CN111650286A - Method for detecting medium-long chain fatty acid in human serum based on gas chromatography-mass spectrometry - Google Patents
Method for detecting medium-long chain fatty acid in human serum based on gas chromatography-mass spectrometry Download PDFInfo
- Publication number
- CN111650286A CN111650286A CN202010252106.6A CN202010252106A CN111650286A CN 111650286 A CN111650286 A CN 111650286A CN 202010252106 A CN202010252106 A CN 202010252106A CN 111650286 A CN111650286 A CN 111650286A
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- Prior art keywords
- methyl
- chain fatty
- long chain
- mass spectrometry
- medium
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- Pending
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- 238000000034 method Methods 0.000 title claims abstract description 29
- 210000002966 serum Anatomy 0.000 title claims abstract description 25
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 title claims abstract description 21
- 241000282414 Homo sapiens Species 0.000 title claims abstract description 19
- BDXAHSJUDUZLDU-UHFFFAOYSA-N methyl nonadecanoate Chemical compound CCCCCCCCCCCCCCCCCCC(=O)OC BDXAHSJUDUZLDU-UHFFFAOYSA-N 0.000 claims abstract description 28
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 22
- 230000032050 esterification Effects 0.000 claims abstract description 5
- 238000005886 esterification reaction Methods 0.000 claims abstract description 5
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 36
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 28
- 239000000126 substance Substances 0.000 claims description 22
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 18
- 230000010355 oscillation Effects 0.000 claims description 17
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 16
- 239000011259 mixed solution Substances 0.000 claims description 16
- 238000000605 extraction Methods 0.000 claims description 11
- 150000004702 methyl esters Chemical class 0.000 claims description 11
- 239000012086 standard solution Substances 0.000 claims description 10
- 238000004949 mass spectrometry Methods 0.000 claims description 9
- 238000001819 mass spectrum Methods 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 8
- -1 methyl eicosenoate Chemical compound 0.000 claims description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 239000001149 (9Z,12Z)-octadeca-9,12-dienoate Substances 0.000 claims description 7
- FLIACVVOZYBSBS-UHFFFAOYSA-N Methyl palmitate Chemical compound CCCCCCCCCCCCCCCC(=O)OC FLIACVVOZYBSBS-UHFFFAOYSA-N 0.000 claims description 7
- HPEUJPJOZXNMSJ-UHFFFAOYSA-N Methyl stearate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC HPEUJPJOZXNMSJ-UHFFFAOYSA-N 0.000 claims description 7
- PTFHIRHGARALFY-JEBPEJKESA-N methyl (7z,10z,13z,16z,19z)-docosa-7,10,13,16,19-pentaenoate Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCCCC(=O)OC PTFHIRHGARALFY-JEBPEJKESA-N 0.000 claims description 7
- YRHYCMZPEVDGFQ-UHFFFAOYSA-N methyl decanoate Chemical compound CCCCCCCCCC(=O)OC YRHYCMZPEVDGFQ-UHFFFAOYSA-N 0.000 claims description 7
- HUEBIMLTDXKIPR-UHFFFAOYSA-N methyl heptadecanoate Chemical compound CCCCCCCCCCCCCCCCC(=O)OC HUEBIMLTDXKIPR-UHFFFAOYSA-N 0.000 claims description 7
- NUKZAGXMHTUAFE-UHFFFAOYSA-N methyl hexanoate Chemical compound CCCCCC(=O)OC NUKZAGXMHTUAFE-UHFFFAOYSA-N 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000004817 gas chromatography Methods 0.000 claims description 6
- QGBRLVONZXHAKJ-UHFFFAOYSA-N methyl arachidate Chemical compound CCCCCCCCCCCCCCCCCCCC(=O)OC QGBRLVONZXHAKJ-UHFFFAOYSA-N 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- 150000002500 ions Chemical class 0.000 claims description 5
- QSQLTHHMFHEFIY-UHFFFAOYSA-N methyl behenate Chemical compound CCCCCCCCCCCCCCCCCCCCCC(=O)OC QSQLTHHMFHEFIY-UHFFFAOYSA-N 0.000 claims description 5
- AJRICDSAJQHDSD-UHFFFAOYSA-N methyl henicosanoate Chemical compound CCCCCCCCCCCCCCCCCCCCC(=O)OC AJRICDSAJQHDSD-UHFFFAOYSA-N 0.000 claims description 5
