Separation of Tocols by HPLC on an Amino-Cyano Polar Phase Column

Rammell, C. G.; Hoogenboom, J.J.L.

doi:10.1080/01483918508067114

Cited by 30 publications

(14 citation statements)

References 13 publications

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“…α-Tocopherol is absent in sesame oil and δ-tocopherol in olive oil. The results obtained are generally in agreement with the literature data (Berasategi et al, 2012;Rammell and Hoogenboom, 1985;Sanchez-Perez et al, 2000). Lack of separation of β-and γ-tocopherols in the case of edible oils introduce rather small error in quantification of these isomers (Gliszczynska-Swiglo and Sikorska, 2004).…”

Section: Quantification Of Tocopherols In Edible Plant Oilssupporting

confidence: 89%

Tocopherol Content in Vegetable Oils Using a Rapid HPLC Fluorescence Detection Method

Bele

Matea

Răducu

et al. 2013

Not Bot Hort Agrobot Cluj

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A quick and direct method based on reversed phase high performance liquid chromatography with fluorescence detector for measuring tocopherols (α , β + γ and δ) has been developed. Oils are diluted in methanol: hexane: tetrahydrofuran (neither previous extraction of tocopherols nor saponification procedure are required) and after being vortexed and centrifuged, an aliquot of the overlay was injected directly into an Alltima C 18 column. Acetonitrile and methanol (50: 50) mixture was used as a mobile phase with a flow rate of 1 mL min -1 . Quantification of tocopherols was performed by fluorescence detector at 290 nm excitation wavelength and 325 nm emission wavelength. Tocopherols were separated at 25°C in less than 10 min after injection. The method has good limit of detection (9 ng g -1 for α-tocopherol and 8 ng g -1 for β-, γ-and δ-tocopherols) and reproducibility (CV< 2.9 %). This method can be used to assess the influence of genetic modification of oil seeds on the distribution of tocopherols or the effect of tocopherols on the oxidative stability of edible plant oils.

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Section: Quantification Of Tocopherols In Edible Plant Oilssupporting

confidence: 89%

Tocopherol Content in Vegetable Oils Using a Rapid HPLC Fluorescence Detection Method

Bele

Matea

Răducu

et al. 2013

Not Bot Hort Agrobot Cluj

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“…Plain tubes were kept at room temperature for 6 hours, then centrifuged and the serum decanted. Serum was kept for up to 36 hours at 4 DC until the assays of selenium(20), a-tocopherol (21) and fatty acids (22)(23) were performed. Peroxidisable polyunsaturated fatty acids were defined as all detectable polyunsaturated fatty acids, with the exception of linoleic acid, which has been shown to be relatively resistant to peroxidation (24).…”

Section: Sampling and Assaysmentioning

confidence: 99%

Alpha-tocopherol, selenium and polyunsaturated fatty acid concentrations in the serum and feed of spring-calving dairy heifers

Wichtel

Freeman

Craigie

et al. 1996

New Zealand Veterinary Journal

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The objectives of this study were to provide baseline data for alpha-tocopherol, selenium and polyunsaturated fatty acid concentrations in the serum and feed of New Zealand dairy cattle, and to assess the likelihood that abnormal peroxide metabolism has a role in the impaired lactational and reproductive performance noted in selenium-deficient cattle. Twenty-four Friesian heifers were randomly allocated one of four winter diets consisting of hay with or without selenium supplementation, or pasture and silage with or without selenium supplementation. A winter diet consisting exclusively of hay (alpha-tocopherol concentration 19 mg/kg of dry matter) resulted in a pre-calving serum alpha-tocopherol concentration of 1.2 mg/l compared to 4.5 mg/l for pastured heifers (p<0.01). The pre-calving alpha-tocopherol concentration for the heifers fed hay fell into the range considered deficient (<2.0 mg/l), whereas heifers fed pasture and silage remained in the range considered adequate throughout the study period. Serum fatty acid concentration, and the proportion of fatty acids that were polyunsaturated, were lowest in the hay-fed heifers before calving (1.0 mg/ml, 37.1% respectively), and remained unchanged following re-introduction to pasture after calving in late July and August. Serum fatty acid concentration did not increase following the re-introduction of the heifers to pasture because of the unexpectedly low fatty acid concentration (4.8 g/kg of dry matter) of the mature winter pasture. In October, however, the proportion of fatty acids in serum that were polyunsaturated increased (50%) as did serum cy-tocopherol concentrations (greater than 13 mg/l). Mean serum selenium concentrations in the unsupplemented heifers ranged from 139 to 204 nmol/l, being lowest in October (p<0.01). Supplementation with intraruminal selenium pellets (two pellets delivering about 3 mg of selenium/day) increased serum selenium concentration and glutathione peroxidase activity (p<0.01) whereas the type of winter diet had no effect (p>0.05). These results suggest that dairy cattle wintered on hay can become Vitamin E-depleted, whereas the feeding of pasture and silage should provide adequate Vitamin E. The pasture offered following calving during July and August provided a low dietary polyunsaturated fatty acid challenge, suggesting that abnormal peroxide metabolism is unlikely to be an important mechanism in the impaired performance of selenium-deficient adult cattle which calve at this the of year.

