Measurement of citrate in urine using liquid chromatography tandem mass spectrometry: comparison with an enzymatic method

2005 ◽  
Vol 42 (5) ◽  
pp. 357-363 ◽  
Author(s):  
BG Keevil ◽  
L Owen ◽  
S Thornton ◽  
J Kavanagh
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Ammonium Acetate ◽  
Enzymatic Assay ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Urinary Citrate

Background: Measurement of urine citrate is used to assess the risk of further urinary stone formation and to assess the benefit of treatment in affected individuals. We wanted to develop a simple and rapid liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the analysis of urinary citrate and to compare it with our current enzymatic assay. Methods: For the LC-MS/MS assay, samples were prepared in a deep-well block by adding 10 µL of urine and 20 µL of internal standard to 400 µL of water. After mixing, 3 µL of the diluted sample was injected into the LC-MS/MS system. An LC system was used to isocratically elute a C18 column (50 x 2.1 mm) with 0.4 mL/min water containing 2 mmol/L ammonium acetate and 0.1% (v/v) formic acid. A step gradient of 100% methanol containing 2 mmol/L ammonium acetate and 0.1% (v/v) formic acid was used to wash the column. The retention times were 1.4 min for citrate and 1.4 min for d4-citrate. Cycle time was 4.0 min, injection to injection. The analytes were monitored using a tandem mass spectrometer operated in multiple reaction monitoring mode using the following transitions, citrate m/ z 191.0> 111.0 and d4-citrate m/ z 195.0> 113.0. Results: Within and between-batch coefficients of variation were <3% over the range 480-3800 µmol/L. The lower limit of quantification was 24.0 µmol/L. Regression analysis showed LC-MS/MS = 0.8781 (enzymatic assay) + 102.5, r = 0.964, n = 73. Conclusions: We have developed a simple LC-MS/MS method for urinary citrate measurement that shows acceptable performance.

scholarly journals Simultaneous Determination of Cortisol and Cortisone from Human Serum by Liquid Chromatography-Tandem Mass Spectrometry

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Sanghoo Lee ◽  
Hwan-Sub Lim ◽  
Hye-Jin Shin ◽  
Seol-A Kim ◽  
Jimyeong Park ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Ammonium Acetate ◽  
Cortisol Concentration ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Cortisol Ratio

A fast, sensitive, and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was validated and then the levels of cortisol and cortisone from sera of healthy adults were determined by the LC-MS/MS method. One hundredμL of serum sample was directly extracted by adding 2 mL ethyl acetate, followed by chromatographic separation on a C18 column with a mobile phase consisting of 5 mM ammonium acetate and methanol (25 : 75, v/v). The precision, accuracy, and average recovery of the method were 1.5–5.3%, 95.4–102.5%, and 96.4% for cortisol, and 1.9–6.0%, 89.2–98.8%, and 79.9% for cortisone, respectively. The method was linear from 1.0 to 500.0 ng/mLr2=0.999for cortisol and 2.5 to 100.0 ng/mLr2=0.998for cortisone. The limits of detection (LOD) and quantification (LOQ) were 0.2 and 1.0 ng/mL for cortisol, and 1.0 and 2.5 ng/mL for cortisone, respectively. The average cortisol concentration (133.9±63.7 ng/mL) of samples collected between 9:00 and 11:00 a.m. was higher approximately 4.4 times than that of cortisone (30.5±10.7 ng/mL)P<0.0001. The average cortisone/cortisol ratio was 0.225. Therefore, the LC-MS/MS method may be useful for the diagnosis of some adrenal diseases and the assessment of 11β-hydroxysteroid dehydrogenase (11β-HSD) activity in clinical laboratories.

scholarly journals Liquid chromatography – tandem mass spectrometry method for the determination of ten tetracycline residues in muscle samples

2015 ◽  
Vol 59 (3) ◽  
pp. 345-352 ◽  
Author(s):  
Anna Gajda ◽  
Andrzej Posyniak
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Solid Phase ◽  
Acid Extraction ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

Abstract A liquid chromatography – tandem mass spectrometry (LC-MS/MS) method for the determination of oxytetracycline (OTC), 4-epi oxytetracycline (4-epi OTC), tetracycline (TC), 4-epi tetracycline (4-epi TC), chlortetracycline (CTC), 4-epi chlortetracycline (4-epi CTC), doxycycline (DC), minocycline (MINO), methacycline (META) and rolitetracycline (ROLI) residues in muscles was developed. The procedure consisted of an oxalic acid extraction followed by protein removal with trichloroacetic acid. Further solid phase clean-up on polymeric (Strata X) reversed phase columns was performed to obtain an extract suitable for LC-MS/MS analysis. The tetracyclines were separated on a C 18 analytical column with mobile phase consisting of 0.01% formic acid in acetonitrile and 0.01% formic acid in water in gradient mode. The method was validated according to the Commission Decision 2002/657/EC. The recoveries of all target compounds were 91.8% – 103.6%. The decision limits were from 109.0 to 119.8 μg/kg and detection capability varied within the range of 122.2 to 137.6 μg/kg, depending on the analyte.

