Recent development in application of high performance liquid chromatography-tandem mass spectrometry in therapeutic drug monitoring of immunosuppressants

Many routine hospital and clinical research assays have relied upon immunoassay procedures to achieve sensitive measurements of a range of important analytes. Some of the methods have been developed in-house but increasingly commercial kits and automated analysers have become commonplace. The accuracies of these methods are under question in health care. Mass spectrometry (MS) is potentially a more accurate technique with the ability to demonstrate specificity. An introduction of the basic analytical aspects of liquid chromatography (LC) -MS/MS leads on to the validation of the method before general use. LC coupled with MS and tandem mass spectrometry (MS n ) is being adopted in a number of hospital laboratories for the quantitative analysis of a number of analytes from physiological matrices, but standards for development and validation of such assays are not easily available. Most assays can be regarded as in-house methods and herein may lay the failure so far for mass spectrometric methods to improve quality of results between laboratories for an analyte using the same technology. Manufacturers are taking on board the experience of clinical laboratories with kits containing all or most of the disposable items and reagents. A number of documents and guidelines have been consulted. These documents are expensive to purchase, are often very long and not easy to read. This review highlights the specific requirements for introduction of a tandem mass spectrometric test for small molecules into a routine hospital laboratory. A number of experiments need to be planned and executed in order to describe a new quantitative method in terms of selectivity, accuracy, imprecision, sensitivity and stability. The introduction of a quantitative method based on tandem MS requires careful validation. This review has distilled out important points from a number of key documents in order to provide a working validation guideline for clinical laboratories. In a supplementary file a working document for assembling the assay validation is proposed.

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“…ascomycin in acetonitrile). 8,62 Methanol used in HPLC eluting solutions can cause ion suppression. 63 …”

Section: Ruggednessmentioning

confidence: 99%

Development and validation of a quantitative assay based on tandem mass spectrometry

Honour

2011

Ann Clin Biochem

113

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“…There are also recent development on application of new technologies in immunosuppressant TDM including sample preparation and ionization (Yang & Wang, 2008). Dried blood spot sampling is an alternative to venous blood sampling for immunosuppressant TDM due to the convenient sample collection from fingerprick and small blood volume collected.…”

Section: Lc-ms/ms In Therapeutic Drug Monitoring For Immunosuppressantsmentioning

confidence: 99%

Principles and Applications of LC-MS/MS for the Quantitative Bioanalysis of Analytes in Various Biological Samples

Kang¹

2012

Tandem Mass Spectrometry - Applications and Principles

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“…Although immunoassays offer rapid analysis and easy handling, there are many concerns about these methods because of the cross-reactivity of the antibodies used with inactive CyA metabolites that may result in overestimation of the drug values [12][13][14]. For this reason, chromatography-based methods such as high performance liquid chromatography with ultraviolent detection (HPLC-UV), high performance liquid chromatography coupled to mass spectrometry detection (HPLC-MS) and ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), are considered the best options for the quantification of CyA in biological matrices [15][16][17]. HPLC-UV has some limitations related to specificity and sensibility.…”

Section: Introductionmentioning

confidence: 99%

“…Consequently, laborious and time-consuming extraction procedures and chromatographical separations are required to improve the limit of drug quantification. During the past decade mass detection has attracted more interest since quantification is based on the relationship of mass/electric charge of the molecule and thus all compounds with different mass to the target do not interfere in the measurement [16]. This leads to both reduced sample preparation and chromatographical separation efforts in complex matrices.…”

Section: Introductionmentioning

confidence: 99%

Ultra high performance liquid chromatography–tandem mass spectrometry method for cyclosporine a quantification in biological samples and lipid nanosystems

Guada

Imbuluzqueta

Mendoza

et al. 2013

Journal of Chromatography B

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Cyclosporine A (CyA) is an immunosuppressant cyclic undecapeptide used for the prevention of organ transplant rejection and in the treatment of several autoimmune disorders. An ultra high performance liquid chromatography-tandem mass spectrometry method (UHPLC-MS/MS) to quantify CyA in lipid nanosystems and mouse biological matrices (whole blood, kidneys, lungs, spleen, liver, heart, brain, stomach and intestine) was developed and fully validated.Chromatographic separation was performed on an Acquity UPLC ® BEH C18 column with a gradient elution consisting of methanol and 2 mM ammonium acetate aqueous solution containing 0.1% formic acid at a flow rate of 0.6 mL/min. Amiodarone was used as internal standard (IS). Retention times of IS and CyA were 0.69 min and 1.09 min, respectively. Mass spectrometer operated in electrospray ionization positive mode (ESI+) and multiple reaction monitoring (MRM) transitions were detected, m/z 1220.69→1203.7 for CyA and m/z 646→58 for IS. The extraction method from biological samples consisted of a simple protein precipitation with 10% trichloroacetic acid aqueous solution and acetonitrile and 5 µL of supernatant were directly injected into the UHPLC-MS/MS system. Linearity was observed between 0.001 µg/mL-2.5 µg/mL (r ≥ 0.99) in all matrices. The precision expressed in coefficient of variation (CV) was below 11.44% and accuracy in bias ranged from −12.78% to 7.99% including methanol and biological matrices. Recovery in all cases was above 70.54% and some matrix effect was observed. CyA was found to be stable in post-extraction whole blood and liver homogenate samples exposed for 6 h at room temperature and 72 h at 4 o C. The present method was successfully applied for quality control of lipid nanocarriers as well as in vivo studies in BALB/c mice.

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Recent development in application of high performance liquid chromatography-tandem mass spectrometry in therapeutic drug monitoring of immunosuppressants

Cited by 69 publications

References 39 publications

Development and validation of a quantitative assay based on tandem mass spectrometry

Development and validation of a quantitative assay based on tandem mass spectrometry

Principles and Applications of LC-MS/MS for the Quantitative Bioanalysis of Analytes in Various Biological Samples

Ultra high performance liquid chromatography–tandem mass spectrometry method for cyclosporine a quantification in biological samples and lipid nanosystems

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