Determination Of Irinotecan Research Articles

A Novel Technique for the Quantification of Irinotecan HCl Injection

This work aimed for the method development and validation of RP-HPLC method for quantitative determination of Irinotecan HCL in bulk and pharmaceutical formulation. More precise, accurate, sensitive, simple and economic HPLC method was developed and validated for quantitative determination of Irinotecan in bulk and dosage forms. Irinotecan was analyzed by using reverse phase Inertsil ODS 250 X 4.6 mm and 5μ particle size column. 0.02 M KH2PO4 buffer and acetonitrile were used as a mobile phase in the ratio of 40: 60 v/v. The pH of the buffer was adjusted to 3.2 with o-phosphoric acid. The flow rate was 1.0 ml/min and the run time was 5 min. The analysis was carried out at 222 nm with UV detector at ambient temperature. The developed HPLC method was validated and stability studies were conducted under different conditions. Irinotecan was eluted at retention time 2.1 min and the calibration curve was linear in the range 40–120 μg/ml concentration. Correlation coefficient was found to be 0.9999. The Irinotecan test and standard stock solutions were stable over the period of 5 days. The LOD and LOQ were found to be 0.8 ng/ml and 2.0 ng/ml. % RSD of Irinotecan content was < 0.5% and the assay was found to be 98.2–100%. The proposed method is observed to be rapid and selective, when compared to the methods reported in the literature. The retention time of Irinotecan was found to be very less (2.1 min). The method is most precise and economic with respect to solvent consumption.

Determination of irinotecan and its metabolite SN-38 in dried blood spots using high-performance liquid-chromatography with fluorescence detection

Irinotecan (IRI) is an antineoplastic drug widely used for the treatment of colorectal and advanced pancreatic cancer. Despite its clinical utility, the clinical use of IRI is associated with potentially severe hematopoietic and gastrointestinal toxicities. The quantification of IRI and its active metabolite SN-38 in dried blood spots (DBS) may be an alternative to individualize the drug dose through a minimally invasive and easy collection method. The aim of this study was to develop and validate a simple and fast HPLC-FL assay for simultaneous IRI and SN-38 measurement in DBS, with adequate analytical performance for clinical use. The method employs liquid extraction of one 8mm disk of whole blood, followed by separation in a reversed phase Eclipse Plus C8 column (150×4.6mm, 5μm). Detection was performed with a fluorescence detector, with excitation wavelength of 370 and emission of 420 for IRI and 540nm for SN-38 and internal standard (camptothecin). Total analytical run time was 17min. Mobile phase was a mixture of 0.1M phosphate buffer pH 4.0 and acetonitrile (80:20, v/v), at 1mLmin−1. The assay was linear in the range 10–3,000ngmL−1 and from 0.5 to 300ngmL−1 for IRI and SN-38, respectively. Precision assays presented CV% of 2.71-5.65 and 2.15-10.07 for IRI and SN-38, respectively, and accuracy in the range of 94.26-100.93 and 94.24-99.33%. IRI and SN-38 were stable at 25 and 42°C for 14days in DBS samples. The method was applied to DBS samples obtained from fingerpicks from 19 volunteers receiving IRI in single or combined chemotherapy regimens, collected 1 and 24h after beginning of the infusion. The estimated plasma concentration of IRI and SN-38 in sample collected 1h after star of infusion had 16 of 19 values within the ±20% range of the measured plasma concentrations. On the other hand, predictions of IRI and SN-38 plasma concentrations from DBS measurements obtained 24h after the beginning of the infusion were poor. AUC of IRI that was calculated using plasma and DBS-estimated concentrations, with a high correlation (r=0.918). The method presented suitable characteristics for the clinical use. However, translation of IRI and SN-38 DBS to plasma concentrations is challenging due to the compound’s variable plasma/blood partition.

