A Conventional LC-MS Method Developed for the Determination of Plasma Raltegravir Concentrations

Abstract

Raltegravir belongs to a new class of antiretrovirals acting for a human immunodeficiency virus (HIV)-1 integrase inhibition. Clinical trials of this drug have demonstrated potent antiviral activity in both therapy naïve and experienced patients. Thus, raltegravir has become an important component of combination treatment regimens used to treat patients with multidrug-resistant HIV-1. The quantification of raltegravir in human plasma is important to support clinical studies and determine pharmacokinetic parameters of raltegravir in HIV-1 infected patients. Recently, the LC-MS/MS superfine system was developed to determine plasma concentration of raltegravir; however, the system needs to be delicately set and the equipment is very expensive. Therefore, we developed a conventional LC-MS method to overcome these difficulties. Subsequently the method was validated by estimating the precision and accuracy for inter-and intraday analysis in the concentration range of 0.010-7.680 m mg/ml. The calibration curve was linear in this range. Average accuracy ranged from 97.2 to 103.4%. Relative standard deviations of both inter-and intraday assays were less than 10.4%. Recovery of raltegravir was more than 80.6%. This novel LC-MS method is accurate and precise enough to determine raltegravir levels in human plasma samples.

Short Communication: Lack of Correlation Between UGT1A1*6, *28 Genotypes, and Plasma Raltegravir Concentrations in Japanese HIV Type 1-Infected Patients

“…Plasma raltegravir concentrations were determined using our previously reported LC-MS equipment and methodology. 10 For the present study, plasma raltegravir concentrations were determined from an average of three separate draws for each patient (range: 2-6 times/individual).…”

Quantification of raltegravir (MK0518) in human plasma by high-performance liquid chromatography with photodiode array detection

“…The LOQ found with our method (10 ng/mL) is higher than the LOQ of other quantification methods known for their greater sensitivity, such as LCMSMS [4,7,8,10,18], but greater than other UV detection quantification methods [12][13][14]. Limit of quantification of the 3 quantification methods using HPLC with UV detection were: 20 ng/mL with 100 L plasma sample and a solid/liquid extraction step for Rezk et al [12]; 23.4 ng/mL with a 500 L plasma sample and a solid/liquid extraction step for D'avolio et al [13], and 19.5 ng/mL with a 600 L plasma sample and a solid/liquid extraction step for Notari et al [14].…”

“…To date nine HPLC analytical methods for quantification of raltegravir have been published. Five of these use liquid chromatography-mass spectrometry [4,[7][8][9][10], one uses fluorescence detection [11], and three others use ultraviolet (UV) detection after a liquid-solid phase [12][13][14]. Here we describe the validation of a simple, sensitive and cost effective HPLC method using photodiode array (PDA) detection and liquid-liquid extraction for quantification of raltegravir.…”

Validation of an UPLC-MS/MS Method for Quantitative Analysis of Raltegravir in Human Plasma Samples

“…[28][29][30][31][32] However, the quality of separation obtained with these columns is affected by higher operating pressures, which require that the system have the components designed to minimize instrumental problems. Many groups use HPLC columns containing particles less than 3.5 mm to achieve faster analyses and increased resolution.…”