CN113655223A - Multiplex amino acid quantitative method and kit development - Google Patents
Multiplex amino acid quantitative method and kit development Download PDFInfo
- Publication number
- CN113655223A CN113655223A CN202110710806.XA CN202110710806A CN113655223A CN 113655223 A CN113655223 A CN 113655223A CN 202110710806 A CN202110710806 A CN 202110710806A CN 113655223 A CN113655223 A CN 113655223A
- Authority
- CN
- China
- Prior art keywords
- amino acid
- kit
- acid
- amino acids
- standard amino
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 150000001413 amino acids Chemical class 0.000 title claims abstract description 55
- 238000004445 quantitative analysis Methods 0.000 title claims abstract description 9
- 238000011161 development Methods 0.000 title description 2
- 238000000034 method Methods 0.000 claims abstract description 25
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 17
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000006243 chemical reaction Methods 0.000 claims abstract description 5
- 150000002500 ions Chemical class 0.000 claims abstract description 5
- 238000002552 multiple reaction monitoring Methods 0.000 claims abstract 5
- 238000012544 monitoring process Methods 0.000 claims abstract 2
- 235000001014 amino acid Nutrition 0.000 claims description 49
- 229940024606 amino acid Drugs 0.000 claims description 49
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 24
- 238000002372 labelling Methods 0.000 claims description 20
- 239000003153 chemical reaction reagent Substances 0.000 claims description 19
- 239000000047 product Substances 0.000 claims description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 16
- 239000006228 supernatant Substances 0.000 claims description 15
- 238000003908 quality control method Methods 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 12
- 235000018102 proteins Nutrition 0.000 claims description 10
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 102000004169 proteins and genes Human genes 0.000 claims description 10
- 210000002966 serum Anatomy 0.000 claims description 10
- 238000001819 mass spectrum Methods 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 8
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 7
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 7
- 238000001228 spectrum Methods 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 claims description 4
- QCHPKSFMDHPSNR-UHFFFAOYSA-N 3-aminoisobutyric acid Chemical compound NCC(C)C(O)=O QCHPKSFMDHPSNR-UHFFFAOYSA-N 0.000 claims description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 4
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 claims description 4
- JDHILDINMRGULE-LURJTMIESA-N N(pros)-methyl-L-histidine Chemical compound CN1C=NC=C1C[C@H](N)C(O)=O JDHILDINMRGULE-LURJTMIESA-N 0.000 claims description 4
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 claims description 4
- 238000004949 mass spectrometry Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- FSYKKLYZXJSNPZ-UHFFFAOYSA-N sarcosine Chemical compound C[NH2+]CC([O-])=O FSYKKLYZXJSNPZ-UHFFFAOYSA-N 0.000 claims description 4
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 claims description 4
- 210000004369 blood Anatomy 0.000 claims description 3
- 239000008280 blood Substances 0.000 claims description 3
- 210000000601 blood cell Anatomy 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 claims description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims description 2
- LUTLAXLNPLZCOF-UHFFFAOYSA-N 1-Methylhistidine Natural products OC(=O)C(N)(C)CC1=NC=CN1 LUTLAXLNPLZCOF-UHFFFAOYSA-N 0.000 claims description 2
- BRMWTNUJHUMWMS-UHFFFAOYSA-N 3-Methylhistidine Natural products CN1C=NC(CC(N)C(O)=O)=C1 BRMWTNUJHUMWMS-UHFFFAOYSA-N 0.000 claims description 2
- 108010085443 Anserine Proteins 0.000 claims description 2
- 239000004475 Arginine Substances 0.000 claims description 2
- KDZOASGQNOPSCU-WDSKDSINSA-N Argininosuccinic acid Chemical compound OC(=O)[C@@H](N)CCC\N=C(/N)N[C@H](C(O)=O)CC(O)=O KDZOASGQNOPSCU-WDSKDSINSA-N 0.000 claims description 2