Determination of irinotecan, SN‐38 and SN‐38 glucuronide using HPLC/MS/MS: Application in a clinical pharmacokinetic and personalized medicine in colorectal cancer patients

Irinotecan (CPT-11) is chemotherapy used mainly in the metastatic colorectal cancer. The purpose of this study was to develop and validate the LC-MS/MS for the simultaneous determination of CPT-11, SN-38, and SN-38G. A 100μL of plasma was prepared after protein precipitation and analyzed on a C18 column using 0.1% acetic acid in water and 0.1% acetic acid in acetonitrile as mobile phases. The mass spectrometer worked with multiple reaction monitoring (MRM) in positive scan mode. The standard curves were linear on a concentration range of 5-10000ng/mL for CPT-11, 5-1000ng/mL for SN-38, and 8-1000ng/mL for SN-38G. In this assay, the intra and interday precision consisted of ≤9.11% and ≤11.29% for CPT-11, ≤8.70% and 8.31% for SN-38, and ≤9.90 and 9.64% for SN-38G. This method was successfully used to quantify CPT-11, SN-38, and SN-38G and applied to a pharmacokinetic study.

Development of LC–MS/MS Method for Simultaneous Determination of Irinotecan and Its Main Metabolites in Rat Plasma, and Its Application in Pharmacokinetic Studies

The anticancer agent irinotecan (CPT-11) is widely used in several cancer regimens. However, drug–drug interaction effects and considerable interindividual variability in the pharmacokinetics of CPT-11 and its metabolites have been observed, leading to side effects. To solve these problems and promote CPT-11 treatment, we established and validated a simple, rapid, and sensitive LC–MS/MS method for the simultaneous quantitative determination of CPT-11 and its main metabolites (SN-38, SN-38G, and APC). Protein precipitation with methanol was selected for sample preparation. All separations were performed on an Agilent ZORBAX Eclipse XDB-C18 column (2.1 × 50 mm, 3.5 μm) under isocratic elution with a mobile phase consisting of acetonitrile and 0.1% formic acid. The mass spectrometer was operated with multiple reaction monitoring in positive ion mode. The method was validated as linear over the concentration ranges 16.9–6760 ng mL−1 for CPT-11, 2.3–920 ng mL−1 for SN-38, 2.5–1000 ng mL−1 for SN-38G, and 1.25–250 ng mL−1 for APC. The intra- and interbatch precisions and accuracies of the validated method were within acceptable limits (<15%). This method was successfully applied to a pharmacokinetic study of CPT-11 and its main metabolites in rats.

Ionic Liquid Dispersive LiquidLiquid Microextraction Method for the Determination of Irinotecan, an Anticancer Drug, in Water and Urine Samples Using UV-Vis Spectrophotometry.

A new, simple, efficient, and environmentally friendly ionic liquid dispersive liquid&amp;#x2013;liquid microextractionmethod was developed for the determination of irinotecan, an anticancer drug, in water and urine samples using UV-Vis spectrophotometry. The ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate was used as the extraction solvent, and ethanol was used as the disperser solvent. The main parameters affecting the extraction efficiency, including sample pH, volume of the ionic liquid, choice of the dispersive solvent and its volume, concentration of NaCl, and extraction and centrifugation times, were investigated and optimized. The effect of interfering species on the recovery of irinotecan was also examined. Under optimal conditions, the LOD (3&amp;#x03C3;) was 48.7 &amp;#x03BC;g/L without any preconcentration. Because the urine sample was diluted 10-fold, the LOD for urine would be 487 &amp;#x03BC;g/L. However, this could be improved 16-fold if preconcentration using a 40 mL aliquot of the sample is used. The proposed method was successfully applied to the determination of irinotecan in tap water, river water, and urine samples spiked with 10.20 mg/L for the water samples and 8.32 mg/L for the urine sample. The average recovery values of irinotecan determined were 99.1% for tap water, 109.4% for river water, and 96.1% for urine.

