Detection of anti-diabetics in equine plasma and urine by liquid chromatography-tandem mass spectrometry.
Abstract: Anti-diabetics such as sulfonylurea and thiazolidinedione derivatives are hypoglycemic drugs used for the treatment of diabetes. However, they can also be used as a stopper in horseracing. This paper describes a convenient method for the separation and simultaneous detection of 10 anti-diabetic drugs (namely glipizide, glibenclamide, glimepiride, gliclazide, tolazamide, tolbutamide, nateglinide, repaglinide, rosiglitazone and pioglitazone) in equine plasma and urine by LC-MS-MS. Methods: The anti-diabetics were isolated from equine plasma and urine by liquid-liquid extraction with 1,2-dichloroethane at acidic pH, and analysed by LC-MS-MS in the positive electrospray ionisation mode. Separation of 10 anti-diabetic drugs was achieved with a reversed phase C8 column using a mixture of aqueous ammonium formate (pH 3.0, 10 mM) and methanol as the mobile phase. Results: Detection and confirmation of the 10 anti-diabetic drugs at 10 ng/mL each in equine plasma and equine urine were achieved by full-scan MS-MS. All of these drugs were detected consistently at this concentration in spiked samples of different plasma and urine (n = 15 each). No significant matrix interferences were observed at the expected retention times of the targeted ions in blank urine samples (n = 30). This method has been used successfully in the analysis of drug-administration samples as well as official racing samples. Conclusions: An LC-MS-MS method has been developed for the simultaneous detection of 10 anti-diabetics in equine plasma and urine. This method can be used to detect the abuse of anti-diabetic drugs in racehorses.
Publication Date: 2004-10-02 PubMed ID: 15458724DOI: 10.1016/j.jchromb.2004.03.070Google Scholar: Lookup
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- Journal Article
Summary
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The research paper presents a successful method for the simultaneous detection of ten types of anti-diabetic drugs in horse plasma and urine, which could be used to prevent unfair practices in horse racing.
Introduction and Objective
- The primary concern of this research paper is the misuse of anti-diabetic drugs, like sulfonylurea and thiazolidinedione derivatives in horse racing. These drugs are typically used for treating diabetes, but in the context of horse racing, they are misused to gain a performance advantage.
- The paper proposes a method that uses Liquid Chromatography Tandem Mass Spectrometry (LC-MS-MS), an effective analytical tool to detect the presence of these ten anti-diabetic drugs in equine plasma and urine.
Methodology
- First, the anti-diabetic drugs were isolated from the horse plasma and urine using a process called liquid-liquid extraction with a chemical compound named 1,2-dichloroethane at acidic pH.
- Then, these samples were analysed with LC-MS-MS using the positive electrospray ionisation mode for better separation and detection of the substances.
- The researchers used a reversed-phase C8 column and a mix of aqueous ammonium formate (pH 3.0, 10 mM) and methanol as the mobile phase to achieve a clear separation of the ten anti-diabetic drugs.
Results
- Through full-scan MS-MS, detection of the ten mentioned anti-diabetic drugs in equine plasma and urine was achieved at a concentration of 10 ng/mL.
- The validity of the method was confirmed by consistent detection of the drugs in different samples of spiked plasma and urine, with sample sizes of 15 for each type.
- Through a test done on 30 samples of blank urine, it was confirmed that there were no significant matrix interferences during the detection process. This means that the results were not impacted by any unintended substances, ensuring the accuracy of the detection.
- This method was further validated by its successful use in analysing actual drug-administration samples as well as official racing samples.
Conclusion
- The study concludes that the developed LC-MS-MS method effectively detects the presence of 10 various anti-diabetics in equine plasma and urine. It provides a robust tool to address the unfair use of these drugs in horse racing.
Cite This Article
APA
Ho EN, Yiu KC, Wan TS, Stewart BD, Watkins KL.
(2004).
Detection of anti-diabetics in equine plasma and urine by liquid chromatography-tandem mass spectrometry.
J Chromatogr B Analyt Technol Biomed Life Sci, 811(1), 65-73.
https://doi.org/10.1016/j.jchromb.2004.03.070 Publication
Researcher Affiliations
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, NT, Hong Kong, China. emmie.nm.ho@hkjc.org.hk
MeSH Terms
- Animals
- Chromatography, High Pressure Liquid / methods
- Hypoglycemic Agents / blood
- Hypoglycemic Agents / urine
- Reproducibility of Results
- Sensitivity and Specificity
- Spectrometry, Mass, Electrospray Ionization / methods
- Swine
Citations
This article has been cited 9 times.- Zhang T, Peng T, Rao J, Wang K, Qiu F. Quantitation of Diclofenac, Tolbutamide, and Warfarin as Typical CYP2C9 Substrates in Rat Plasma by UPLC-MS/MS and Its Application to Evaluate Linderane-Mediated Herb-Drug Interactions.. J Anal Methods Chem 2022;2022:1900037.
- Pamidimarri G, Mathaiyan J, Manikandan S, Rajan S, Batmanabane G. Screening of herbal medicines for potential allopathic antidiabetic adulterants: An analytical study.. Ayu 2019 Oct-Dec;40(4):262-272.
- Alam MA, Al-Jenoobi FI, Al-Mohizea AM. Rapid, Validated UPLC-MS/MS Method for Determination of Glibenclamide in Rat Plasma.. Int J Anal Chem 2018;2018:2569027.
- Polagani SR, Pilli NR, Gajula R, Gandu V. Simultaneous determination of atorvastatin, metformin and glimepiride in human plasma by LC-MS/MS and its application to a human pharmacokinetic study.. J Pharm Anal 2013 Feb;3(1):9-19.
- Pani NR, Nath L, Singh AV, Mahapatra SK. Development and validation of analytical method for the estimation of nateglinide in rabbit plasma.. J Pharm Anal 2012 Dec;2(6):492-498.
- Al-Arfaj NA, Al-Abdulkareem EA, Aly FA. A Validated Adsorptive Stripping Voltammetric Determination of Antidiabetic Agent Pioglitazone HCl in Tablets and Biological Fluids.. Int J Biomed Sci 2008 Dec;4(4):310-8.
- Kundlik ML, Zaware BH, Kuchekar SR. Rapid and specific approach for direct measurement of glimepiride in human plasma by LC-ESI-MS-MS employing automated 96 well format: application to a bioequivalence study.. J Chromatogr Sci 2012 Jan;50(1):64-70.
- Thiruvengada RV, Mohamed STs, Ramkanth S, Alagusundaram M, Ganaprakash K, Madhusudhana CC. A Simple RP-HPLC Method for Quantitation of Itopride HCl in Tablet Dosage Form.. J Young Pharm 2010 Oct;2(4):410-3.
- Hess C, Musshoff F, Madea B. Disorders of glucose metabolism-post mortem analyses in forensic cases: part I.. Int J Legal Med 2011 Mar;125(2):163-70.
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