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 claims description 2
- QRYRORQUOLYVBU-VBKZILBWSA-N Carnosic acid Natural products CC([C@@H]1CC2)(C)CCC[C@]1(C(O)=O)C1=C2C=C(C(C)C)C(O)=C1O QRYRORQUOLYVBU-VBKZILBWSA-N 0.000 claims description 2
- 108010087806 Carnosine Proteins 0.000 claims description 2
- YPWSLBHSMIKTPR-UHFFFAOYSA-N Cystathionine Natural products OC(=O)C(N)CCSSCC(N)C(O)=O YPWSLBHSMIKTPR-UHFFFAOYSA-N 0.000 claims description 2
- ILRYLPWNYFXEMH-UHFFFAOYSA-N D-cystathionine Natural products OC(=O)C(N)CCSCC(N)C(O)=O ILRYLPWNYFXEMH-UHFFFAOYSA-N 0.000 claims description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 2
- 239000004471 Glycine Substances 0.000 claims description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 2
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 claims description 2
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 2
- SLRNWACWRVGMKD-UHFFFAOYSA-N L-anserine Natural products CN1C=NC(CC(NC(=O)CCN)C(O)=O)=C1 SLRNWACWRVGMKD-UHFFFAOYSA-N 0.000 claims description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 claims description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 2
- ILRYLPWNYFXEMH-WHFBIAKZSA-N L-cystathionine Chemical compound [O-]C(=O)[C@@H]([NH3+])CCSC[C@H]([NH3+])C([O-])=O ILRYLPWNYFXEMH-WHFBIAKZSA-N 0.000 claims description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 2
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims description 2
- FFFHZYDWPBMWHY-VKHMYHEASA-N L-homocysteine Chemical compound OC(=O)[C@@H](N)CCS FFFHZYDWPBMWHY-VKHMYHEASA-N 0.000 claims description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 2
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 2
- LRQKBLKVPFOOQJ-YFKPBYRVSA-N L-norleucine Chemical compound CCCC[C@H]([NH3+])C([O-])=O LRQKBLKVPFOOQJ-YFKPBYRVSA-N 0.000 claims description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims description 2
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 claims description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 2
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 2
- 239000004472 Lysine Substances 0.000 claims description 2
- CQOVPNPJLQNMDC-UHFFFAOYSA-N N-beta-alanyl-L-histidine Natural products NCCC(=O)NC(C(O)=O)CC1=CN=CN1 CQOVPNPJLQNMDC-UHFFFAOYSA-N 0.000 claims description 2
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 claims description 2
- SUHOOTKUPISOBE-UHFFFAOYSA-N O-phosphoethanolamine Chemical compound NCCOP(O)(O)=O SUHOOTKUPISOBE-UHFFFAOYSA-N 0.000 claims description 2
- BZQFBWGGLXLEPQ-UHFFFAOYSA-N O-phosphoryl-L-serine Natural products OC(=O)C(N)COP(O)(O)=O BZQFBWGGLXLEPQ-UHFFFAOYSA-N 0.000 claims description 2
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 claims description 2
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 claims description 2
- 241000210053 Potentilla elegans Species 0.000 claims description 2
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 2
- 108010077895 Sarcosine Proteins 0.000 claims description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 2
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims description 2
- 239000004473 Threonine Substances 0.000 claims description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims description 2
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims description 2
- 235000004279 alanine Nutrition 0.000 claims description 2
- QWCKQJZIFLGMSD-UHFFFAOYSA-N alpha-aminobutyric acid Chemical compound CCC(N)C(O)=O QWCKQJZIFLGMSD-UHFFFAOYSA-N 0.000 claims description 2
- MYYIAHXIVFADCU-QMMMGPOBSA-N anserine Chemical compound CN1C=NC=C1C[C@H](NC(=O)CC[NH3+])C([O-])=O MYYIAHXIVFADCU-QMMMGPOBSA-N 0.000 claims description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 2
- 235000009582 asparagine Nutrition 0.000 claims description 2
- 229960001230 asparagine Drugs 0.000 claims description 2
- 235000003704 aspartic acid Nutrition 0.000 claims description 2
- 229960005261 aspartic acid Drugs 0.000 claims description 2
- 229940000635 beta-alanine Drugs 0.000 claims description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 2
- 229940044199 carnosine Drugs 0.000 claims description 2