Determination of irinotecan and SN38 in human plasma by TurboFlow™ liquid chromatography–tandem mass spectrometry

Irinotecan is a cytotoxic agent used in the treatment of metastatic colorectal cancer. Irinotecan is a prodrug when is converted in vivo to an active metabolite SN38, which has potent pharmacological activity. SN38 is then inactivated and excreted as SN38-glucuronide. High-performance liquid chromatography–mass spectrometry is a widely used bioanalysis technique that can be coupled to the turbulent-flow extraction line to shorten preparation time. A technique was developed to quantify irinotecan and its metabolite by liquid chromatography–tandem mass spectrometry coupled with a turbulent-flow online extraction method.Assays were performed on 100μL of plasma after protein precipitation. The supernatant is injected directly into the extraction column, transferred to the chromatographic column, and analyzed by tandem mass spectrometry.Linearity, reproducibility and repeatability of the method were validated on a concentration range of 25–2500ng/mL for irinotecan and 5–500ng/mL for SN38. For the low limit of quantification of irinotecan and SN38, precision is 6.31% and 8.73%, and accuracy is 84.0% and 91.8%, respectively. The SN38-glucuronide determination protocol included a hydrolyzation step.This method was successfully used to quantify irinotecan, SN38 and SN38-G in human plasma in a clinical trial.

Determination of irinotecan and its metabolite SN-38 in rabbit plasma and tumors using a validated method of tandem mass spectrometry coupled with liquid chromatography.

New tandem mass spectrometric method coupled with liquid chromatography (LC-MS/MS) has been developed to determine the total concentration of camptothecin derivatives (irinotecan and SN-38) regardless of inter-conversion phenomenon between carboxylate and lactone forms. At first, all sample solutions were acidified for 1h in order to completely convert CPT derivatives into their lactone forms and then CPT derivatives were extracted with organic solution containing diethyl ether and ethyl acetate (2:1, v/v) just after alkalization in the range pH 8.0-8.5 in acid-treated solutions. Analytes were separated on a reverse phase C18 column (150×2.1mm) and eluted isocratically with a mobile phase which consisted of acetonitrile-methanol-buffer (0.1% formic acid, 5mM ammonium formate) (3:4:3, v/v). CPT derivatives were monitored by tandem mass spectrometry in electrospay-positive ionization and multiple reaction mode programmed to the following transitions (m/z): '587.6→167.2' of CPT-11, '393.6→349.3' of SN-38 and '349.4→ 305.2' of CPT. The method was validated to have the proper linearity (r(2)>0.99) over the range of 5-1000ng/ml of CPT-11 and 1-250ng/ml of SN-38 with good accuracy (89.8-114.3%) and precision (less than 10%). In all stability tests, concentration of CPT-11 and SN-38 had been left in the acceptable range of 88.8-110.7% when sample solutions were acidified before determination of CPT derivatives. Newly developed LC-MS/MS method was suitable for the determination of CPT derivatives of both rabbit plasma and tumor tissues in the pharmacokinetic study.

SIMULTANEOUS DETERMINATION OF DOXORUBICIN AND IRINOTECAN IN CONJUNCTION WITH THEIR MAJOR METABOLITES BY ULTRA HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

An accurate UPLC method was developed and validated for the simultaneous determination of irinotecan and doxorubicin extracted from murine plasma, along with their major metabolites SN-38 and doxorubicinol, respectively. The parent drugs and metabolites were extracted from plasma by protein precipitation method. Camptothecin was used as the internal standard. Chromatographic separation was performed using an ACQUITY UPLC BEH C18 column (150 × 2.1 mm, particle size of 1.7 µm) at 40°C by means of gradient elution. The mobile phases for gradient elution were consisted of 0.1 % v/v formic acid in either acetonitrile or water. The flow rate was 0.5 mL/min, and the total run time was 7 min. The linear quantitation ranges for parent drugs (irinotecan and doxorubicin) and for metabolites (doxorubicinol and SN-38) were 250–10000 ng/mL and 125–5000 ng/mL, respectively (r2 > 0.99). The lower limit of quantification (LLOQ) for irinotecan and doxorubicin was 250 ng/mL, and the LLOQ for SN-38 and doxorubicinol was 125 ng/mL. The intra- and inter-day relative standard deviation was <10 %, and accuracy was >90%.

