Quantitation of vitamin K1 in serum using online SPE-LC-MS/MS and the challenges of working with vitamin K

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Resumé

Vitamin K1 (phylloquinone) is one of the vitamin Ks. Several studies have previously investigated the role of vitamin K1 status in respect to disease, but without consistent results. Since vitamin K deficiency has been associated with different disease states it is important to develop a biochemical analysis with sufficient sensitivity and a low limit of quantitation (LOQ). The vitamin Ks are very fat-soluble. This non-polar nature has given rise to several challenges during the method development, because vitamin K1 sticks to materials used during the process and is lost during evaporation. We found that reducing the sample preparation as much as possible offline, instead using online SPE-LC-MS/MS improves recovery and gives satisfactory chromatograms. An Protein BEH C4 column, 300 Å (50 × 2.1 mm, 1.7 μm particle size) was used as trap column and a Phenyl-Hexyl-LC-column, 100 Å (100 × 2.1 mm, 2.6 μm particle size) was used as analytical column. The mobile phases consisted of 30 μmol/L NH 4 F in water and 30 μmol/L NH 4 F in MeOH. A triple quadrupole tandem mass spectrometer with atmospheric pressure chemical ionization (APCI) ion source, positive ion mode, was used to perform the mass spectrometric measurements. The method is simple, highly sensitive and fast. The method was validated for vitamin K1 with good analytical performance. With a LOQ of 0.05 nmol/L it is to our knowledge the vitamin K1 method with lowest LOQ reported to date in the literature. It can easily be automated and applied in a routine diagnostic laboratory. Blood collection tubes with different additives were tested and showed no difference. Stability of vitamin K1 in serum was tested at different temperatures (−20 °C, 4 °C and in light and dark at 20 °C over a period of 30 days) and showed that vitamin K1 is light sensitive in serum even after only one day.

OriginalsprogEngelsk
TidsskriftJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Vol/bind1117
Sider (fra-til)41-48
ISSN1570-0232
DOI
StatusUdgivet - 1. jun. 2019

Fingeraftryk

Vitamin K 1
Vitamin K
Ion sources
Vitamins
Particle size
Mass spectrometers
Atmospheric pressure
Evaporation
Blood
Positive ions
Fats
Recovery
Water

Citer dette

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title = "Quantitation of vitamin K1 in serum using online SPE-LC-MS/MS and the challenges of working with vitamin K",
abstract = "Vitamin K1 (phylloquinone) is one of the vitamin Ks. Several studies have previously investigated the role of vitamin K1 status in respect to disease, but without consistent results. Since vitamin K deficiency has been associated with different disease states it is important to develop a biochemical analysis with sufficient sensitivity and a low limit of quantitation (LOQ). The vitamin Ks are very fat-soluble. This non-polar nature has given rise to several challenges during the method development, because vitamin K1 sticks to materials used during the process and is lost during evaporation. We found that reducing the sample preparation as much as possible offline, instead using online SPE-LC-MS/MS improves recovery and gives satisfactory chromatograms. An Protein BEH C4 column, 300 {\AA} (50 × 2.1 mm, 1.7 μm particle size) was used as trap column and a Phenyl-Hexyl-LC-column, 100 {\AA} (100 × 2.1 mm, 2.6 μm particle size) was used as analytical column. The mobile phases consisted of 30 μmol/L NH 4 F in water and 30 μmol/L NH 4 F in MeOH. A triple quadrupole tandem mass spectrometer with atmospheric pressure chemical ionization (APCI) ion source, positive ion mode, was used to perform the mass spectrometric measurements. The method is simple, highly sensitive and fast. The method was validated for vitamin K1 with good analytical performance. With a LOQ of 0.05 nmol/L it is to our knowledge the vitamin K1 method with lowest LOQ reported to date in the literature. It can easily be automated and applied in a routine diagnostic laboratory. Blood collection tubes with different additives were tested and showed no difference. Stability of vitamin K1 in serum was tested at different temperatures (−20 °C, 4 °C and in light and dark at 20 °C over a period of 30 days) and showed that vitamin K1 is light sensitive in serum even after only one day.",
author = "{Boegh Andersen}, Ida and {Lohman Brasen}, Claus and {Skov Madsen}, Jonna and Anne Schmedes",
year = "2019",
month = "6",
day = "1",
doi = "10.1016/j.jchromb.2019.04.010",
language = "English",
volume = "1117",
pages = "41--48",
journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences",
issn = "1570-0232",
publisher = "Elsevier",