- CQOVPNPJLQNMDC-ZETCQYMHSA-N carnosine Chemical compound [NH3+]CCC(=O)N[C@H](C([O-])=O)CC1=CNC=N1 CQOVPNPJLQNMDC-ZETCQYMHSA-N 0.000 claims description 2
- 229960002173 citrulline Drugs 0.000 claims description 2
- 235000013477 citrulline Nutrition 0.000 claims description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 2
- 235000018417 cysteine Nutrition 0.000 claims description 2
- 229960002433 cysteine Drugs 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- 229950006137 dexfosfoserine Drugs 0.000 claims description 2
- YSMODUONRAFBET-UHNVWZDZSA-N erythro-5-hydroxy-L-lysine Chemical compound NC[C@H](O)CC[C@H](N)C(O)=O YSMODUONRAFBET-UHNVWZDZSA-N 0.000 claims description 2
- 229960003692 gamma aminobutyric acid Drugs 0.000 claims description 2
- 235000013922 glutamic acid Nutrition 0.000 claims description 2
- 239000004220 glutamic acid Substances 0.000 claims description 2
- 229960002989 glutamic acid Drugs 0.000 claims description 2
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims description 2
- 229960002743 glutamine Drugs 0.000 claims description 2
- 235000004554 glutamine Nutrition 0.000 claims description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 2
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims description 2
- 229960000310 isoleucine Drugs 0.000 claims description 2
- 238000001948 isotopic labelling Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 claims description 2
- 229930182817 methionine Natural products 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 229960003104 ornithine Drugs 0.000 claims description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 2
- BZQFBWGGLXLEPQ-REOHCLBHSA-N phosphoserine Chemical compound OC(=O)[C@@H](N)COP(O)(O)=O BZQFBWGGLXLEPQ-REOHCLBHSA-N 0.000 claims description 2
- 230000001376 precipitating effect Effects 0.000 claims description 2
- 238000012545 processing Methods 0.000 claims description 2
- 229960002429 proline Drugs 0.000 claims description 2
- 150000003148 prolines Chemical class 0.000 claims description 2
- 229940043230 sarcosine Drugs 0.000 claims description 2
- 229960003080 taurine Drugs 0.000 claims description 2
- 239000004474 valine Substances 0.000 claims description 2
- 238000002474 experimental method Methods 0.000 claims 2
- 239000003550 marker Substances 0.000 claims 2
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 claims 1
- 229910000013 Ammonium bicarbonate Inorganic materials 0.000 claims 1
- 235000012538 ammonium bicarbonate Nutrition 0.000 claims 1
- 239000001099 ammonium carbonate Substances 0.000 claims 1
- WSFSSNUMVMOOMR-DICFDUPASA-N dideuteriomethanone Chemical compound [2H]C([2H])=O WSFSSNUMVMOOMR-DICFDUPASA-N 0.000 claims 1
- 239000000463 material Substances 0.000 claims 1
- 239000008055 phosphate buffer solution Substances 0.000 claims 1
- 108090000765 processed proteins & peptides Proteins 0.000 claims 1
- 229910000033 sodium borohydride Inorganic materials 0.000 claims 1
- 239000012279 sodium borohydride Substances 0.000 claims 1
- MNWBNISUBARLIT-UHFFFAOYSA-N sodium cyanide Chemical compound [Na+].N#[C-] MNWBNISUBARLIT-UHFFFAOYSA-N 0.000 claims 1
- -1 sodium cyanoborohydrides Chemical class 0.000 claims 1
- 230000003595 spectral effect Effects 0.000 claims 1
- 238000001196 time-of-flight mass spectrum Methods 0.000 claims 1
- 210000003462 vein Anatomy 0.000 claims 1
- 230000000155 isotopic effect Effects 0.000 abstract description 3
- 238000011002 quantification Methods 0.000 abstract description 3
- 230000001419 dependent effect Effects 0.000 abstract 1
- 230000004907 flux Effects 0.000 abstract 1
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 10
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 5
- 235000019253 formic acid Nutrition 0.000 description 5
- 239000012224 working solution Substances 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 3
