Orotic Acid in Milk

Hallanger, L. E.; Laakso, Joe; Schultze, Michael

doi:10.1016/s0021-9258(19)57109-5

Cited by 46 publications

(2 citation statements)

References 13 publications

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“…Moreover, the use of the very selective two C 18 -columns and OAc monitoring at the longer wavelength (i.e., 278 nm) reduced matrix effects compared to Fourier transform infrared spectral analysis [ 6 ], nuclear magnetic resonance spectroscopy [ 7 ], as well as microbiological [ 10 ], polarographic, colorimetric [ 11 ], enzymatic [ 12 ], and enzymatic-spectrophotometric [ 13 ] methods.…”

Section: Resultsmentioning

confidence: 99%

“…So far, OAc in milk and dairy products can be determined by Fourier transform infrared spectral analysis [ 4 , 6 ], nuclear magnetic resonance spectroscopy [ 7 ] and ion-chromatography [ 8 ] as well as using capillary electrophoretic [ 9 ], microbiological [ 10 ], polarographic, colorimetric [ 11 ], enzymatic [ 12 ], and enzymatic-spectrophotometric [ 13 ] methods. These methods have a very wide interval concentration range (19–664 mg OAc/L) [ 3 , 4 , 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

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The Improved Method for Determination of Orotic Acid in Milk by Ultra-Fast Liquid Chromatography with Optimized Photodiode Array Detection

Czauderna

Białek

Molik

et al. 2021

Animals

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Ultra-fast liquid chromatography (UFLC) with a photodiode array detector (DAD) for simple and rapid determination of orotic acid (OAc) in milk of sheep and cows is described. Milk samples are treated with acetonitrile (1:1, v/v) and then centrifuged at 4 °C. To 1 mL of the obtained supernatant 9 mL of ultrapure water was added. Subsequently, 0.5–6 µL of the resulting solution was injected into the UFLC-DAD system. Separation and quantification of OAc in milk samples was achieved using two Kinetex C18 columns (1.7 µm, 150 mm × 2.1 mm, i.d., 100 Å; Phenomenex) fitted with a pre-column of 4 mm × 2 mm, i.d. (Phenomenex) containing C18 packing material. All separations were performed at a column temperature of 35 °C while the ambient temperature was 21–24 °C. Satisfactory separation of OAc from endogenous species of milk can be achieved using the binary gradient elution program and UV detection at wavelengths 278 nm. Our original procedure resulted in suitable separation and quantification of OAc in milk samples; OAc eluted at 6.44 ± 0.03 min. The total run time of OAc analysis (including re-equilibration) was 27 min. As expected, the OAc peak was absent from the blank when the proposed gradient elution program and UV detection at 278 nm was used. The average recoveries of OAc standards added to milk samples were satisfactory (96.7–105.3%). The low inter-and intra-assay coefficient of variation derived from the measurements of OAc in cow and ovine milk samples (i.e., 0.784%, 1.283% and 0.710%, 1.221%, respectively) and in O-Ac standards (i.e., 0.377% and 0.294%, respectively), as well as high recoveries of OAc added to ovine and cows’ milk (~100%) and the low detection (0.04 ng) and quantification (0.12 ng) limits point to satisfactory accuracy, precision and sensitivity of the reported method. OAc concentrations in ovine milk samples were within the range from 25 to 36 mg/L, while OAc levels in cows’ milk samples was found in the range of 32–36 mg/L. Our original procedure is suitable for routine quantification of OAc in milk of ewes and cows.

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