scholarly journals ANALYSIS OF 6-MERCAPTOPURINE AND 6-METHYLMERCAPTOPURINE IN DRIED BLOOD SPOTS USING LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY AND ITS APPLICATION IN CHILDHOOD ACUTE LYMPHOBLASTIC LEUKEMIA PATIENTS

2017 ◽  
Vol 10 (9) ◽  
pp. 120 ◽  
Author(s):  
Supandi Supandi ◽  
Yahdiana Harahap ◽  
Harmita Harmita ◽  
Rizka Andalusia ◽  
Marlina Ika
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Maintenance Phase ◽  
Dried Blood Spots ◽  
Tandem Mass ◽  
Blood Spots ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

  Objective: To analyze and validate 6-mercaptopurine (6-MP) and 6-methylmercaptopurine (6-MMP) in dried blood spots (DBS) using liquid chromatography-tandem mass spectrometry (LC/MS-MS).Methods: Bio-sampling dried blood spot with DBS-CAMAG® paper diameter of 8 mm and extracted with acetonitrile-methanol (1:3) containing internal standard 5-fluorouracil (5-FU). Separation was performed with C18 column Acquity® 1.7 μm (2.1 mm × 100 mm), with a mobile phase mixture of 0.1% formic acid in water 0.1% formic acid in acetonitrile with gradient elution and flow rate 0.2 ml/min. Mass detection was Waters Xevo TQD with positive electrospray ionization (ESI) for 6-MP, 6-MMP and negative ESI for 5-FU in multiple reaction monitoring modes. The ions of 6-MP was detected at m/z 153.09->119.09, 6-MMP at m/z 167.17->126.03, and 5-FU at m/z 129.15->42.05.Results: This method fulfill the requirements of selectivity, linearity, lower limit of quantification, accuracy, precision, carry-over, matrix effects, and stability which refers to the european medicines agency (EMEA) guidelines. The linearity of 0.99 was 26-1000 ng/mL for 6-MP and 6-MMP, respectively. The validated method was applied to two childhood ALL maintenance phase. Retrieved 6-MP levels of 10.37 pmol/8×108 erythrocytes, respectively. The levels of 6-MMP gained 16.59 pmol/8×108 erythrocytes, respectively.Conclusion: The developed LC/MS-MS method is valid to analysis 6-MP and 6-MMP in DBS simultaneous in vitro according to EMEA guidelines. The method was successfully applied to authentic capillary blood samples from two childhood patients with ALL in the maintenance phase.

scholarly journals Simultaneous Analysis of 6-Mercaptopurine, 6-Methylmercaptopurine, and 6-Thioguanosine-5’-monophosphate in Dried Blood Spot Using Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

2018 ◽  
Vol 18 (3) ◽  
pp. 544 ◽  
Author(s):  
Supandi Supandi ◽  
Yahdiana Harahap ◽  
Harmita Harmita ◽  
Rizka Andalusia
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Ultra Performance Liquid Chromatography ◽  
Dried Blood Spot ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Blood Spot

6-Mercaptopurine is a chemotherapeutic agent of the antimetabolite class. This study aims to analyze simultaneous validation of 6-mercaptopurine (6-MP), 6-methylmercaptopurine (6-MMP), and 6-thioguanosine-5’-monophosphate (6-TGMP) in dried blood spot (DBS) using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). An accurate volume of 60 μL blood was spotted onto DBS-CAMAG paper and then extracted using methanol 90% (v/v) containing an internal standard of 5-fluorouracil (5-FU). Separation was performed using a Waters Acquity UPLC BEH AMIDA column 1.7 μm (2.1 x 100 mm) with a mobile phase mixture of 0.2% (v/v) formic acid in water−0.1% (v/v) formic acid in acetonitrile-methanol with gradient elution and flow rate of 0.2 mL/min. Mass detection was done using Waters Xevo TQD with positive electrospray ionization (ESI) for 6-MP, 6-MMP, 6-TGMP and negative ESI for 5-FU, in multiple reaction monitoring mode. Detection rates of 6-MP, 6-MMP, 6-TGMP and 5-FU were m/z 153.09 > 119.09; 167.17 > 126.03; 380.16 > 168.00); 129.09 > 42.05, respectively. This method is linear across the range 25.5–1020 ng/mL for 6-MP, 6-MMP and 6-TGMP. This method is valid for the in vitro simultaneous analysis of 6-MP, 6-MMP and 6-TGMP in DBS, based on European Medicine Agency guidelines.