- UQDUPQYQJKYHQI-UHFFFAOYSA-N methyl laurate Chemical compound CCCCCCCCCCCC(=O)OC UQDUPQYQJKYHQI-UHFFFAOYSA-N 0.000 claims description 5
- JGHZJRVDZXSNKQ-UHFFFAOYSA-N methyl octanoate Chemical compound CCCCCCCC(=O)OC JGHZJRVDZXSNKQ-UHFFFAOYSA-N 0.000 claims description 5
- ZAZKJZBWRNNLDS-UHFFFAOYSA-N methyl tetradecanoate Chemical compound CCCCCCCCCCCCCC(=O)OC ZAZKJZBWRNNLDS-UHFFFAOYSA-N 0.000 claims description 5
- JNDDPBOKWCBQSM-UHFFFAOYSA-N methyl tridecanoate Chemical compound CCCCCCCCCCCCC(=O)OC JNDDPBOKWCBQSM-UHFFFAOYSA-N 0.000 claims description 5
- XPQPWPZFBULGKT-UHFFFAOYSA-N methyl undecanoate Chemical compound CCCCCCCCCCC(=O)OC XPQPWPZFBULGKT-UHFFFAOYSA-N 0.000 claims description 5
- XIUXKAZJZFLLDQ-UHFFFAOYSA-N n-pentadecanoic acid methyl ester Natural products CCCCCCCCCCCCCCC(=O)OC XIUXKAZJZFLLDQ-UHFFFAOYSA-N 0.000 claims description 5
- CASUWPDYGGAUQV-UHFFFAOYSA-M potassium;methanol;hydroxide Chemical compound [OH-].[K+].OC CASUWPDYGGAUQV-UHFFFAOYSA-M 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 5
- 238000012360 testing method Methods 0.000 claims description 5
- WTTJVINHCBCLGX-UHFFFAOYSA-N (9trans,12cis)-methyl linoleate Natural products CCCCCC=CCC=CCCCCCCCC(=O)OC WTTJVINHCBCLGX-UHFFFAOYSA-N 0.000 claims description 4
- DVWSXZIHSUZZKJ-UHFFFAOYSA-N 18:3n-3 Natural products CCC=CCC=CCC=CCCCCCCCC(=O)OC DVWSXZIHSUZZKJ-UHFFFAOYSA-N 0.000 claims description 4
- UZEPQZBOMGDJGU-UHFFFAOYSA-N 2-methyldocos-2-enoic acid Chemical compound CCCCCCCCCCCCCCCCCCCC=C(C)C(O)=O UZEPQZBOMGDJGU-UHFFFAOYSA-N 0.000 claims description 4
- LNJCGNRKWOHFFV-UHFFFAOYSA-N 3-(2-hydroxyethylsulfanyl)propanenitrile Chemical compound OCCSCCC#N LNJCGNRKWOHFFV-UHFFFAOYSA-N 0.000 claims description 4
- VVAKRCKTAVIPGG-UHFFFAOYSA-N CCCCCCCCCCCCCCCCCCCCCC=C(C)C(O)=O Chemical compound CCCCCCCCCCCCCCCCCCCCCC=C(C)C(O)=O VVAKRCKTAVIPGG-UHFFFAOYSA-N 0.000 claims description 4
- PKIXXJPMNDDDOS-UHFFFAOYSA-N Methyl linoleate Natural products CCCCC=CCCC=CCCCCCCCC(=O)OC PKIXXJPMNDDDOS-UHFFFAOYSA-N 0.000 claims description 4
- QWDCYFDDFPWISL-JEBPEJKESA-N cis-5,8,11,14,17-eicosapentaenoic acid methyl ester Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(=O)OC QWDCYFDDFPWISL-JEBPEJKESA-N 0.000 claims description 4
- QYDYPVFESGNLHU-UHFFFAOYSA-N elaidic acid methyl ester Natural products CCCCCCCCC=CCCCCCCCC(=O)OC QYDYPVFESGNLHU-UHFFFAOYSA-N 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 4
- ZYNDJIBBPLNPOW-KHPPLWFESA-N methyl erucate Chemical compound CCCCCCCC\C=C/CCCCCCCCCCCC(=O)OC ZYNDJIBBPLNPOW-KHPPLWFESA-N 0.000 claims description 4
- DVWSXZIHSUZZKJ-YSTUJMKBSA-N methyl linolenate Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(=O)OC DVWSXZIHSUZZKJ-YSTUJMKBSA-N 0.000 claims description 4
- QYDYPVFESGNLHU-KHPPLWFESA-N methyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC QYDYPVFESGNLHU-KHPPLWFESA-N 0.000 claims description 4
- UUIQMZJEGPQKFD-UHFFFAOYSA-N n-butyric acid methyl ester Natural products CCCC(=O)OC UUIQMZJEGPQKFD-UHFFFAOYSA-N 0.000 claims description 4
- JGFBQFKZKSSODQ-UHFFFAOYSA-N Isothiocyanatocyclopropane Chemical compound S=C=NC1CC1 JGFBQFKZKSSODQ-UHFFFAOYSA-N 0.000 claims description 3
- 239000005640 Methyl decanoate Substances 0.000 claims description 3
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 3
- OFIDNKMQBYGNIW-UHFFFAOYSA-N arachidonic acid methyl ester Natural products CCCCCC=CCC=CCC=CCC=CCCCC(=O)OC OFIDNKMQBYGNIW-UHFFFAOYSA-N 0.000 claims description 3
- PWLNAUNEAKQYLH-UHFFFAOYSA-N butyric acid octyl ester Natural products CCCCCCCCOC(=O)CCC PWLNAUNEAKQYLH-UHFFFAOYSA-N 0.000 claims description 3
- CAMHHLOGFDZBBG-UHFFFAOYSA-N epoxidized methyl oleate Natural products CCCCCCCCC1OC1CCCCCCCC(=O)OC CAMHHLOGFDZBBG-UHFFFAOYSA-N 0.000 claims description 3
- ZYNDJIBBPLNPOW-UHFFFAOYSA-N eurucic acid methyl ester Natural products CCCCCCCCC=CCCCCCCCCCCCC(=O)OC ZYNDJIBBPLNPOW-UHFFFAOYSA-N 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 229940073769 methyl oleate Drugs 0.000 claims description 3