- 238000000861 blow drying Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 238000007086 side reaction Methods 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O ammonium group Chemical group [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000012488 skeletal system development Effects 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6806—Determination of free amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Hematology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Bioinformatics & Computational Biology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention relates to a novel Multiple Amino Acid Quantification (MAAQ) method and a related kit. The technical scheme of the invention is as follows: the amino acids are chemically labeled by organic combination of formaldehyde and sodium cyanoborohydride and their respective isotopic forms, so that the amino acids are provided with different mass labels, and quantitative analysis of the amino acids in multiple samples is realized by selective ion Scanning (SIM), Multiple Reaction Monitoring (MRM), Parallel Reaction Monitoring (PRM) or non-data dependent acquisition (DIA). The method and the kit have the advantages of high quantitative flux, low cost, high accuracy and the like, and the kit can be used for greatly facilitating the quantification of the amino acid.
Description
Technical Field
The invention designs an amino acid quantitative method based on mass spectrum technology, which can realize large-scale, high-accuracy and high-precision quantification of amino acid samples and has the advantages of low price, simple method marking, high marking efficiency, no side reaction and the like. The method can be used for clinical diagnosis and quantitative analysis of amino acids contained in food samples.
Background
Amino acids are important components of life and are constituent units of proteins. There are more than 40 kinds of important amino acids in human body, including 20 kinds of protein amino acids and some amino acids with important functions, which are collectively called "full spectrum amino acids". Specifically, the compound comprises arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, gamma-aminobutyric acid, glycine, serine, taurine, tyrosine, alpha-aminocaproic acid, asparagine, aspartic acid, citrulline, glutamic acid, glutamine, ornithine, cysteine, cystathionine, homocysteine, alpha-aminon-butyric acid, alanine, anserine, beta-alanine, beta-aminoisobutyric acid, carnosine, ethanolamine, delta-hydroxylysine, hydroxylated proline, 1-methylhistidine, 3-methylhistidine, phosphoethanolamine, phosphoserine, proline, sarcosine, argininosuccinic acid and hydroxylated citrulline. The full spectrum of amino acids is related to many diseases, such as metabolic diseases, tumors, immunity, cardiovascular and cerebrovascular diseases, nervous system diseases and the like, and is also related to skeletal development, hormone secretion and the like of children. In addition, the content of amino acids is also required to be tested in food testing to determine the efficacy of the food.
The existing quantitative method mainly combines a derivation method with a liquid chromatography, a gas chromatography-mass spectrometry and a liquid chromatography-mass spectrometry combined method. The liquid chromatography-mass spectrometry combined method can simultaneously realize qualitative and quantitative determination due to simple sample treatment, and is widely applied. The application of the liquid chromatography-mass spectrometry method is based on a stable isotope labeling addition method, and stable isotopes can be introduced by adding amino acids containing 13C, 15N or 2H elements in the sample pretreatment process and carrying out chemical labeling by using reagents containing the elements. However, the detection cost and the throughput of the method are low, so that the clinical application of the method is limited.
Disclosure of Invention
Therefore, a novel labeling method is designed, organic combination is carried out by using formaldehyde, sodium cyanoborohydride and isotope forms thereof, as shown in table 1, the simultaneous quantitative analysis of the highest eight samples is realized, and the analysis cost of a single sample is lower than 1/10 of the existing analysis method.