Simultaneous Determination of Irinotecan and Its Metabolite 7-Ethyl-10-hydroxycamptothecin in Biological Fluids Using Excitation-emission Matrix Fluorescence Coupled with Second-order Calibration Method

提出了激发发射矩阵荧光光谱与化学计量学二阶校正方法相结合用于同时快速定量人体液（血浆和尿液）中的伊立替康（CPT11）和其主要代谢产物7-乙基-10-羟基喜树碱（SN38）的绿色、高灵敏分析策略.尽管其分析物之间以及分析物和背景之间的光谱存在严重重叠现象,采用基于交替归一加权残差（ANWE）算法的二阶校正方法进行解析仍能得到令人满意的定性定量分析结果.当该体系的组分数选取为3时,可以得到血浆和尿液中CPT11的平均回收率分别为（96.8±6.3）%和（101.7±1.1）%,SN38在血浆和尿液中的平均回收率分别为（100.4±4.9）%和（101.6±1.1）%.另外,通过品质因子,如灵敏度（SEN）、选择性（SEL）、检测下限（LOD）和定量检测限（LOQ）评估了该方法的准确性.实验结果表明,该方法能以＂数学分离＂代替繁琐的＂物理和化学分离＂,成功地解决实际复杂体系中内源干扰物质与分析物光谱重叠所引起的难分辨的问题,可用于人体液中CPT11和SN38含量的直接快速定量测定.

An Accurate Assay for Simultaneous Determination of Irinotecan and Its Active Metabolite SN-38 in Rat Plasma by LC with Fluorescence Detection

An accurate LC method was developed and validated for simultaneous determination of irinotecan (CPT-11) and its active metabolite SN-38 in rat plasma. Plasma samples were pretreated with 0.4 g mL−1 sodium dodecyl sulfate to inactive the carboxylesterase and avoid the conversion of CPT-11 to SN-38. Chromatographic separation was achieved on a Diamaonsil C18 column using acetonitrile–50 mM phosphate buffered solution (30:70, v/v) at pH 4.0 as the mobile phase with the flow rate of 1 mL min−1. The linear quantitation ranges for CPT-11 and SN-38 were 5.05–3,030 and 3.15–315 ng mL−1 with r 2 > 0.99, respectively. The lower limit of quantification (LLOQ) was 2.33 ng mL−1 for CPT-11 and 0.26 ng mL−1 for SN-38 with intra- and inter-day relative standard deviation of 90%. The method was proved to be accurate and sensitive enough and was successfully applied to a pharmacokinetic study of CPT-11 in rats.

Determination of irinotecan (CPT-11) and SN-38 in human whole blood and red blood cells by liquid chromatography with fluorescence detection

An analytical method was developed for the anticancer agent irinotecan (CPT-11) and its main metabolite SN-38 in human whole blood and in red blood cells (RBCs). Sample pretreatment involved deproteinization of whole blood or plasma-diluted RBCs isolated by MESED instruments, with a mixture of aqueous perchloric acid and methanol (1:1, v/v). Separation was carried out using isocratic elution on a Hypersil ODS stationary phase, with detection at excitation and emission wavelengths of 355 and 515 nm, respectively. The lower limit of quantitation (LLQ) in blood was established at 5.00 ng/ml for both compounds, with values for within-run precision (WRP) and between-run precision (BRP) of less than 10%. The method is currently being applied to investigate the blood distribution of CPT-11 and SN-38 in cancer patients.

Determination of irinotecan (CPT-11) and its active metabolite SN-38 in human plasma by reversed-phase high-performance liquid chromatography with fluorescence detection

Sensitive high-performance liquid chromatographic assays have been develope to determine the levels of the lactone and lactone plus carboxylate (total) forms of the antitumor agent irinotecan (CPT-11) and its active metabolite SN-38, in human plasma. The related compound camptothecin was used as the internal standard. The selective sample pretreatment for the lactone forms involved a single solvent extraction with acetonitrile- n-butyl chloride (1:4,v/v), whereas the sample clean-up for the total forms was a simple protein precipitation with aqueous perchloric acid-methanol (1:1, v/v), which results in the conversion of the carboxylate to the lactone forms. Chromatography was carried out on a Hypersil ODS column, with detection performed fluorimetrically. The methods have been validated, and stability tests under various conditions have been performed. The lower limits of quantitation are 0.5 and 2.0 ng/ml for the lactone and total forms, respectively. The assays have been used in a single pharmacokinetic experiment in a patient to investigate the applicability of the method in vivo.