High-performance liquid chromatographic determination of the novel antitumour drug topotecan and topotecan as the total of the lactone plus carboxylate forms, in human plasma

doi:10.1016/0378-4347(95)00054-M

Journal of Chromatography B: Biomedical Sciences and Applications

Volume 668, Issue 1, 9 June 1995, Pages 107-115

https://doi.org/10.1016/0378-4347(95)00054-M Get rights and content

Abstract

A sensitive high-performance liquid chromatographic (HPLC) assay has been developed and validated for the quantitation of the novel anticancer agent topotecan and topotecan as the total of its lactone and carboxylate forms in human plasma. Linear response in analyte standard peak area were observed over the concentration range 0.05–10 ng/ml using 100-μl plasma samples. The instability of the drug in the biological matrix necessitated that the plasma fraction was obtained within 5 min after blood sampling by centrifugation, immediately followed by protein precipitation with cold methanol (−30°C). Stability studies have indicated that topotecan is stable in these methanolic extracts for at least 4.5 months at −30°C and 2 months at −70°C. For the total determination of the lactone plus lactone ring-opened forms of the drug as topotecan, plasma samples were deproteinated with methanol and, subsequently, acidified with 7% (v/v) perchloric acid. Plasma samples for the measurement of total levels of the lactone and the ring-opened forms of the topotecan were stable for at least 4.5 months when stored at −30°C. After centrifugation, the supernatants were analysed by HPLC using a Zorbax SB-C₁₈ Stable Bond column and methanol-0.1 M hexane-1-sulfonic acid in methanol-0.01 M N,N,N′,N′-tetramethylethylenediamine (TEMED) in distilled water pH 6.0 (25:10:65, v/v) as the mobile phase. Detection was performed fluorimetrically. Within-run and between-run precision was always less than 12.1% in the concentration range of interest (0.05–10.0 ng/ml). The limit of quantitation is 0.05 ng/ml. Accuracy measurements ranged between 87.6 and 113.5%.

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Cited by (92)