TABLE 1 summary of the combined labeling of formaldehyde and sodium cyanoborohydride containing different isotopic labels
In order to achieve the purpose, the invention adopts the technical scheme that:
1. preparation of internal standard amino acid: any one of 8 labeling combinations of labeled standard amino acids is used as an internal standard amino acid, and Dim36 labeled standard amino acids are selected in the kit.
2. Preparing a quality control product: labeling standard amino acid with labeling reagent different from the internal standard amino acid, and adding into the serum dry powder. The kit selectively uses Dim28 to mark standard amino acid, and adds internal standard amino acid.
3. Preparing a calibration product: dim28, Dim30H, Dim32H, Dim34H and Dim36 are selected to mark standard amino acids respectively and are mixed together in a certain concentration gradient and added into serum dry powder.
4. Clinical sample amino acid extraction: blood was taken intravenously, blood cells were removed by centrifugation, and the supernatant was collected. Adding acetonitrile or methanol into the supernatant to precipitate protein, centrifuging to obtain supernatant, blow drying, redissolving, and adding a labeling reagent to make free amino acid react with a chemical label. The labeling reagent cannot be identical to the internal standard amino acid.
5. And re-dissolving the quality control product and the calibrator. Adding acetonitrile or methanol, precipitating protein, and collecting supernatant.
6. Mass spectrometry analysis: and (4) drying the supernatants of the quality control product and the calibrator by using nitrogen, and redissolving by using a mobile phase A liquid of a chromatogram. And (3) respectively carrying out mass spectrum analysis on the clinical sample, the quality control product and the calibrator.
The invention has the following advantages:
1. the sample pretreatment is simple, and the sample pretreatment process of the method is the same as the conventional sample pretreatment process.
2. The marking method is simple, efficient, rapid, high in reaction effect and free of side reaction, and can finish marking of the sample within 5 minutes, and the marking efficiency is 100%.
3. The labeling method has low cost of labeling samples, and the labeling price of 100 mu mol of amino acid samples is much lower than the price of an iTRAQ reagent and the price of directly using stable isotope labeled amino acid.
Description of the drawings:
1. FIG. 1 is a schematic representation of the products and mass gain of amino acids labeled with formaldehyde and sodium cyanoborohydride of different isotopic composition.
Detailed Description
1. Preparation of internal standard amino acid: mixing 10 μ L4%13CD2O and 10. mu.L of 0.6mol/L NaCNBD3The labeling reagent of (2) was added to 100. mu.L of a 0.01M aqueous tyrosine solution, reacted at room temperature for 15min, followed by addition of 10. mu.L of 1M NH4HCO3The solution was reacted for 10min to remove excess labeling reagent. The solution after the above reaction was used as an internal standard stock solution, and the working solution was diluted 1000 times with 0.1% formic acid to be used as a working solution.
2. Preparing a quality control product: 10 μ L of 4% CH2O and 10. mu.L of 0.6mol/L NaCNBH3The labeling reagent of (2) was added to 100. mu.L of a 0.01M aqueous tyrosine solution, reacted at room temperature for 15min, followed by addition of 10. mu.L of 1M NH4HCO3The solution was reacted for 10min to remove excess labeling reagent, the working solution was diluted 500-fold with 0.1% formic acid, and 10. mu.L of the solution was added to the serum dry powder. Adding 10 μ L of internal standard working solution into the serum dry powder, dissolving the serum dry powder with 0.1mL of 0.1% formic acid, adding 400 μ L of acetonitrile to precipitate protein, centrifuging at 15000g, removing protein, collecting supernatant, blow-drying with nitrogen, and redissolving with 100 μ L of 0.1% formic acid for later use.