- IZFGRAGOVZCUFB-HJWRWDBZSA-N methyl palmitoleate Chemical compound CCCCCC\C=C/CCCCCCCC(=O)OC IZFGRAGOVZCUFB-HJWRWDBZSA-N 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000012545 processing Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- ZEBIJPXNSBSHGD-ZKWNWVNESA-N (5z,8z,11z,14z)-2-methylicosa-5,8,11,14-tetraenoic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCC(C)C(O)=O ZEBIJPXNSBSHGD-ZKWNWVNESA-N 0.000 claims description 2
- XLQNWWNMESYKTB-UHFFFAOYSA-N 2-fluoro-1h-benzimidazole Chemical compound C1=CC=C2NC(F)=NC2=C1 XLQNWWNMESYKTB-UHFFFAOYSA-N 0.000 claims description 2
- 239000005641 Methyl octanoate Substances 0.000 claims description 2
- QWDCYFDDFPWISL-UHFFFAOYSA-N UNPD207407 Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(=O)OC QWDCYFDDFPWISL-UHFFFAOYSA-N 0.000 claims description 2
- 150000004668 long chain fatty acids Chemical class 0.000 claims description 2
- VTDHQZRGOOTGNR-UHFFFAOYSA-N methyl docosa-2,4,6,8-tetraenoate Chemical compound CCCCCCCCCCCCCC=CC=CC=CC=CC(=O)OC VTDHQZRGOOTGNR-UHFFFAOYSA-N 0.000 claims description 2
- UBFGEEJWHSVIER-UHFFFAOYSA-N methyl heptadec-2-enoate Chemical compound CCCCCCCCCCCCCCC=CC(=O)OC UBFGEEJWHSVIER-UHFFFAOYSA-N 0.000 claims description 2
- WQCYAHKAJFZVCO-UHFFFAOYSA-N omega-Oxy-pentadecylsaeure-methylester Natural products COC(=O)CCCCCCCCCCCCCCO WQCYAHKAJFZVCO-UHFFFAOYSA-N 0.000 claims description 2
- 150000004667 medium chain fatty acids Chemical class 0.000 claims 2
- 238000007664 blowing Methods 0.000 claims 1
- 238000004458 analytical method Methods 0.000 abstract description 3
- 238000000622 liquid--liquid extraction Methods 0.000 abstract description 3
- 238000000638 solvent extraction Methods 0.000 abstract description 3
- 238000004587 chromatography analysis Methods 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 238000004885 tandem mass spectrometry Methods 0.000 abstract description 2
- QYDYPVFESGNLHU-ZHACJKMWSA-N Methyl (9E)-9-octadecenoate Chemical compound CCCCCCCC\C=C\CCCCCCCC(=O)OC QYDYPVFESGNLHU-ZHACJKMWSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- XUDJZDNUVZHSKZ-UHFFFAOYSA-N methyl tetracosanoate Chemical compound CCCCCCCCCCCCCCCCCCCCCCCC(=O)OC XUDJZDNUVZHSKZ-UHFFFAOYSA-N 0.000 description 3
- 238000004445 quantitative analysis Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- TUNFSRHWOTWDNC-UHFFFAOYSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCCC(O)=O TUNFSRHWOTWDNC-UHFFFAOYSA-N 0.000 description 3
- SZHOJFHSIKHZHA-UHFFFAOYSA-N tridecanoic acid Chemical compound CCCCCCCCCCCCC(O)=O SZHOJFHSIKHZHA-UHFFFAOYSA-N 0.000 description 3
- ZQNSHKZQTZSNTB-UHFFFAOYSA-N (3,5-dibromophenyl)methanol Chemical compound OCC1=CC(Br)=CC(Br)=C1 ZQNSHKZQTZSNTB-UHFFFAOYSA-N 0.000 description 2
- IOCYQQQCJYMWDT-UHFFFAOYSA-N (3-ethyl-2-methoxyquinolin-6-yl)-(4-methoxycyclohexyl)methanone Chemical compound C=1C=C2N=C(OC)C(CC)=CC2=CC=1C(=O)C1CCC(OC)CC1 IOCYQQQCJYMWDT-UHFFFAOYSA-N 0.000 description 2
- YUFFSWGQGVEMMI-JLNKQSITSA-N (7Z,10Z,13Z,16Z,19Z)-docosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCCCC(O)=O YUFFSWGQGVEMMI-JLNKQSITSA-N 0.000 description 2
- 235000021294 Docosapentaenoic acid Nutrition 0.000 description 2
- 238000000861 blow drying Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- JFRWATCOFCPIBM-UHFFFAOYSA-N gamma-linolenic acid methyl ester Natural products CCCCCC=CCC=CCC=CCCCCC(=O)OC JFRWATCOFCPIBM-UHFFFAOYSA-N 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- JNSUZRHLHDQGPN-HJWRWDBZSA-N methyl cis-10-heptadecenoate Chemical compound CCCCCC\C=C/CCCCCCCCC(=O)OC JNSUZRHLHDQGPN-HJWRWDBZSA-N 0.000 description 2
- JEDIPLFNJDRCFD-SREVYHEPSA-N methyl cis-10-pentadecenoate Chemical compound CCCC\C=C/CCCCCCCCC(=O)OC JEDIPLFNJDRCFD-SREVYHEPSA-N 0.000 description 2
- AINIZSBLAFHZCP-KHPPLWFESA-N methyl cis-15-tetracosenoate Chemical compound CCCCCCCC\C=C/CCCCCCCCCCCCCC(=O)OC AINIZSBLAFHZCP-KHPPLWFESA-N 0.000 description 2
- JFRWATCOFCPIBM-JPFHKJGASA-N methyl gamma-linolenate Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCC(=O)OC JFRWATCOFCPIBM-JPFHKJGASA-N 0.000 description 2