scholarly journals Analysis of Methylmalonic Acid in Plasma by Liquid Chromatography–Tandem Mass Spectrometry

2006 ◽  
Vol 52 (4) ◽  
pp. 754-757 ◽  
Author(s):  
Anne Schmedes ◽  
Ivan Brandslund
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Methylmalonic Acid ◽  
Cobalamin Deficiency ◽  
Tandem Mass ◽  
Simple Method ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

Abstract Background: Methylmalonic acid (MMA) is a biochemical marker for cobalamin deficiency, particularly in cases where the cobalamin concentration is moderately decreased or in the low-normal range. Liquid chromatography–tandem mass spectrometry (LC-MS/MS) with electrospray ionization is a rapid, robust method that has been used in MMA analysis. We developed a simple method combining solid-phase extraction (SPE) and derivatization to prepare serum or plasma for LC-MS/MS analysis of MMA. Methods: Deuterated internal standard d3-MMA was added to serum or plasma before SPE on strong anion-exchange (SAX) columns. After elution with HCl–butanol (10:90 by volume) and addition of 1 g/L formic acid, the samples were simultaneously derivatized and evaporated by heating to 70 °C for 15 min followed by 54 °C overnight in uncapped vials. Acetonitrile and 1 g/L formic acid were added to the samples before injection into the LC-MS/MS system. MMA and d3-MMA were quantified in the multiple-reaction monitoring mode. Calibrators were prepared in serum by the standard addition method. Results: The MMA assay was linear up to 200 μmol/L. Interassay CVs were 6.7%, 5.0%, and 5.0% for mean concentrations of 0.15, 0.36, and 0.65 μmol/L, respectively. Conclusions: Our simplified sample preparation and derivatization method is suitable for use in MMA analyses. MMA elutes with the derivatization reagent, and derivatization and evaporation are performed simply by leaving the uncapped vials in a heating block overnight. The method shows good linearity and precision.

scholarly journals Development and Validation of High-Throughput Bioanalytical Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) Method for the Quantification of Newly Synthesized Antitumor Carbonic Anhydrase Inhibitors in Human Plasma

Molecules ◽  
2020 ◽  
Vol 25 (23) ◽  
pp. 5753
Author(s):  
Ahmed M. Abdel-Megied ◽  
Wagdy M. Eldehna ◽  
Mohamed A. Abdelrahman ◽  
Fawzy A. Elbarbry
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Human Plasma ◽  
Linear Regression Method ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

In the present study, a sensitive and fully validated bioanalytical high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the quantitative determination of three newly synthesized carbonic anhydrases inhibitors (CAIs) with potential antitumor activity in human plasma. The analytes and the internal standard (IS) were extracted using 1.5 mL acetonitrile from only 450 µL aliquots of human plasma to achieve the desired protein precipitation. Chromatographic separations were achieved on Phenomenex Kinetex® C18 column (100 × 4.6 mm, 2.6 µm) using a binary gradient elution mode with a run time of less than 6 min. The mobile phase consisted of solvent (A): 0.1% formic acid in 50% methanol and solvent B: 0.1% formic acid in acetonitrile (30:70, v/v), pumped at a flow rate of 0.8 mL/min. Detection was employed using triple quadrupole tandem mass spectrometer (API 3500) equipped with an electrospray ionization (ESI) source in the positive ion mode. Multiple reaction monitoring (MRM) mode was selected for quantitation through monitoring the precursor-to-parent ion transition at m/z 291.9 → 173.0, m/z 396.9 → 225.1, m/z 388.9 → 217.0, and m/z 146.9 → 91.0 for AW-9a, WES-1, WES-2, and Coumarin (IS), respectively. Linearity was computed using the weighted least-squares linear regression method (1/x2) over a concentration range of 1–1000, 2.5–800, and 5–500 ng/mL for AW-9a, WES-1, and WES-2; respectively. The bioanalytical LC-MS/MS method was fully validated as per U.S. Food and Drug Administration (FDA) guidelines with all respect to linearity, accuracy, precision, carry-over, selectivity, dilution integrity, and stability. The proposed LC-MS/MS method was applied successfully for the determination of all investigated drugs in spiked human plasma with no significant matrix effect, which is a crucial cornerstone in further therapeutic drug monitoring of newly developed therapeutic agents.

scholarly journals FACTORS AFFECTING LIQUID CHROMATOGRAPHY TANDEM-MASS SPECTROMETRY ANALYSIS OF 6-MERCAPTOPURINE AND 6-THIOGUANINE IN DRIED BLOOD SPOTS

