Simultaneous determination of four immunosuppressants by means of high speed and robust on-line solid phase extraction–high performance liquid chromatography–tandem mass spectrometry

Koal, Therese; Deters, Michael; Casetta, Bruno; Kaever, Volkhard

doi:10.1016/j.jchromb.2004.02.040

Cited by 117 publications

(79 citation statements)

References 26 publications

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“…Under these simplified chromatographic conditions, CsA and CsD co-eluted with the unspecified, more hydrophilic metabolites of CsA; one single peak was found and integrated in both MRM traces ( Figure 1C, D), both in calibration samples and patient samples. The retention time of this single peak was approximately 2.1 min, which is in agreement with previously described methods for the quantification of CsA employing CsD as the internal standard and the same mass transitions as applied here (2)(3)(4)(5). The quantitative results obtained in this series using limited chromatography differed markedly from those obtained with extended chromatography and from the immunoassay results (Table 1).…”

supporting

confidence: 89%

Pitfall in the high-throughput quantification of whole blood cyclosporin A using liquid chromatography-tandem mass spectrometry

Vogeser

Spohrer²

2005

Clinical Chemistry and Laboratory Medicine (CCLM)

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In a growing number of laboratories the technique of liquid chromatography-tandem mass spectrometry is used for the quantification of cyclosporin A in whole blood, employing cyclosporin D as the internal standard. Cyclosporin A is extensively metabolized in vivo; in liquid chromatography-tandem mass spectrometry respective metabolites can give rise to both parent and product ions that are isobaric with ions commonly used for the detection of cyclosporin A and cyclosporin D, respectively. In this article it is demonstrated that limited chromatography with co-elution of such metabolites together with cyclosporin A and cyclosporin D can lead to incorrect results.Keywords: cyclosporin A; cyclosporin D; liquid chromatography-tandem mass spectrometry; specificity.In tandem-mass spectrometry the detection of target analytes is based on their physical decomposition behavior; the molecular mass-specific selection of an intact precursor ion of an analyte and of the respective product ion results in very high analytical specificity. This high degree of specificity can limit the requirements of sample preparation and chromatography and may enable ''high-throughput'' routine methods. However, the specificity of tandem-mass spectrometry should not be assumed to be absolute and unlimited. Given the complexity of human body fluids as a matrix and the extensive metabolization of certain target analytes, the specificity of liquid chromatography-tandem mass spectrometry (LC-MS/MS) may be a critical issue. This is demonstrated by an observation reported here.With the intention of switching the quantification of whole blood cyclosporin A (CsA) from immunoassay technology to LC-MS/MS, we decided to modify the LC-MS/MS method that we currently use for the quan-

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supporting

confidence: 89%

Pitfall in the high-throughput quantification of whole blood cyclosporin A using liquid chromatography-tandem mass spectrometry

Vogeser

Spohrer²

2005

Clinical Chemistry and Laboratory Medicine (CCLM)

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“…16 Standard working solutions of cyclosporine at concentrations of 100, 200, 500, 1000, 2000, 5000, and 10000 ng mL -1 , and a 10000 ng mL -1 solution of the IS, were prepared by serial dilution of stock solutions. The diluent consisted of a mixture of 0.1% formic acid in water and 0.1% formic acid in acetonitrile (50/50, v/v).…”

Section: Preparation Of Standards and Qc Samplesmentioning

confidence: 99%

“…This procedure was much simpler than that used in previous studies. [9][10][11][12]16,17 and resulted in acceptable extraction recovery and precision for both cyclosporine and the IS.…”

Section: Sample Preparationmentioning

confidence: 99%

“…Many studies have been addressed specificity with regard to measurements of cyclosporine levels. [4][5][6][7][8] Recent studies have analyzed cyclosporine using HPLC-MS/ MS. [9][10][11][12] Also, some reports have been published on the simultaneous analysis of several immunosuppressant drugs (sirolimus, everolimus, and tacrolimus) [13][14][15][16][17] or the simultaneous analysis of cyclosporine metabolites. [18][19][20][21] These studies are time-consuming, and require many steps for sample preparation, including evaporation or reconstitution.…”

Section: Introductionmentioning

confidence: 99%

“…[18][19][20][21] These studies are time-consuming, and require many steps for sample preparation, including evaporation or reconstitution. [9][10][11][12]16,17 UPLC-MS/MS has been used as an alternative to traditional HPLC, and has revolutionized the analysis of biological samples. UPLC provides increased resolution and sensitivity, reduced analysis time, and a lower cost per assay than HPLC.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Quantitative Determination of Cyclosporine in Human Whole Blood by Ultra-Performance Liquid Chromatography with Triple Quadrupole Tandem Mass Spectrometry

et al. 2014

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Cyclosporine is an immunosuppressant drug used in organ transplants or for the treatment of autoimmune diseases. We developed and validated a simple, sensitive, and specific method using UPLC-MS/MS to determine cyclosporine levels in human whole blood. MS/MS detection was performed in the positive electrospray ionization mode with multiple reaction monitoring. Cyclosporine was extracted from whole-blood samples using ascomycin as an internal standard. The mass transitions m/z 1203.49 → 1185.53 and m/z 814.71 → 796.67 were used to assay the analyte and IS. This method was validated with respect to linearity, specificity, accuracy, precision, recovery, and stability. The method exhibited a linear response from 10 to 1000 ng mL -1 with correlation coefficient values >0.99. The precision and the accuracy values were within 15%, except at the lower limit of quatification (LLOQ). Cyclosporine was stable in whole blood with no evidence of degradation. This method was successfully applied to a pharmacokinetic study of cyclosporine in healthy volunteers following oral administration.Keywords Cyclosporine, UPLC-MS/MS, human whole blood

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Liquid chromatographic method for the determination of sirolimus in blood using electrochemical detection

Mochizuki

Suka

Matsumoto

et al. 2008

Biomedical Chromatography

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Therapeutic drug monitoring of sirolimus (rapamycin) is important for immunosuppressive therapy in solid organ transplantation. We have developed a simple and reliable method for determining blood concentrations of sirolimus using reversed-phase HPLC with electrochemical detection (ECD). The E(2) potential was set at +900 mV. The potential of guard cell was set at +950 mV and that of the E(1) cell at +400 mV. The method was linear for a concentration range of 1-50 ng/mL when 0.5 mL blood was used. The correlation coefficients of all standard curves were greater than or equal to 0.999. The limit of detection was 0.5 ng/mL. The inter-assay precision ranged from 3.22 to 7.48%, and the coefficient of variation (CV) for a quality control sample at 10 ng/mL was 7.48% with a bias of 8.4% from the target value. The intra-assay precision ranged from 0.72 to 3.71%, and the CV for a quality control sample at 10 ng/mL was 0.72% with a bias of 6.8% from the target value. In a solid organ transplant recipient, trough concentrations of sirolimus were well within the analytic range of the HPLC/ECD procedure. The method described here is suitable for management of sirolimus therapy in solid organ transplantation.

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Simultaneous determination of four immunosuppressants by means of high speed and robust on-line solid phase extraction–high performance liquid chromatography–tandem mass spectrometry

Cited by 117 publications

References 26 publications

Pitfall in the high-throughput quantification of whole blood cyclosporin A using liquid chromatography-tandem mass spectrometry

Pitfall in the high-throughput quantification of whole blood cyclosporin A using liquid chromatography-tandem mass spectrometry

Quantitative Determination of Cyclosporine in Human Whole Blood by Ultra-Performance Liquid Chromatography with Triple Quadrupole Tandem Mass Spectrometry

Liquid chromatographic method for the determination of sirolimus in blood using electrochemical detection

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