Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard
An ultrahigh-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the determination of lactoferrin in camel milk based on the signature peptide. The camel lactoferrin was purified by heparin affinity chromatography and then used to screen tryptic signatur...
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MDPI AG
2019-11-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/24/22/4199 |
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| author | Xia Li Zengmei Li Enmin Xu Ling Chen Hua Feng Lu Chen Ligang Deng Dongliang Guo |
| author_facet | Xia Li Zengmei Li Enmin Xu Ling Chen Hua Feng Lu Chen Ligang Deng Dongliang Guo |
| author_sort | Xia Li |
| collection | DOAJ |
| description | An ultrahigh-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the determination of lactoferrin in camel milk based on the signature peptide. The camel lactoferrin was purified by heparin affinity chromatography and then used to screen tryptic signature peptides. The signature peptide was selected on the basis of sequence database search and identified from the tryptic hydrolysates of purified camel lactoferrin by ultrahigh-performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry. The pretreatment procedures included the addition of isotope-labeled winged peptide and the disposal of lipids and caseins followed by an enzymatic digestion with trypsin. Analytes were separated on an Acquity UPLC BEH 300 C18 column and then detected on a triple-quadrupole mass spectrometer in 7 min. The limits of detection and quantification were 3.8 mg kg<sup>−1</sup> and 11 mg kg<sup>−1</sup>, respectively. The recoveries ranged from 74.5% to 103.6%, with relative standard deviations below 7.7%. The validated method was applied to determine the lactoferrin in ten samples collected from Xinjiang Province. |
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| language | English |
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| publishDate | 2019-11-01 |
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| spelling | doaj.art-5b77fdb22a2f4187b12e0675d5b38a262022-12-22T00:09:47ZengMDPI AGMolecules1420-30492019-11-012422419910.3390/molecules24224199molecules24224199Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal StandardXia Li0Zengmei Li1Enmin Xu2Ling Chen3Hua Feng4Lu Chen5Ligang Deng6Dongliang Guo7Institute of Agricultural Quality Standards and Testing Technology Research, Shandong Academy of Agricultural Sciences, 202 Gongyebeilu Road, Jinan 250100, Shandong Province, ChinaInstitute of Agricultural Quality Standards and Testing Technology Research, Shandong Academy of Agricultural Sciences, 202 Gongyebeilu Road, Jinan 250100, Shandong Province, ChinaShandong Veterinary Drug Quality Inspection Institute, 68 Huaicun Street, Jinan, Shandong Province, ChinaShandong Veterinary Drug Quality Inspection Institute, 68 Huaicun Street, Jinan, Shandong Province, ChinaInstitute of Agricultural Quality Standards and Testing Technology Research, Shandong Academy of Agricultural Sciences, 202 Gongyebeilu Road, Jinan 250100, Shandong Province, ChinaInstitute of Agricultural Quality Standards and Testing Technology Research, Shandong Academy of Agricultural Sciences, 202 Gongyebeilu Road, Jinan 250100, Shandong Province, ChinaInstitute of Agricultural Quality Standards and Testing Technology Research, Shandong Academy of Agricultural Sciences, 202 Gongyebeilu Road, Jinan 250100, Shandong Province, ChinaInstitute of Agricultural Quality Standards and Testing Technology Research, Shandong Academy of Agricultural Sciences, 202 Gongyebeilu Road, Jinan 250100, Shandong Province, ChinaAn ultrahigh-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the determination of lactoferrin in camel milk based on the signature peptide. The camel lactoferrin was purified by heparin affinity chromatography and then used to screen tryptic signature peptides. The signature peptide was selected on the basis of sequence database search and identified from the tryptic hydrolysates of purified camel lactoferrin by ultrahigh-performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry. The pretreatment procedures included the addition of isotope-labeled winged peptide and the disposal of lipids and caseins followed by an enzymatic digestion with trypsin. Analytes were separated on an Acquity UPLC BEH 300 C18 column and then detected on a triple-quadrupole mass spectrometer in 7 min. The limits of detection and quantification were 3.8 mg kg<sup>−1</sup> and 11 mg kg<sup>−1</sup>, respectively. The recoveries ranged from 74.5% to 103.6%, with relative standard deviations below 7.7%. The validated method was applied to determine the lactoferrin in ten samples collected from Xinjiang Province.https://www.mdpi.com/1420-3049/24/22/4199lactoferrincamel milkultrahigh-performance liquid chromatography-tandem mass spectrometryheparin affinity chromatography |
| spellingShingle | Xia Li Zengmei Li Enmin Xu Ling Chen Hua Feng Lu Chen Ligang Deng Dongliang Guo Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard Molecules lactoferrin camel milk ultrahigh-performance liquid chromatography-tandem mass spectrometry heparin affinity chromatography |
| title | Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard |
| title_full | Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard |
| title_fullStr | Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard |
| title_full_unstemmed | Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard |
| title_short | Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard |
| title_sort | determination of lactoferrin in camel milk by ultrahigh performance liquid chromatography tandem mass spectrometry using an isotope labeled winged peptide as internal standard |
| topic | lactoferrin camel milk ultrahigh-performance liquid chromatography-tandem mass spectrometry heparin affinity chromatography |
| url | https://www.mdpi.com/1420-3049/24/22/4199 |
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