2018 ◽  
Vol 10 (1) ◽  
pp. 200
Author(s):  
Yahdiana Harahap ◽  
Dimas Agus Putera Hardijanto ◽  
Delly Ramadon
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Internal Standard ◽  
Standard Addition ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Blood Spot

Objective: This study aimed to determine the effects of the method of internal standard addition, spotting volume, paper type, and sample storagetemperature on 6-mercaptopurine, and 6-thioguanine on liquid chromatography tandem-mass spectrometry (LC-MS/MS) bioanalysis methods usingdried blood spot (DBS).Methods: Blood samples were spotted on CAMAG DBS paper and a Perkin Elmer 226 sample collection device (paper) and extracted into methanolcontaining 5-fluorouracil as an internal standard. The separation was performed on a water acquity ultra high-performance LC BEH Amide 1.7 μm(2.1 mm×100 mm) column with a mobile phase of 0.2% formic acid in water - 0.1% formic acid in acetonitrile methanol with gradient elution at aflow rate of 0.2 mL/min.Results: The step at which the internal standard was added (blood, spot on DBS card, or extraction solution) affected the chromatogram. Differencesin paper types and blood volumes significantly affected (p<0.05) the percent coefficient of variation, whereas differences in blood hematocritsignificantly affected the peak area ratio.Conclusion: The method of internal standard addition affected the chromatograms in this study. The best chromatogram was observed whenthe Internal Standard was added to the extracting solution. The card type also affected the analysis, so it is recommended to use the same card duringsample analysis.

scholarly journals ANALYTICAL METHOD VALIDATION OF DOXORUBICIN AND DOXORUBICINOL IN VOLUMETRIC ABSORPTIVE MICROSAMPLING BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY

2021 ◽  
Vol 56 (5) ◽  
pp. 424-432
Author(s):  
Yahdiana Harahap ◽  
Maria Juanita ◽  
Baitha Palanggatan Maggadani
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Sample Preparation ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Analytical Method ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Volumetric Absorptive Microsampling

Doxorubicin is a broad-spectrum anthracycline antibiotic which has antineoplastic activity. Doxorubicin can cause cardiotoxic effects due to the formation of doxorubicinol as its main metabolite. One of the latest bio sampling methods, Volumetric Absorptive Microsampling (VAMS), has various advantages which include blood sampling by finger-prick, hematocrit does not influence it, and it can be stored at room temperature. This study aims to determine the optimum analysis conditions and sample preparation method in VAMS with daunorubicin as an internal standard, and to develop sensitive, specific measurements as well as validate the analytical method using Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS). Analytical quantification analysis using mass spectrometry with a mass analyzer type is a triple quadrupole with a positive type of electrospray ionization (ESI). The separation was carried out with the Acquity® UPLC BEH C18 column (2.1 x 100 mm; 1.7 μm), with a flow rate of 0.2 mL/min, and gradient elution using 0.1% formic acid in water and 0.1% formic acid in acetonitrile for 7 minutes. Multiple reaction monitoring (MRM) values were set at m/z 544.22 > 396.9 for doxorubicin; m/z 546.22 > 398.9 for doxorubicinol; and m/z 528.5 > 362.95 for daunorubicin. Sample preparation used protein precipitation with methanol as the extracting solution, the drying time of the VAMS was 2 hours, and the sonication time was 30 minutes. LLOQ values obtained were 8 ng/mL for doxorubicin and 3 ng/mL for doxorubicinol with the linearity of 0.9904 for doxorubicin and 0.9902 for doxorubicinol.

scholarly journals Simple and Sensitive Analysis of Blonanserin and Blonanserin C in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry and Its Application

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Yunliang Zheng ◽  
Xingjiang Hu ◽  
Jian Liu ◽  
Guolan Wu ◽  
Huili Zhou ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Human Plasma ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

A highly sensitive, simple, and rapid liquid chromatography tandem mass spectrometry method to simultaneously determine blonanserin and blonanserin C in human plasma with AD-5332 as internal standard (IS) was established. A simple direct protein precipitation method was used for the sample pretreatment, and chromatographic separation was performed on a Waters XBridge C8(4.6×150 mm, 3.5 μm) column. The mobile phase consists of a mixture of 10 mM ammonium formate and 0.1% formic acid in water (A) and 0.1% formic acid in methanol (B). To quantify blonanserin, blonanserin C, and IS, multiple reaction monitoring (MRM) was performed in positive ESI mode. The calibration curve was linear in the concentration range of 0.012–5.78 ng·mL−1for blonanserin and 0.023–11.57 ng·mL−1for blonanserin C (r2>0.9990). The intra- and interday precision of three quality control (QC) levels in plasma were less than 7.5%. Finally, the current simple, sensitive, and accurate LC-MS/MS method was successfully applied to investigate the pharmacokinetics of blonanserin and blonanserin C in healthy Chinese volunteers.

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