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TY - JOUR

T1 - Quantitation of vitamin K1 in serum using online SPE-LC-MS/MS and the challenges of working with vitamin K

AU - Boegh Andersen, Ida

AU - Lohman Brasen, Claus

AU - Skov Madsen, Jonna

AU - Schmedes, Anne

PY - 2019/6/1

Y1 - 2019/6/1

N2 - Vitamin K1 (phylloquinone) is one of the vitamin Ks. Several studies have previously investigated the role of vitamin K1 status in respect to disease, but without consistent results. Since vitamin K deficiency has been associated with different disease states it is important to develop a biochemical analysis with sufficient sensitivity and a low limit of quantitation (LOQ). The vitamin Ks are very fat-soluble. This non-polar nature has given rise to several challenges during the method development, because vitamin K1 sticks to materials used during the process and is lost during evaporation. We found that reducing the sample preparation as much as possible offline, instead using online SPE-LC-MS/MS improves recovery and gives satisfactory chromatograms. An Protein BEH C4 column, 300 Å (50 × 2.1 mm, 1.7 μm particle size) was used as trap column and a Phenyl-Hexyl-LC-column, 100 Å (100 × 2.1 mm, 2.6 μm particle size) was used as analytical column. The mobile phases consisted of 30 μmol/L NH 4 F in water and 30 μmol/L NH 4 F in MeOH. A triple quadrupole tandem mass spectrometer with atmospheric pressure chemical ionization (APCI) ion source, positive ion mode, was used to perform the mass spectrometric measurements. The method is simple, highly sensitive and fast. The method was validated for vitamin K1 with good analytical performance. With a LOQ of 0.05 nmol/L it is to our knowledge the vitamin K1 method with lowest LOQ reported to date in the literature. It can easily be automated and applied in a routine diagnostic laboratory. Blood collection tubes with different additives were tested and showed no difference. Stability of vitamin K1 in serum was tested at different temperatures (−20 °C, 4 °C and in light and dark at 20 °C over a period of 30 days) and showed that vitamin K1 is light sensitive in serum even after only one day.

AB - Vitamin K1 (phylloquinone) is one of the vitamin Ks. Several studies have previously investigated the role of vitamin K1 status in respect to disease, but without consistent results. Since vitamin K deficiency has been associated with different disease states it is important to develop a biochemical analysis with sufficient sensitivity and a low limit of quantitation (LOQ). The vitamin Ks are very fat-soluble. This non-polar nature has given rise to several challenges during the method development, because vitamin K1 sticks to materials used during the process and is lost during evaporation. We found that reducing the sample preparation as much as possible offline, instead using online SPE-LC-MS/MS improves recovery and gives satisfactory chromatograms. An Protein BEH C4 column, 300 Å (50 × 2.1 mm, 1.7 μm particle size) was used as trap column and a Phenyl-Hexyl-LC-column, 100 Å (100 × 2.1 mm, 2.6 μm particle size) was used as analytical column. The mobile phases consisted of 30 μmol/L NH 4 F in water and 30 μmol/L NH 4 F in MeOH. A triple quadrupole tandem mass spectrometer with atmospheric pressure chemical ionization (APCI) ion source, positive ion mode, was used to perform the mass spectrometric measurements. The method is simple, highly sensitive and fast. The method was validated for vitamin K1 with good analytical performance. With a LOQ of 0.05 nmol/L it is to our knowledge the vitamin K1 method with lowest LOQ reported to date in the literature. It can easily be automated and applied in a routine diagnostic laboratory. Blood collection tubes with different additives were tested and showed no difference. Stability of vitamin K1 in serum was tested at different temperatures (−20 °C, 4 °C and in light and dark at 20 °C over a period of 30 days) and showed that vitamin K1 is light sensitive in serum even after only one day.

U2 - 10.1016/j.jchromb.2019.04.010

DO - 10.1016/j.jchromb.2019.04.010

M3 - Journal article

VL - 1117

SP - 41

EP - 48

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SN - 1570-0232

ER -