- VORKGRIRMPBCCZ-UHFFFAOYSA-N methyl tricosanoate Chemical compound CCCCCCCCCCCCCCCCCCCCCCC(=O)OC VORKGRIRMPBCCZ-UHFFFAOYSA-N 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- FPRKGXIOSIUDSE-SYACGTDESA-N (2z,4z,6z,8z)-docosa-2,4,6,8-tetraenoic acid Chemical compound CCCCCCCCCCCCC\C=C/C=C\C=C/C=C\C(O)=O FPRKGXIOSIUDSE-SYACGTDESA-N 0.000 description 1
- ABJFBJGGLJVMAQ-UHFFFAOYSA-N 1,4-dihydroquinoxaline-2,3-dione Chemical compound C1=CC=C2NC(=O)C(=O)NC2=C1 ABJFBJGGLJVMAQ-UHFFFAOYSA-N 0.000 description 1
- CNDLDXJMUUFHDW-UHFFFAOYSA-N CCCCCCCC(=O)OC.CCCCCCCC(=O)OC Chemical compound CCCCCCCC(=O)OC.CCCCCCCC(=O)OC CNDLDXJMUUFHDW-UHFFFAOYSA-N 0.000 description 1
- VHBMMYSRCSWPNB-VDXOJYAPSA-N CCCCCCCCCCCCCC(=O)OC.CCCC\C=C/CCCCCCCC(=O)OC Chemical compound CCCCCCCCCCCCCC(=O)OC.CCCC\C=C/CCCCCCCC(=O)OC VHBMMYSRCSWPNB-VDXOJYAPSA-N 0.000 description 1
- DWWVEZXAPXFPJD-UHFFFAOYSA-N COC(CCCCCCCCCCCCCC)=O.COC(CCCCCCCCCCCCCC)=O Chemical compound COC(CCCCCCCCCCCCCC)=O.COC(CCCCCCCCCCCCCC)=O DWWVEZXAPXFPJD-UHFFFAOYSA-N 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 235000021292 Docosatetraenoic acid Nutrition 0.000 description 1
- 102000012004 Ghrelin Human genes 0.000 description 1
- 101800001586 Ghrelin Proteins 0.000 description 1
- VUVUIDMZOWHIIJ-UHFFFAOYSA-N Methyl-n-nonadecyl-keton Natural products CCCCCCCCCCCCCCCCCCCC(C)=O VUVUIDMZOWHIIJ-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- UPIRHFZFPVEDCF-ZDVGBALWSA-N cis-13,16-Docosadienoic acid methyl ester Chemical compound CCCCC\C=C\C\C=C\CCCCCCCCCCCC(=O)OC UPIRHFZFPVEDCF-ZDVGBALWSA-N 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- FPRKGXIOSIUDSE-UHFFFAOYSA-N docosa-2,4,6,8-tetraenoic acid Chemical compound CCCCCCCCCCCCCC=CC=CC=CC=CC(O)=O FPRKGXIOSIUDSE-UHFFFAOYSA-N 0.000 description 1
- RBKMRGOHCLRTLZ-UHFFFAOYSA-N eicosenoic acid methyl ester Natural products CCCCCCCCC=CCCCCCCCCCC(=O)OC RBKMRGOHCLRTLZ-UHFFFAOYSA-N 0.000 description 1
- ZQPPMHVWECSIRJ-MDZDMXLPSA-N elaidic acid Chemical compound CCCCCCCC\C=C\CCCCCCCC(O)=O ZQPPMHVWECSIRJ-MDZDMXLPSA-N 0.000 description 1
- GNKDKYIHGQKHHM-RJKLHVOGSA-N ghrelin Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)[C@H](CO)NC(=O)CN)COC(=O)CCCCCCC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C1=CC=CC=C1 GNKDKYIHGQKHHM-RJKLHVOGSA-N 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- WCYWZMWISLQXQU-UHFFFAOYSA-N methyl Chemical class [CH3] WCYWZMWISLQXQU-UHFFFAOYSA-N 0.000 description 1
- JFRWATCOFCPIBM-SPOHZTNBSA-N methyl (6e,9e,12e)-octadeca-6,9,12-trienoate Chemical compound CCCCC\C=C\C\C=C\C\C=C\CCCCC(=O)OC JFRWATCOFCPIBM-SPOHZTNBSA-N 0.000 description 1
- YRULPUCCOZGWAE-UHFFFAOYSA-N methyl dodecanoate Chemical compound CCCCCCCCCCCC(=O)OC.CCCCCCCCCCCC(=O)OC YRULPUCCOZGWAE-UHFFFAOYSA-N 0.000 description 1
- YTLQYSWYHSKDDE-UHFFFAOYSA-N methyl tetracos-2-enoate Chemical compound CCCCCCCCCCCCCCCCCCCCCC=CC(=O)OC YTLQYSWYHSKDDE-UHFFFAOYSA-N 0.000 description 1
- 229940105132 myristate Drugs 0.000 description 1
- SECPZKHBENQXJG-FPLPWBNLSA-N palmitoleic acid Chemical compound CCCCCC\C=C/CCCCCCCC(O)=O SECPZKHBENQXJG-FPLPWBNLSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/045—Standards internal
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
-
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Abstract
The invention aims to provide a method for detecting medium-long chain fatty acid in human serum based on gas chromatography-mass spectrometry, which comprises the steps of adding isotope n-nonadecanoic acid methyl ester into a serum sample as an internal standard, then performing a methyl esterification process, extracting the medium-long chain fatty acid in a sample mixture through a liquid-liquid extraction method, separating 40 kinds of medium-long chain fatty acid by using Agilent DB-23 chromatography, and then performing tandem mass spectrometry by using an internal standard curve method. Has the advantages of good selectivity, high sensitivity, short analysis time and the like.
