Quantification of Acyclovir in Human Plasma by Ultra-High-Performance Liquid Chromatography - Heated Electrospray Ionization - Tandem Mass Spectrometry for Bioequivalence Evaluation

Abstract

Pharmacokinetic studies are essential towards determining bioequivalence and establishing pharmacokinetic profiles for drug moieties requires accurate quantification. We report a rapid, sensitive, and robust method for the determination of acyclovir in human plasma and its validation towards evaluating the bioequivalence of drug formulations. After a simple liquid-liquid extraction from plasma, acyclovir is quantified using ultra-high-performance liquid chromatography -heated electrospray ionization -tandem mass spectrometry (UHPLC-HESI-MS/MS). The assay has a total analysis time is 5 min, a linear range of 1.0 -2000 ng/mL, a lower limit of detection of 0.5 ng/ mL, and a lower limit of quantification of 1.0 ng/mL. Intra-and inter-day precision is no more than 10.3% and intraand inter-day accuracy was within 13% at various concentrations in human plasma. Validation according to FDA guidelines for bioanalysis indicates that the described UHPLC-HESI-MS/MS method provides rigorous quantification of acyclovir in human plasma and representative data demonstrates successful application towards the determination of pharmacokinetic profiles as part of an evaluation of drug formulation bioequivalence.

Effect of Common Excipients on the Oral Drug Absorption of Biopharmaceutics Classification System Class 3 Drugs Cimetidine and Acyclovir

“…Chromatographic separation of acyclovir was achieved using ultra-HPLC-heated electrospray ionization-tandem mass spectrometry (UHPLC-HESI-MS/MS). 23 The UHPLC system was comprised of an Accela degasser, quaternary pump, and an HTC PAL thermostatted autosampler (Thermo Scientific, San Jose, CA). The column was a Waters BEH C18 50 Â 2.1 mm 2 with 1.7 mm particle size.…”

Quantification of acyclovir in dermal interstitial fluid and human serum by ultra‐high‐performance liquid–high‐resolution tandem mass spectrometry for topical bioequivalence evaluation

“…The analyte response at the LLOQ should be at least 5 times the response compared with the blank response (Araujo, ). In this work, the analyte response was 6 times the response of the blank signal compared with the currently most sensitive method that reported an LLOQ of 1 ng/mL for acyclovir in human plasma (Shao, ). If this LLOQ had been applied to the samples in our study, acyclovir would not have been quantifiable in >75% of the samples.…”

“…Existing analytical methods for acyclovir are based on high-performance liquid chromatography (HPLC) with ultraviolet (Bahrami, Mirzaeei, & Kiani, 2005;Hasan, Chander, Ali, Baboota, & Ali, 2011) or fluorescence detection (Dao, Jiao, & Zhong, 2008), but these methods lack both selectivity and sensitivity. More recent methods that are based on HPLC-tandem mass spectrometry (HPLC-MS/MS) in combination with protein precipitation or solid-phase extraction (SPE) show improved sensitivity, which results in lower limits of quantification (LLOQ) (Holkar, Daphal, Yadav, & Rokade, 2012;Kanneti, Rajesh, Aravinda Raj, & Bhatt, 2009;Rigo-Bonnin et al, 2014;Shao, 2012). These methods, however, are not suitable for the determination of acyclovir in dOFM samples because they require high sample volumes for the sample preparation process.…”

Determination of Acyclovir in Human Plasma Samples by HPLC Method with UV Detection: Application to Single-Dose Pharmacokinetic Study

“…A number of assay methods have been reported for determination of acyclovir in biological fluids using capillary electrophoresis [ 2 ] or liquid chromatographic methods with pulsed amperometric detection [ 3 ], tandem mass spectrometry [ 4 ], fluorescence detection [ 5 - 7 ] or ultraviolet detection [ 8 - 15 ]. In the published methods, liquid-liquid extraction with acetonitrile or mixture of isopropyl alcohol and dichloromethane as solvent has been used for sample preparation [ 4 , 8 , 9 ]. The disadvantage of these methods employing liquid-liquid extraction (with grate chemical consumption) of acyclovir from biological fluids is that they involve several steps yielding poor separation from the serum endogenous interferences.…”