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“…The analyses of tocopherol in foods and feeds (1,8,9,14,15,(17)(18)(19)(20), oils and seeds (1,19,(21)(22)(23)(24)(25)(26)(27), plasma (2,4,6-10, 13-15,28-32), platelets (30), and red cells (4,7,28,32) are the same as for tissues, except for the prior homogenization step. Foods or feeds are ground and extracted directly (1,9) or saponified and extracted (1,8,14,15,(17)(18)(19)(20), vegetable oils are directly extracted (21)(22)(23)(24)(25)27) or saponified and then extracted (19,25,26), and plasma, platelets and red cells are treated with ethanol (2,(6)(7)(8)(9)(10)...…”

mentioning

confidence: 99%

“…The tocopherols are separated by high-performance liquid chromatography (HPLC) on normal-phase (1,(3)(4)(5)(8)(9)(10)(11)15,18,19,(21)(22)(23)(25)(26)(27), or reversed-phase columns (2,6-8, 12-14,17,19,20,24,28-32). A normal-phase column, bonded with amino-cyano groups, was used successfully to separate all vitamin E forms (9,19,23). Generally, tocopherols and tocotrienols are detected by fluorescence, although ultraviolet (3,13,19,28) or evaporative light-scattering detection (24,25) has also been used.…”

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confidence: 99%

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A rapid method for the determination of vitamin E forms in tissues and diet by high‐performance liquid chromatography using a normal‐phase diol column

Kramer¹,

Blais²,

Fouchard³

et al. 1997

Lipids

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This paper describes a simple method for the analysis of tocopherols in tissues by which frozen tissues-70 degrees C were pulverized at dry ice temperatures (-70 degrees C) and immediately extracted with hexane. There was no need to remove the coeluting lipids from tissues by saponification, since at that level of neutral lipids in the sample, there was no reduction in fluorescence response. For the analysis of oil, in which large amounts of neutral lipids were coextracted, a 20% reduction of fluorescence response was observed, but the response was equal for all tocopherol forms, and was appropriately corrected. Saponification was used only when tocopherol esters were present, and only after an initial hexane extraction to remove the free tocopherols in order to avoid their loss by saponification, particularly non alpha-tocopherol and tocotrienols. All the tocopherols and tocotrienols were separated on a normal-phase diol (epoxide) column that gave consistent and reproducible results, without the disadvantages of nonreproducibility with silica columns, or the lack of separation with reversed-phase columns. The tocopherols were quantitated by using a tocopherol form not present in the sample as an internal tocopherol standard, or using an external tocopherol standard if all forms were present, or when the sample was saponified. Piglet heart and liver samples showed the presence of mainly alpha-tocopherol, with minor amounts of beta- and gamma-tocopherol and alpha-tocotrienol, but no delta-tocopherol. Only small amounts of tocopherol esters were present in the liver but not in the heart.

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Separation of Tocols by HPLC on an Amino-Cyano Polar Phase Column

Cited by 30 publications

References 13 publications

Tocopherol Content in Vegetable Oils Using a Rapid HPLC Fluorescence Detection Method

Tocopherol Content in Vegetable Oils Using a Rapid HPLC Fluorescence Detection Method

Alpha-tocopherol, selenium and polyunsaturated fatty acid concentrations in the serum and feed of spring-calving dairy heifers

A rapid method for the determination of vitamin E forms in tissues and diet by high‐performance liquid chromatography using a normal‐phase diol column

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