Description
Technical Field
The invention relates to the technical field of metabonomics analysis, in particular to a method for detecting medium-long chain fatty acid in human serum based on gas chromatography-mass spectrometry.
Background
The medium-long chain fatty acid is not only an energy substance, but also a functional substance, influences the growth and development of human beings through mechanisms such as intestinal flora, intestinal morphology, immune function, Ghrelin secretion and the like, and the difference of the influences on the growth performance of human beings is not only related to the content of the medium-long chain fatty acid, but also related to the type of the contained medium-long chain fatty acid, the energy state of the human beings and the health condition of the human beings. In addition, the research of the medium-long chain fatty acid has various physiological effects on preventing and treating chronic diseases and has great significance on maintaining body health.
At present, the GC-MS method is adopted to carry out quantitative analysis on medium-long chain fatty acids, and usually, only less than 37 common medium-long chain fatty acids can be detected, three unsaturated long chain fatty acids, namely docosatetraenoic acid C22:4, docosapentaenoic acid C22:5n-3 and docosapentaenoic acid C22:5n-6, which have important physiological research significance cannot be separated and detected, the development of scientific research is not facilitated, and the achievement is shown, so a new detection method is needed to solve the technical problems.
Disclosure of Invention
The invention aims to provide a method for detecting medium-long chain fatty acid in human serum based on gas chromatography-mass spectrometry, which comprises the steps of adding isotope n-nonadecanoic acid methyl ester into a serum sample as an internal standard, then performing a methyl esterification process, extracting the medium-long chain fatty acid in a sample mixture through a liquid-liquid extraction method, separating 40 kinds of medium-long chain fatty acid by using Agilent DB-23 chromatography, and then performing tandem mass spectrometry by using an internal standard curve method. Has the advantages of good selectivity, high sensitivity, short analysis time and the like.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the method for detecting medium-long chain fatty acid in human serum based on gas chromatography-mass spectrometry comprises the following steps:
s1, preparing an internal standard solution, weighing an isotope internal standard product methyl nonadecanoate, and adding n-hexane to dissolve the isotope internal standard product methyl nonadecanoate to prepare the internal standard solution for later use;
s2, preparing a series of concentration standard substance mixed solution, weighing medium-long chain fatty acid standard substances for gas chromatography-mass spectrometry (GC-MS) after methyl esterification, and preparing a series of concentration gradient standard substance mixed solution by using n-hexane;
step S3, extracting a sample, namely taking a serum sample to be detected, extracting medium-long chain fatty acid in the sample by adopting dichloromethane and methanol, taking a dichloromethane phase, and drying;
step S4, adding an internal standard, redissolving the sample obtained in the step S3 by using normal hexane, adding isotope n-nonadecanoic acid methyl ester (C19:0) as the internal standard, adding potassium hydroxide methanol solution for treatment, adding water, standing for layering, taking supernatant, drying by using nitrogen, and redissolving by using normal hexane to obtain a sample extraction solution;
step S5 gas chromatography-mass spectrometry, wherein the mixed solution of the standard substance with the series concentration gradient obtained in step S2 and the sample extraction solution obtained in step S4 are respectively subjected to gas chromatography, and the mode of the chromatographic temperature gradient is as follows:
heating rate/min | Temperature value of | Residence time min | Run time min | |
Initial temperature | 70 | 2 | 2 | |
Procedure 1 | 20 | 180 | 8 | 15.5 |
Procedure 2 | 4 | 250 | 3 | 36 |
Directly carrying out mass spectrometry on the sample subjected to gas chromatography separation;
and S6, converting the concentration, drawing the mass spectrum data of the standard substance mixed solution with the series of concentration gradients obtained in the step S2 into a standard curve, and converting the mass spectrum data of the sample extraction solution obtained in the step S4 into the corresponding medium-long chain fatty acid concentration through the standard curve.
According to the above scheme, the concentration of the n-nonadecanoic acid methyl ester in the internal standard solution in the step S1 is 1 mg/mL.
According to the scheme, the concentration range of the series concentration standard solution in the step S2 is 1-2500 mg/L.
According to the scheme, the medium-long chain fatty acid standard in the step S2 comprises the following standards: methyl butyrate, methyl hexanoate, methyl octanoate, methyl decanoate, methyl undecanoate, methyl laurate, methyl tridecanoate, methyl myristate, methyl pentadecanoate, methyl palmitate, methyl palmitoleate, methyl heptadecanoate, methyl heptadecenoate, methyl stearate, methyl oleate, methyl elactoate, methyl linoleate, methyl elaidoleate, methyl elanoleate, methyl gama-linolenic acid methyl ester, methyl linolenate, methyl arachidate, methyl eicosenoate, methyl eicosatrienoate, methyl arachidonic acid, methyl eicosapentaenoate, methyl heneicosanoate, methyl docosenoic acid, methyl behenate, methyl erucate, methyl docosedienoate, methyl docosenoic acid, methyl eicosatrinoate, methyl tetracosenoic acid, methyl ester, Methyl tetracosenoic acid, methyl docosatetraenoate, methyl docosapentaenoate and methyl docosapentaenoate.
