Determination of vitamin B12 in equine urine by liquid chromatography – inductively coupled plasma – mass spectrometry

Wenzel, Ross; Major, Derek; Hesp, Karly; Doble, Philip

doi:10.1016/j.jtemb.2018.05.005

Cited by 8 publications

(4 citation statements)

References 15 publications

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“…The presented data indicate the importance of the careful assessment of total urinary cobalt concentrations in the context of routine sports drug testing programs and, potentially, also in other clinical applications. The determination of inorganic cobalt and the differentiation from cyanocobalamin can be accommodated by simple SPE‐based depletion of the cobalt‐containing organic molecule and subsequent analysis of the extract's flow‐through by ICP‐MS.…”

mentioning

confidence: 86%

Elevated urinary cobalt concentrations identified in routine doping controls can originate from vitamin B₁₂

Knoop

Görgens

Geyer

et al. 2020

Rapid Comm Mass Spectrometry

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mentioning

confidence: 86%

Elevated urinary cobalt concentrations identified in routine doping controls can originate from vitamin B₁₂

Knoop

Görgens

Geyer

et al. 2020

Rapid Comm Mass Spectrometry

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“…e existing reported methods for determination of VB 12 in infant formula mainly include microbiological assays [9,10], high-performance liquid chromatography (HPLC) [11][12][13][14][15][16][17][18][19], liquid chromatography-tandem mass spectrometry (LC-MS/MS) [20][21][22][23][24][25][26], and liquid chromatography and inductively coupled plasma-mass spectrometry (LC-ICP-MS) [27][28][29]. e microbiological assays usually have a long measurement time and complicated operation.…”

Section: Introductionmentioning

confidence: 99%

Determination of Vitamin B12 in Milk and Dairy Products by Isotope-Dilution Liquid Chromatography Tandem Mass Spectrometry

Huang

Zhang

Cai

2022

Journal of Food Quality

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An isotope-dilution liquid chromatography tandem mass spectrometry method was established for the determination of vitamin B12 in milk and dairy products. The samples were spiked with stable isotope-labeled vitamin B12 and digested by pepsin and amylase. The various forms of cobalamin were transformed to cyanocobalamin by potassium cyanide after they were released from the enzymatically digested samples. Cyanocobalamin was extracted and purified by an immunoaffinity SPE cartridge and then measured in multiple reaction monitoring mode (MRM). The linear correlation coefficient (R2) of this method was greater than 0.999 in the range of 2–100 ng/mL. The detection limit and the quantification limit were 0.5 μg/kg and 1.0 μg/kg, respectively. The spiked recoveries ranged from 92.0% to 99.4% at the three spiked levels with the relative standard deviation (RSD) between 1.89% and 4.51%. The measured results of NIST SRM1849a and NIST SRM1869a by the current method are all within the reference value range. The Z value was 0.8 during participating in the FAPAS proficiency test using the developed method in 2021. The method is simple, rapid, accurate, and sensitive, and it is suitable for the determination of vitamin B12 in different types of milk and dairy products such as whey powder, whole milk powder, pure milk, fermented milk, infant formula, and prescription food for special medical purposes.

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“…With regard to the scope of doping controls, Popot et al 20 presented interlaboratory data (ring tests) for urine und plasma measurements of total Co by ICP‐MS for equestrian sports. Furthermore, the International Federation of Horseracing Authorities (IFHA) have set a threshold value of 100 μg Co per liter of urine 21 …”

Section: Introductionmentioning

confidence: 99%

“…Excretion studies with either CoCl 2 or cyanocobalamin were conducted with two separate groups of healthy volunteers, and urine as well as EDTA whole blood samples were frequently collected over a period of 30 days. In this proof‐of‐concept study, a test method enabling the separation and detection of cobalt‐containing organic analytes and unbound cobalt in human urine within one single analytical run was adapted as a simplified screening from former publications 21–24 utilizing high performance liquid chromatography (HPLC) combined with ICP‐MS. The approach aims at facilitating the immediate attribution of measured urinary cobalt concentrations to either the legitimate use of cyanocobalamin or the prohibited use of, for example, cobalt salts.…”

Section: Introductionmentioning

confidence: 99%

Analysis of cobalt for human sports drug testing purposes using ICP‐ and LC‐ICP‐MS

Knoop

Planitz

Wüst

et al. 2020

Drug Testing and Analysis

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Due to the current demands in the fight against manipulation of blood and blood components, commonly referred to as “blood doping” in sports drug testing, specific and sensitive detection methods enabling the detection of prohibited substances and methods of doping are required. Similar to illicit blood transfusions, erythropoiesis stimulating agents have been shown to be misused in sport, aiming at improving an athlete's aerobic capacity and endurance performance. Amongst other strategies, the administration of ionic cobalt (Co2+) can increase the number of erythrocytes by stimulating the endogenous erythropoietin (EPO) biosynthesis. Conversely, several organic Co‐containing compounds such as cyanocobalamin (vitamin B12) are not prohibited in sports, and thus, an analytical differentiation of permitted and banned contributions to urinary Co‐concentrations is desirable. An excretion study with daily applications of either 1 mg of CoCl2 or 1 mg of cyanocobalamin was conducted with 20 volunteers over a period of 14 consecutive days. Urine, plasma, and concentrated red blood cells were analyzed for their cobalt content. The samples were collected starting 7 days before the administration until 7 days after. Total Co concentrations were analyzed by using inductively coupled plasma mass spectrometry (ICP‐MS), which yielded significantly elevated levels exclusively after inorganic cobalt intake. Furthermore, a liquid chromatography (LC)‐ICP‐MS approach was established and employed for the simultaneous determination of organically bound and inorganic cobalt by chromatographic separation within one single run. The analytical approach offers the option to further develop detection methods of illegal Co2+ supplementation in sport.

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Determination of vitamin B12 in equine urine by liquid chromatography – inductively coupled plasma – mass spectrometry

Cited by 8 publications

References 15 publications

Elevated urinary cobalt concentrations identified in routine doping controls can originate from vitamin B₁₂

Elevated urinary cobalt concentrations identified in routine doping controls can originate from vitamin B₁₂

Determination of Vitamin B12 in Milk and Dairy Products by Isotope-Dilution Liquid Chromatography Tandem Mass Spectrometry

Analysis of cobalt for human sports drug testing purposes using ICP‐ and LC‐ICP‐MS

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Determination of vitamin B12 in equine urine by liquid chromatography – inductively coupled plasma – mass spectrometry

Cited by 8 publications

References 15 publications

Elevated urinary cobalt concentrations identified in routine doping controls can originate from vitamin B12

Elevated urinary cobalt concentrations identified in routine doping controls can originate from vitamin B12

Determination of Vitamin B12 in Milk and Dairy Products by Isotope-Dilution Liquid Chromatography Tandem Mass Spectrometry

Analysis of cobalt for human sports drug testing purposes using ICP‐ and LC‐ICP‐MS

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Elevated urinary cobalt concentrations identified in routine doping controls can originate from vitamin B₁₂

Elevated urinary cobalt concentrations identified in routine doping controls can originate from vitamin B₁₂