3. Preparing a calibration product: in the same manner, the tyrosine solutions were labeled with Dim28, Dim30H, Dim32H, Dim34H and Dim36 reagents, respectively. And the ratio of Dim 28: dim 30H: dim 32H: dim 34H: mixing Dim36 ═ 2:10:50:250:500, adding into 100 μ L serum solution, adding 400 μ L acetonitrile to precipitate protein, centrifuging at 15000g, removing protein, collecting supernatant, blowing with nitrogen, and redissolving with 100 μ L0.1% formic acid for use.
4. Clinical sample amino acid extraction: blood was collected intravenously, and the supernatant was collected by removing blood cells by centrifugation. Collecting 100 μ L supernatant, adding 400 μ L acetonitrile to precipitate protein15000g for 10min, collecting the supernatant, adding 10 μ L of 4% CH2O and 10. mu.L of 0.6mol/L NaCNBH3Reaction at room temperature for 15min, followed by addition of 10. mu.L of 1M NH4HCO3And (3) reacting the solution for 10min to remove redundant marking reagents, adding 10 mu L of internal standard working solution, desalting, drying, and redissolving by using 100 mu L of mobile phase A solution for later use.
5. Mass spectrometry analysis: and (3) respectively carrying out mass spectrum analysis on the clinical sample, the quality control product and the calibrator. Liquid chromatography conditions: ACQUITY UPLC I-Class with column oven (CH-A), sample size 1. mu.L, column temperature: 40 ℃; column ACQUITY UPLCR BEH C18,1.7 μm, 2.1X 100mm, flow rate 0.2mL/min, mobile phase A: water + 0.1% FA; mobile phase B: meoh + 0.1% FA +2mM ammonium form. Gradiometer:
Time | %B | Curve |
0 | 2 | 6 |
2 | 2 | 6 |
3.5 | 10 | 6 |
4 | 80 | 6 |
4.3 | 2 | 6 |
4.5 | 2 | 6 |
mass spectrum conditions: xevo TQD; acquisition mode MRM, parent ion 210> 164; 212> 166; 214> 168; 216> 170; 218> 172. Polarity ESI +; capillary voltage 0.5 kV; the ion source temperature is 150 ℃; desolventizing gas temperature: 350 ℃; cone voltage 16V, impact energy 16V.
Data processing: MassLynx software version 4.1 with TargetLynx application management software
Evaluation of the method:
1. external standard curve: the method utilizes the addition of the amino acid marked by five isotopes into the same sample and the mixing of the amino acid with different concentration gradients, thereby overcoming the defects of low linear correlation coefficient of a quantitative curve and large consumption of machine time caused by multiple sample injection. Tyrosine was labeled in this example with Dim28, Dim30H, Dim32H, Dim34H and Dim36, labeled as Dim 28: dim 30H: dim 32H: dim 34H: dim36 was mixed at 2:10:50:250:500 and quantified by fragment ion peak area.
2. Quantifying quality control products: the content of the amino acid marked by Dim28 in the quality control product is 2 times of that of the amino acid marked by the internal standard Dim 36. The actual measurement result is 1.98 times, which shows extremely high quantitative accuracy.
Use of the MAAQ method in clinical sample analysis
Serum of healthy persons selected from 10 physical examination samples was analyzed by the MAAQ method, and the tyrosine content was 57.4 + -7.9. mu. mol/L.
In the description of the present invention, it should be noted that the terms "upper", "lower", "inner", "outer", "top/bottom", and the like indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience of description and simplification of description, but do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus should not be construed as limiting the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "disposed," "sleeved/connected," "connected," and the like are to be construed broadly, e.g., "connected," which may be fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (7)
1. A multiple amino acid quantitative Method (MAAQ) is characterized in that formaldehyde, sodium cyanoborohydride and respective isotope forms thereof are organically combined to carry out chemical labeling on amino acid, so that the amino acid to be detected is provided with different mass labels, the peak intensity of a primary spectrum or a secondary spectrum of a mass spectrum is utilized to carry out quantitative analysis, the flow in an experiment is immobilized, and required reagents and consumables are assembled into a kit to facilitate popularization and use of the method.