According to the above scheme, the medium-long chain fatty acid extracted from the sample in step S3 is specifically: adding dichloromethane and methanol into a serum sample to be detected according to a volume ratio of dichloromethane to methanol of 2:1, and uniformly mixing for 1-3min in a vortex manner; oscillating on a shaker for 15-30min at 270rpm at 25 ℃; adding deionized water and anhydrous sodium sulfate, and performing vortex oscillation for 2 min; centrifuging at 2000rpm for 4-6 min; the lower dichloromethane phase was removed and blown dry with nitrogen.
According to the scheme, the detailed steps of adding the internal standard in the step S4 are as follows: adding n-hexane into the sample obtained in the step S3 for redissolving, adding isotope n-nonadecanoic acid methyl ester (C19:0), performing vortex oscillation for 1-3min, adding 0.4mol/L potassium hydroxide methanol solution, performing vortex oscillation for 1-3min, performing oscillation for 20-40min at 37 ℃ by using a shaking table at 180rpm, adding deionized water, performing vortex oscillation for 1-3min, standing for layering, taking the supernatant into a new glass test tube, performing blow-drying by using nitrogen, adding 200 mu L n-hexane, performing vortex oscillation for 2min, standing for layering, and taking the supernatant.
According to the scheme, the medium-long chain fatty acid single-ion detection scanning groups in the mass spectrometry are as follows:
the medium-long chain fatty acid single ion detection scanning parameter conditions in the mass spectrometry are as follows:
and (4) analyzing the standard substance mixed solution with the series of concentration gradients and the sample extraction solution obtained in the step S4 respectively, and deriving and processing mass spectrum data.
The invention has the beneficial effects that: the pretreatment step of the sample can be completed only by simple liquid-liquid extraction, and meanwhile, the important medium-long chain fatty acid substances in serum can be accurately detected by quantifying through an isotope internal standard, the method has high precision and accuracy, can be used for quantitative analysis of clinical serum samples, is simple in experimental operation and short in experimental period, and provides a reliable detection method for health assessment of the medium-long chain fatty acid level in clinic.
Detailed Description
The technical solution of the present invention will be described with reference to the following examples.
The method for detecting medium-long chain fatty acid in human serum based on gas chromatography-mass spectrometry comprises the following steps:
step S1, preparing an internal standard solution, weighing 100mg isotope internal standard product methyl nonadecanoate, adding 1mL of n-hexane to dissolve and prepare 100mg/mL stock solution, taking 10 mu L of stock solution, diluting with n-hexane to a constant volume of 1mL, and preparing 1mg/mL internal standard solution for later use;
s2, preparing a series of concentration standard substance mixed solutions, weighing medium-long chain fatty acid standard substances for gas chromatography-mass spectrometry (GC-MS) after methyl esterification, and preparing a series of concentration gradient standard substance mixed solutions by using n-hexane, wherein the concentration comprises 2500, 1000, 500, 250, 125, 62.5, 25, 10, 5 and 1 mg/L; the medium-long chain fatty acid standard comprises the following standard substances:
methyl butyrate C4:0[623-42-7]2 wt.%;
methyl hexanoate Methyl ester C6:0[106-70-7]2 wt.%;
methyl octanate Methyl octanoate C8:0[111-11-5]2 wt.%;
methyl decanoate Methyl ester C10:0[110-42-9]4 wt.%;
methyl decanoate undecanoate C11:0[1731-86-8]2 wt.%;
methyl dodecanoate Methyl laurate C12:0[111-82-0]4 wt.%;
methyl tricarbacetate tridecanoate C13:0[1731-88-0]2 wt.%;
methyl myrsitate myristate C14:0[124-10-7]2 wt.%;
methyl myristoleate Methyl myristate C14:1, cis-9[56219-06-8]2 wt.%;
methyl pentadecanoate pentadecanoic acid Methyl ester C15:0[7132-64-1]2 wt.%;
methyl cis-10-pentadecenoate pentadecanoic acid Methyl ester C15:1, cis-10[90176-52-6]2 wt.%;
methyl palmitate Methyl ester C16:0[112-39-0]4 wt.%;
methyl palmiteolate palmitoleate C16:1, cis-9[1120-25-8]2 wt.%;
methyl heptadecanoate Methyl ester C17:0[1731-92-6]4 wt.%;
methyl cis-10-heptadecenoate, C17:1, cis-10[75190-82-8]2 wt.%;
methyl stearate C18:0[112-61-8]4 wt.%;
methyl oleate C18:1, cis-9[112-62-9]4 wt.%;
methyl elaidate trans-oleate C18:1T, trans-9[2462-84-2]2 wt.%;
methyl linoleate C18:2, cis-9,12[112-63-0]2 wt.%; (ii) a
Methyl linoleladate Methyl linoleate C18:2TT, trans-9,12[2462-84-2]2 wt.%;
methyl γ -linolenate γ -linolenic acid Methyl ester C18:3, cis-6,9,12[16326-32-2]4 wt.%;
methyl linolenate Methyl ester C18:3, cis-9,12,15[301-00-8]2 wt.%;
methyl arachidate C20:0[1120-28-1]4 wt.%;
cis-11-Eicosenoic acid methyl ester C20:1, cis-11[2390-09-2]2 wt.%;
