Advantages of using tetrahydrofuran–water as mobile phases in the quantitation of cyclosporin A in monkey and rat plasma by liquid chromatography–tandem mass spectrometry

The aim of the study was to develop and validate a simple and rapid method for the determination of cyclosporine A (CsA) in ocular rabbit tissues using reversed-phase ultra-high-performance liquid chromatography (UHPLC) with UV detection. Previous publications on chromatographic methods of CsA determination in ocular tissues involved only reversed-phase HPLC separation, usually in combination with such detection techniques as radio-immunoassay and mass spectrometry. The application of the UHPLC technique allowed us to significantly decrease the analysis time. Cyclosporine D (CsD) was applied as the internal standard. Satisfactory separation was achieved on an XB-C18 Kinetex column at 60°C with the use of gradient elution mode. The retention times of CsA and CsD were found to be 4.5 and 5.1 min, respectively. The developed assay is specific, sensitive (limit of detection = 6 ng/mL and limit of quantitation = 18 ng/mL) and linear within the analyte concentration range of 0.018-5 µg/mL, with a correlation coefficient of 0.999. High sensitivity, low injection volume (10 μL), short time of analysis (6.5 min) and simplicity make this method useful for the fast analysis of CsA in rabbit ocular tissues and fluids: lacrimal fluid, aqueous humor, cornea, conjunctiva and eye globe.

Section: Resultsmentioning

confidence: 99%

Development and validation of UHPLC method for the determination of cyclosporine A in biological samples

Szerkus

Wolska

Struck-Lewicka

et al. 2014

“…Very few LC-MS methods have been reported for CyA in rat plasma. Li et al (2007) described a CyA LC-MS assay for use in monkey and rat plasma, although cyclosporine C was used as IS. Similarly Shibata et al (2004) reported a method using cyclosporine D as IS; as mentioned above, neither of these two IS is available commercially.…”

Section: Discussionmentioning

confidence: 99%

“…For research purposes, the specific measurement of CyA is possible using one of a number of valuable highperformance liquid chromatographic (HPLC) methods (Burckart et al, 1990;Chimalakonda et al, 2002;McBride et al, 1989). More specific and sensitive liquid chromatography-mass spectrometry (LC-MS) methods have been reported for the measurement of CyA in various matrices, although many incorporate solid phase extraction (Zaater et al, 2005) or commercially unavailable internal standards (IS) such as cyclosporine C and D (Li et al, 2007;Shibata et al, 2004;Simpson et al, 1998), which can pose challenges to their widespread use.…”

Section: Introductionmentioning

confidence: 99%

An analytical method for cyclosporine using liquid chromatography–mass spectrometry

Kanduru

Somayaji

Lavasanifar

et al. 2009

A liquid chromatographic mass spectrometric (LC-MS) assay has been developed for cyclosporine A (CyA) in rat plasma using amiodarone as internal standard (IS). Rat plasma (100 microL) containing drug and IS were extracted using liquid-liquid extraction with 4 mL of 95:5 ether:methanol. After evaporation of the organic layer the residue was reconstituted with 500 microL of water. Then the aqueous layer was transferred to LC-MS sample vials. A 10 microL volume was injected. The analysis was performed on a C(8) column 3.5 microm (2.1 x 50 mm) heated to 60 degrees C with a mobile phase consisting of acetonitrile:methanol:0.2% NH(4)OH (60:20:20) at an isocratic flow-rate of 0.2 mL/min. The ions used for quantitation of CyA and IS were m/z 1202.8 and 645.9, with retention times of 3.35 and 4.72 min, respectively. Linear relationships (r(2) > 0.99) were achieved between plasma or blood concentration and peak height ratios (drug:IS) over the concentration range 50-5000 ng/mL. The CV% and mean error were <19%. Based on validation data, the lower limit of quantification for the assay was 50 ng/mL. The reported assay method displayed high measures of linearity, sensitivity, reliability and precision, allowing its applicability in pharmacokinetic studies in rat.

Development and validation of an LC‐ESI‐MS/MS approach to determine a highly hydrophobic drug, norcantharidin palmitate, and apply to a preliminary pharmacokinetic study in rats

Liu

Tao

Zheng

et al. 2017

In order to investigate the pharmacokinetics of norcantharidin palmitate (NCTD-PAL) in rats, we developed and validated an LC-ESI-MS/MS method. The NCTD-PAL and internal standard (triamcinoloneacetonide palmitate, TAP) were separated on a Phenomenex Kinetex®XB C column, and the mobile phase was composed of tetrahydrofuran (THF)-acetonitrile (20/80, v/v) and an aqueous phase containing 0.2% ammonium hydroxide at a flow rate of 0.3 mL/min. The ESI interface operated in positive mode was used to acquire the mass spectrometric data, and the transition ions were m/z 635.50 → 168.95 and 673.65 → 397.13 for NCTD-PAL and IS, respectively. The method had a linear range of 10-2000 ng/mL with a correlation coefficient of >0.99. The accuracy (RE, %) was within ±10.1%, and the intra- and inter-day precisions (RSD, %) were 10.9 and 13.8%, respectively. The extraction recovery of NCTD-PAL at different concentrations ranged from 89.3 to 102.0%. The validated approach was efficaciously applied to a pharmacokinetic study of NCTD-PAL in rats via intravenous injection. Based on these results obtained, this method is practical and suitable for a wide range of applications.