to a small portion of the population. Formulations containing cisplatin in its free form still comprise almost all pharmaceutical formulations currently in clinical use in Brazil and many countries worldwide. [004] Cisplatin exhibits absorptivity in the UV-VIS region, which can be enhanced by derivatization methods; however, it does not fluoresce. The main analytical methods currently used for quantifying the drug are based on the use of UV-VIS spectroscopy techniques and high-performance liquid chromatography (HPLC). There are published works in the literature employing HPLC analysis protocols for cisplatin with a UV-VIS detector; however, works describing protocols for high-performance liquid chromatography using a diode array detector (HPLC-DAD) are scarce and do not include validation steps, a necessary process for laboratory studies, where it is evaluated whether the performance of the proposed method and the established parameters meet the requirements for the intended analytical application. In this sense, the present invention is based on a set of innovative tasks that allow the dosage of cisplatin via HPLC technique using DAD that follows the standards of RDC No. 166 of the National Health Surveillance Agency (ANVISA). BRIEF DESCRIPTION OF THE FIGURES: [006] Figure 1- Cisplatin chromatograms obtained at a concentration of 50μg mL-1 by HPLC. [007] Figure 2 - Analytical calibration curve of cisplatin obtained by HPLC technique in the concentration range of 20-100 μg mL-1 [008] Figures 3 and 4 - Study of the selectivity of the method for detection and quantification of cisplatin sample in phosphate and ferric chloride buffer. DETAILED DESCRIPTION OF THE INVENTION: [009] The present invention describes a process based on HPLC technique for detecting and quantifying the drug cisplatin in aqueous pharmaceutical and biological samples, such as pharmaceutical formulations, prepared drug solutions and blood serum. [010] The process for detecting and quantifying cisplatin in pharmaceutical and biological samples comprises the following steps: a. Subjecting the sample to HPLC in a C18 reverse liquid chromatography column, where the stationary phase used in the column is characterized by C18 silica particles. b. Eluting the sample using a mobile phase, with a minimum flow rate of 0.1 mL/min up to 3.0 mL/min, at a controlled temperature, preferably using a flow rate of 1.5 mL/min at 25°C; c. Identifying and quantifying cisplatin or related substances in samples by means of detection in the ultraviolet spectrum region (200 to 600 nm, preferably at 210 nm) and developing a calibration curve. [011] In step “b”, the mobile phase may consist of an organic solvent which may comprise mixtures of the following solvents: methanol and water. The solvent-to-water ratio shall comprise the proportion of deionized water (25%) and methanol (75%). [012] The proposed process can be used for the determination of cisplatin during the development of cisplatin-based pharmaceutical formulations for use in cancer treatment; determination of cisplatin during quality control of injectable solutions production; determination of cisplatin in samples of hospital waste and water and sewage treatment plants to investigate and control environmental contamination resulting from its use; determination of cisplatin in various biological samples. [013] US patent 6224883B1 describes a process for preparing cisplatin suspensions with greater precision and homogeneity by employing a diluent containing a non-ionic and bioc