cis-11,14, 17-eicosarienoic acid methyl ester C20:3, cis-11,14,17[55682-88-7]2 wt.%;
cis-8,11, 14-Eicosaterieno acid methyl ester C20:3, cis-8,11,14[21061-10-9]2 wt.%;
methyl arachidate Methyl arachidonate C20:4, cis-5,8,11,14[2566-89-4]2 wt.%; cis-5,8,11,14,17-Eicosapentaenoic acid methyl ester (EPA-M) C20:5, cis-5,8,11,14,17[2734-47-6]2 wt.%;
methyl hemicosanoate Methyl eicosanoate C21:0[6064-90-0]2 wt.%;
cis-11, 14-Eicosapienoic acid methyl ester C20:2, cis-11,14 [2463-02-7]2 wt.%;
methyl benzoate behenate C22:0[929-77-1]4 wt.%;
methyl erucate Methyl ester C22:1, cis-13[1120-34-9]2 wt.%;
cis-13,16-Docosadienoic acid methyl ester C22:2, cis-13,16 [61012-47-3]2 wt.%;
cis-4,7,10,13,16,19-Docosahexaenoic acid methyl ester (DHA-M) C22:6, cis-4,7,10,13,16,19[301-01-9]2 wt.%;
methyl tricosanoate C23:0[2433-97-8]2 wt.%;
methyl tetracosanoate C24:0[2442-49-1]4 wt.%;
methyl cis-15-tetracosenoate Methyl tetracosenoate C24:1, cis-15[2733-88-2]2 wt.%;
METHYL docosateratate DOCOSATETRAENOATE C22:4[13487-42-8]2 wt.%;
METHYL DOCOSAPENTAENOATE (7C,10C,13C,16C,19C) C22:5n-3 [108698-02-8]2 wt.%;
METHYL DOCOSAPENTAENOATE (4C,7C,10C,13C,16C) C22:5n-6[25182-74-5]2 wt.%;
step S3, extracting a sample, namely putting 100 mu L of a human serum sample to be detected into a 150mL glass test tube, adding 6mL of mixed solution (2:1, v/v) of dichloromethane and methanol into the serum sample to be detected, and uniformly mixing for 2min in a vortex manner; shaking again for 20min at 270rpm and 25 deg.C; then adding 2ml of deionized water and 2g of anhydrous sodium sulfate, and carrying out vortex oscillation for 2 min; centrifuging at 2000rpm for 5 min; taking the dichloromethane phase at the lower layer into a new centrifugal tube, and drying by using nitrogen;
step S4, adding an internal standard, redissolving the sample obtained in the step S3 by using 3mL of normal hexane, adding 200 mL of isotope n-nonadecanoic acid methyl ester (C19:0) with the concentration of 10 mug/mL serving as an internal standard, carrying out vortex oscillation for 2min, adding 3mL of 0.4mol/L potassium hydroxide methanol solution, carrying out vortex oscillation for 2min, oscillating for 30min at the temperature of 37 ℃ by using a shaking table at 180rpm, adding 2mL of deionized water, carrying out vortex oscillation for 2min, standing for layering, taking the supernatant to a new glass test tube, carrying out blow-drying by using nitrogen, adding 200 mug L of normal hexane, carrying out vortex oscillation for 2min, standing for layering, and taking the supernatant. (ii) a
Step S5 gas chromatography-mass spectrometry, wherein the mixed solution of the standard substance with the series concentration gradient obtained in step S2 and the sample extraction solution obtained in step S4 are respectively subjected to gas chromatography, and the mode of the chromatographic temperature gradient is as follows:
heating rate/min | Temperature value of | Residence time min | Run time min | |
Initial temperature | 70 | 2 | 2 | |
Procedure 1 | 20 | 180 | 8 | 15.5 |
Procedure 2 | 4 | 250 | 3 | 36 |
Directly carrying out mass spectrometry on the sample subjected to gas chromatography separation;
the medium-long chain fatty acid single ion detection scanning group in the mass spectrometry is as follows:
the medium-long chain fatty acid single ion detection scanning parameter conditions in the mass spectrometry are as follows:
respectively analyzing the standard substance mixed solution with the series of concentration gradients and the sample extraction solution obtained in the step S4, and deriving mass spectrum data for processing;
and S6, converting the concentration, drawing the mass spectrum data of the standard substance mixed solution with the series of concentration gradients obtained in the step S2 into a standard curve, and converting the mass spectrum data of the sample extraction solution obtained in the step S4 into the corresponding medium-long chain fatty acid concentration through the standard curve.
The test results of this example were further analyzed to verify the feasibility of the method provided by the present invention:
and establishing a standard curve by adopting an isotope internal standard quantitative method and taking the concentration of the medium-long chain fatty acid standard as an x axis and the peak area ratio of the amino acid standard to the internal standard as a y axis. The concentration of medium-long chain fatty acids in serum was calculated from this curve and the results were as follows:
the above embodiments are only used for illustrating but not limiting the technical solutions of the present invention, and although the above embodiments describe the present invention in detail, those skilled in the art should understand that: modifications and equivalents may be made thereto without departing from the spirit and scope of the invention and any modifications and equivalents may fall within the scope of the claims.
