Dataset Information

SRM-MS Method Development for Hepcidin-25 Peptide.

ABSTRACT: As advanced mass spectrometry- (MS-) based hepcidin analysis offers to overcome the limitations in analytical methods using antihepcidin, further improvement of MS detection sensitivity for the peptide may enhance the diagnostic value of the hepcidin for various iron-related disorders. Here, improved MS detection sensitivity of hepcidin has been achieved by reducing the disulfide bonds in hepcidin, by which proton accessibility increased, compared to the intact hepcidin peptide. Comparing the ionization efficiencies of reduced and nonreduced forms of hepcidin, the reduced form of hepcidin showed an increase in ionization efficiency more than two times compared to the nonreduced form of hepcidin. We also demonstrated improved detection sensitivity of the peptide in SRM assay. We observed a significant improvement of detection sensitivity at the triple-quadrupole MS platform, that the ionization efficiency increased at least twice more, and that the limit of detection (LOD) increased more than 10 times in the concentration ranges of 1?fmol to 10?fmol of hepcidin. In this study, we demonstrated the usefulness of the hepcidin modification for overall enhancement of the ionization efficiencies of the hepcidin peptide in the MS-based quantitative measurement assay.

SUBMITTER: Cho KC

PROVIDER: S-EPMC6022310 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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SRM-MS Method Development for Hepcidin-25 Peptide.

Cho Kyung-Cho KC Cho Byoung-Kyu BK Jung Jin Woo JW Lee Ye Ji YJ Lee Eun Bong EB Yi Eugene C EC

Journal of analytical methods in chemistry 20180614

As advanced mass spectrometry- (MS-) based hepcidin analysis offers to overcome the limitations in analytical methods using antihepcidin, further improvement of MS detection sensitivity for the peptide may enhance the diagnostic value of the hepcidin for various iron-related disorders. Here, improved MS detection sensitivity of hepcidin has been achieved by reducing the disulfide bonds in hepcidin, by which proton accessibility increased, compared to the intact hepcidin peptide. Comparing the io ...[more]

PMID: 30013808

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Project description:Mycotoxin contamination in animal milk is an emerging concern around the globe. Here we developed and validated an ultrahigh-performance liquid chromatography and mass spectrometry-selected reaction monitoring (UHPLC/MS-SRM) method to quantify low concentrations of aflatoxins (AFs) and ochratoxins (OTs) in routinely consumed animal milk samples collected from southern India. Stable isotope dilution methodology was applied to quantify AFB1, AFB2, AFG1, AFG2, AFM1, AFM2 and OTA, OTB in n = 38 different milk samples, using 1 mL of milk. Bioanalytical parameters including method accuracy, precision, recovery, regression analysis and stability were assessed. Dynamic ranges for quantification were between 15.6-1000 pg/mL for AFB1, AFB2, AFG1, and OTA; 7.8-500 pg/mL for AFM1, AFM2 and OTB; 78.6-5000 pg/mL for AFG2. Method accuracy ranged between 80-120%, with ± 15% precision. Recoveries for spiked standards were > 88% in water and 75% in milk, with limits of quantification (LOQ) ranging between 31.3 pg/mL for AFB1, AFB2, AFG1 and OTA, 15.6 pg/mL for AFM1, AFM2 and OTB and 156 pg/mL for AFG2. R2 values for regression analyses ranged between 0.9991-0.9999. AFB2 [mean: 38 pg/mL (0.038 µg/kg)] was quantified in goat milk, AFM1 was quantified in cow, goat, pasteurized milk [mean: 331 pg/mL (0.331 µg/kg), 406 pg/mL (0.406 µg/kg), 164 pg/mL (0.164 µg/kg)]. Additionally, 90% of cow, goat and pasteurized milk samples were above European Union (EU) limits of 50 pg/mL (0.05 µg/kg) and 40% of cow and goat milk samples were above the Food Safety Standards Authority of India (FSSAI) limit of 500 pg/mL (0.5 µg/kg). AFM2 was also quantified in cow, goat, and pasteurized milk samples [mean: 249 pg/mL (0.249 µg/kg), 375 pg/mL (0.375 µg/kg), 81 pg/mL (0.081 µg/kg)]. Our dynamic ranges for quantification are lower than other published methods, with need for a smaller volume of milk. This validated method can be applied for routine quantification of mycotoxins in milk.Supplementary informationThe online version contains supplementary material available at (10.1007/s13197-021-04986-w).

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Dataset Information

SRM-MS Method Development for Hepcidin-25 Peptide.

Publications

SRM-MS Method Development for Hepcidin-25 Peptide.

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