Oral delivery of topotecan in polymeric nanoparticles: Lymphatic distribution and pharmacokinetics
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Citation Excerpt :
Lactone and carboxylated forms differ in their pharmacological activities. Specifically, the lactone form has higher anti-cancer activity than the carboxylated form [12]. Chemical structures of both lactone and carboxylated forms of topotecan are shown in Fig. 1.
There have been many attempts to formulate a variety of drugs in nano-size formulations. However, biodistribution characteristics of these formulated drugs remain unclear. Information about the pharmacokinetics and distributions of these formulations is essential for future practical use and advanced formulation development. Topotecan is a useful agent for treating a variety of cancers. It exhibits anti-cancer activity by inhibiting topoisomerase. However, oral bioavailability of topotecan was not satisfactory in previous studies. Reversible hydrolysis of its active site according to pH environment was a major limitation in terms of treatment. To improve the bioavailability and retention of topotecan in target organs (such as lung and brain) and increase its delivery to the lymphatic system as a major pathway for cancer metastasis, this study was conducted on topotecan-loaded nanoparticles using poly(lactic-co-glycolic acid) (PLGA). These nanoparticles were prepared by double emulsion solvent evaporation. Formulated topotecan-loaded PLGA nanoparticles were subjected to several in vitro tests to determine various physicochemical properties such as size, zeta potential, encapsulation efficiency, morphology, and release profile. These nanoparticles were also subjected to in vivo studies using rats. Based on in vivo results, pharmacokinetic properties, distribution in the body, and delivery efficiency of these formulated nanoparticles were confirmed. Topotecan-loaded PLGA nanoparticles showed a delayed release pattern in vitro. Their pharmacokinetic profiles and distributions in the body were clearly different from those of free topotecan hydrochloride. Results confirmed that topotecan encapsulated in the PLGA polymer was stable from hydrolysis and present in an active form for a longer time in the body. Biometric imaging revealed in vivo properties of topotecan-loaded PLGA nanoparticles for qualitative confirmation. And oral delivery of topotecan in polymeric nanoparticles to lymph and various body tissues has been identified. Findings of this study indicate that topotecan formulated into nanoparticles (using PLGA) has a better pharmacokinetic profile and a better delivery to lymphoid tissues, lung, and brain than free topotecan hydrochloride, suggesting that these topotecan-loaded PLGA nanoparticles might provide better therapeutic results.
A novel electrochemical sensing platform based on mono-dispersed gold nanorods modified graphene for the sensitive determination of topotecan
2020, Sensors and Actuators, B: Chemical
Citation Excerpt :
Antineoplastic agents play a key role in the treatment of various cancer types with many side effects for human body. Topotecan (TPC), a water-soluble semi-synthetic comptothecin derivative, has been extensively used in the treatment of certain types of cancer such as breast, lung and ovarian [1,2]. TPC selectively inhibits the topoisomerase I activity, causing a damage on DNA strands and consequently a death of cancer cells [3].
In this research, we developed a novel design to fabricate a sensitive and selective electrochemical sensor based on high-quality graphene decorated with mono-dispersed gold nanorods (AuNRDs) for the determination of topotecan (TPC) in human blood serum. For this purpose, the screen-printed electrode (SPE) was firstly modified with graphene prepared by metal intercalation process (MI-GR), and then followed by the decoration of mono-dispersed AuNRDs synthesized by seed mediated growth method. The synthesized MI-GR and AuNRDs samples were characterized by Raman, XPS, UV-Vis and TEM techniques. Electrochemical studies indicated that the electrocatalytic activity of MI-GR has superior than that of chemically reduced graphene (CR-GR) prepared by conventional approach due to its faster electron transfer ability. Moreover, AuNRDs/MI-GR modified SPE (AuNRDs/MI-GR/SPE) demonstrated a wide linearity in the range of 0.1 – 16.0 μM with a low detection limit of 22.0 nM. The analytical reliability of AuNRDs/MI-GR/SPE was successfully confirmed by the detection of TPC in human blood serum with the satisfactory recoveries at the range of 99.8–100.07%. AuNRDs/MI-GR/SPE could be utilized as an alternative analytical tool for the determination of anti-cancer agents such as TPC in the next generation clinical applications.
Synergic effect of 2D nitrogen doped reduced graphene nano-sheet and ionic liquid as a new approach for fabrication of anticancer drug sensor in analysis of doxorubicin and topotecan
2018, Journal of Molecular Liquids
Citation Excerpt :
Although doxorubicin showed a powerful ability in chemotherapy process, its high side effects, such as hair loss, nausea and immunosuppression are still a major challenge and the main problem; thus determines of its level in the human body is necessary for a chemotherapy process [41]. Topotecan or hycamtin (as a topoisomerase inhibitor) is suggest as a powerful the anticancer drug for treat certain cases of lung cancer, breast cancer and ovarian cancer [42]. The asthenia, vomiting, myelosuppression, and thrombocytopenia are the major side effects of topotcan after chemotherapy process.
The determination of doxorubicin and topotecan as two important breast anticancer drugs with high side effects is significant in pharmaceutical samples. 2D nitrogen doped reduced graphene (NrG) nano-sheet and 1-methyl-3-octylimidazolium chloride (MOICl) were suggested as two high conductive modifiers for fabrication of anticancer drug sensor based carbon paste electrode (CPE). The electrical conductivity of NrG/MOICl/CPE as a new analytical approach was probed by voltammetric methods. The NrG/MOICl/CPE showed accelerating the exchange of electrons for oxidation of doxorubicin and this improving helps to simultaneous determination of doxorubicin and topotecan with separation gap of 315 mV. The NrG/MOICl/CPE as an anticancer sensor displayed detection limits of 3.1 nM and 0.27 μM for doxorubicin and topotecan, respectively. The NrG/MOICl/CPE was found to be practically applicable in the determination of doxorubicin and topotecan in drug samples.
Electrochemical detection of anticancer drug topotecan using nano-acetylene black film
2011, Colloids and Surfaces B: Biointerfaces
Citation Excerpt :
So, the quantitative detection of topotecan is quite important in the clinic. Up-to-now, several methods have been reported for the detection of topotecan, such as high-performance liquid chromatography (HPLC) [3–6], liquid chromatography–mass spectrometry (LC–MS) [7] and fluorescence [8]. From Fig. 1, it is known that topotecan contains a phenolic hydroxyl group, which is electrochemical active and can be oxidized under proper conditions.
Topotecan, a novel anticancer drug, has been widely used in the treatment of ovarian and cervical cancers. Herein, acetylene black (AB) nanoparticles were used to modify the electrode surface, and the electrochemical behavior of topotecan was examined. At the AB film surface, an irreversible oxidation peak was observed for topotecan, and the response signal greatly increased. The influences of pH value, amount of AB, accumulation potential and time were investigated. As a result, a novel electrochemical method with high sensitivity and rapid response was developed for the detection of topotecan. The linear range is from 2 μg L⁻¹ to 0.4 mg L⁻¹, and the limit of detection is 1.45 μg L⁻¹ (or 3.17 × 10⁻⁹ M). The method was successfully used to detect topotecan in blood serum, and the results consisted with the values that obtained by high performance liquid chromatography.
Concurrent determination of topotecan and model permeability markers (atenolol, antipyrine, propranolol and furosemide) by reversed phase liquid chromatography: Utility in Caco-2 intestinal absorption studies
2007, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
A simple, sensitive, specific and high-resolution reversed-phase liquid chromatographic method utilizing ultraviolet detection has been developed and validated for simultaneous determination of topotecan and four intestinal permeability markers (atenolol, antipyrine, propranolol and furosemide) as suggested by US-FDA. Chromatography was carried out on C-18 column with mobile phase comprising water (pH 3.0) and acetonitrile gradient pumped at a flow rate of 1 ml min⁻¹. The validation parameters included specificity, accuracy, precision, sensitivity and stability studies. Topotecan, an anti-cancer drug widely used in metastatic carcinoma, is a P-glycoprotein substrate having oral bioavailability of 30% with large inter-patient variability. The present method was successfully applied for demonstrating P-gp mediated transport of topotecan and its inhibition using verapamil in Caco-2 cell monolayer. The method can be used in identification of novel P-gp inhibitors for topotecan and estimating the contribution of P-gp in affecting oral bioavailability of topotecan. The other applications of method include its use in validation of Caco-2 monolayer assay for getting biowaiver based on Biopharmaceutic Classification System and its extrapolation to in situ and/or in vivo studies.
Robust Inclusion Complex of Topotecan Comprised within a Rhodamine-Labeled β-Cyclodextrin: Competing Proton and Energy Transfer Processes
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Regular paperHigh-performance liquid chromatographic determination of the novel antitumour drug topotecan and topotecan as the total of the lactone plus carboxylate forms, in human plasma

Abstract

Ann. Oncol.

Ann. Oncol.

Cancer Treat. Rev.

J. Pharm. Biomed. Anal.

J. Pharm. Sci.

J. Pharm. Biomed. Anal.

J. Chromatogr.

J. Chromatogr.

J. Chromatogr.

J. Pharm. Sci.

J. Med. Chem.

Regular paper
High-performance liquid chromatographic determination of the novel antitumour drug topotecan and topotecan as the total of the lactone plus carboxylate forms, in human plasma