Claims (7)
1. The method for detecting medium-long chain fatty acids in human serum based on gas chromatography-mass spectrometry is characterized by comprising the following steps:
s1, preparing an internal standard solution, weighing an isotope internal standard product methyl nonadecanoate, and adding n-hexane to dissolve the isotope internal standard product methyl nonadecanoate to prepare the internal standard solution for later use;
s2, preparing a series of concentration standard substance mixed solution, weighing a medium-long chain fatty acid standard substance for gas chromatography-mass spectrometry after methyl esterification, and preparing a series of concentration gradient standard substance mixed solution by using n-hexane;
step S3, extracting a sample, namely taking a serum sample to be detected, extracting medium-long chain fatty acid in the sample by adopting dichloromethane and methanol, taking a dichloromethane phase, and drying;
step S4, adding an internal standard, redissolving the sample obtained in the step S3 by using normal hexane, adding isotope n-nonadecanoic acid methyl ester as the internal standard, adding a potassium hydroxide methanol solution for treatment, adding water, standing for layering, taking supernatant, drying by blowing with nitrogen, and redissolving by using normal hexane to obtain a sample extraction solution;
step S5 gas chromatography-mass spectrometry, wherein the mixed solution of the standard substance with the series concentration gradient obtained in step S2 and the sample extraction solution obtained in step S4 are respectively subjected to gas chromatography, and the mode of the chromatographic temperature gradient is as follows:
Directly carrying out mass spectrometry on the sample subjected to gas chromatography separation;
and S6, converting the concentration, drawing the mass spectrum data of the standard substance mixed solution with the series of concentration gradients obtained in the step S2 into a standard curve, and converting the mass spectrum data of the sample extraction solution obtained in the step S4 into the corresponding medium-long chain fatty acid concentration through the standard curve.
2. The method for detecting medium-chain fatty acids in human serum based on gas chromatography-mass spectrometry according to claim 1, wherein the concentration of the n-nonadecanoic acid methyl ester in the internal standard solution of step S1 is 1 mg/mL.
3. The method for detecting medium-and long-chain fatty acids in human serum based on gas chromatography-mass spectrometry according to claim 1, wherein the concentration range of the serial concentration standard solution in step S2 is 1-2500 mg/L.
4. The method for detecting medium-long chain fatty acids in human serum based on gas chromatography-mass spectrometry according to claim 1, wherein the medium-long chain fatty acid standard in step S2 comprises the following standards: methyl butyrate, methyl hexanoate, methyl octanoate, methyl decanoate, methyl undecanoate, methyl laurate, methyl tridecanoate, methyl myristate, methyl pentadecanoate, methyl palmitate, methyl palmitoleate, methyl heptadecanoate, methyl heptadecenoate, methyl stearate, methyl oleate, methyl elactoate, methyl linoleate, methyl elaidoleate, methyl elanoleate, methyl gama-linolenic acid methyl ester, methyl linolenate, methyl arachidate, methyl eicosenoate, methyl eicosatrienoate, methyl arachidonic acid, methyl eicosapentaenoate, methyl heneicosanoate, methyl docosenoic acid, methyl behenate, methyl erucate, methyl docosedienoate, methyl docosenoic acid, methyl eicosatrinoate, methyl tetracosenoic acid, methyl ester, Methyl tetracosenoic acid, methyl docosatetraenoate, methyl docosapentaenoate and methyl docosapentaenoate.
5. The method for detecting medium-long chain fatty acids in human serum based on gas chromatography-mass spectrometry according to claim 1, wherein the step of extracting medium-long chain fatty acids from the sample in step S3 specifically comprises: adding dichloromethane and methanol into a serum sample to be detected according to a volume ratio of dichloromethane to methanol of 2:1, and uniformly mixing for 1-3min in a vortex manner; oscillating on a shaker for 15-30min at 270rpm at 25 ℃; adding deionized water and anhydrous sodium sulfate, and performing vortex oscillation for 2 min; centrifuging at 2000rpm for 4-6 min; the lower dichloromethane phase was removed and blown dry with nitrogen.
6. The method for detecting medium-long chain fatty acids in human serum based on gas chromatography-mass spectrometry as claimed in claim 1, wherein the step S4 is detailed by adding an internal standard: adding n-hexane into the sample obtained in the step S3 for redissolving, adding isotope n-nonadecanoic acid methyl ester, performing vortex oscillation for 1-3min, adding 0.4mol/L potassium hydroxide methanol solution, performing vortex oscillation for 1-3min, performing oscillation for 20-40min at 37 ℃ by using a shaking table at 180rpm, adding deionized water, performing vortex oscillation for 1-3min, standing for layering, taking the supernatant into a new glass test tube, drying by using nitrogen, adding 200 mu L n-hexane, performing vortex oscillation for 2min, standing for layering, and taking the supernatant.
7. The method for detecting medium-long chain fatty acids in human serum based on gas chromatography-mass spectrometry as claimed in claim 1, wherein the medium-long chain fatty acid single ion detection scans in mass spectrometry are grouped as follows:
the medium-long chain fatty acid single ion detection scanning parameter conditions in the mass spectrometry are as follows:
and (4) analyzing the standard substance mixed solution with the series of concentration gradients and the sample extraction solution obtained in the step S4 respectively, and deriving and processing mass spectrum data.
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