2. The method of claim 1, wherein: the organic combination of formaldehyde and sodium cyanoborohydride and the isotope form thereof is carried out to label the peptide segments, namely four typesFormaldehyde (CH)2O,13CH2O,CD2O and13CD2o) and two sodium cyanoborohydrides (NaCNBH)3And NaCNBD3) The combination was performed to form 8 labels, resulting in different quality tags, as shown in the following table.
3. The method of claim 1, the amino acids being characterized by: the full-spectrum amino acids in organisms specifically include: arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, gamma-aminobutyric acid, glycine, serine, taurine, tyrosine, alpha-aminocaproic acid, asparagine, aspartic acid, citrulline, glutamic acid, glutamine, ornithine, cysteine, cystathionine, homocysteine, alpha-aminon-butyric acid, alanine, anserine, beta-alanine, beta-aminoisobutyric acid, carnosine, ethanolamine, delta-hydroxylysine, hydroxylated proline, 1-methylhistidine, 3-methylhistidine, phosphoethanolamine, phosphoserine, proline, sarcosine, argininosuccinic acid, hydroxylated citrulline.
4. The method of claim 1, wherein: the mass spectrum acquisition method mainly comprises a triple quadrupole mass spectrum, a quadrupole-time-of-flight mass spectrum and a quadrupole-electrostatic orbitrap mass spectrum, and comprises Multiple Reaction Monitoring (MRM), Parallel Reaction Monitoring (PRM), selective ion detection (SIM) and full scan (full scan).
5. The kit of claim 1, wherein: the kit comprises reagents required by experiments and required consumables, wherein the reagents comprise a calibrator, a quality control product, serum dry powder, an isotope labeling reagent, four kinds of formaldehyde and two kinds of cyano sodium borohydride, 1mol/L ammonium bicarbonate solution (pH 7.8), 50mmol/L phosphate buffer solution, acetonitrile, methanol and a chromatographic mobile phase; the consumables included 1.7mL centrifuge tubes.
6. Kit according to claims 1 and 5, comprising the following processing steps:
1) preparation of internal standard amino acid: any one of 8 marker combinations is used as an internal standard amino acid, and the kit selects Dim36 marker standard amino acids;
2) preparing a quality control product: marking standard amino acid by using a marking reagent different from the internal standard amino acid, adding the standard amino acid into the serum dry powder, and selectively marking the standard amino acid by using Dim 28; adding internal standard amino acid;
3) preparing a calibration product: selecting Dim28, Dim30H, Dim32H, Dim34H and Dim36 as markers of standard amino acids, mixing the materials together at a ratio of 1:5:10:50:100, and adding the mixture into serum dry powder;
4) clinical sample amino acid extraction: taking blood from vein, centrifuging to remove blood cells, and collecting supernatant; adding acetonitrile or methanol into the supernatant to precipitate protein, taking the supernatant, drying, redissolving, and adding a labeling reagent to enable free amino acid to react to form a chemical label; and adding the internal standard substance of the step 1; the kit uses Dim28, Dim30L, Dim30H, Dim32L, Dim32H, Dim34L and Dim34H to mark clinical samples;
5) re-dissolving the quality control product and the calibrator; adding the quality control product and the calibrator obtained in the steps 2 and 3 into acetonitrile or methanol, precipitating protein, and taking supernatant;
6) mass spectrometry analysis: drying the supernatants of the quality control product and the calibrator obtained in the step 5 by using nitrogen, and redissolving by using mobile phase A liquid of a chromatogram; and (4) respectively carrying out mass spectrum analysis on the clinical sample, the quality control product and the calibrator obtained in the step (4) and the step (5).
7. The kit according to claim 6, the labeling method of step 4, having the following characteristics: when using the SIM or MRM mode of the serial quadrupole of claim 4, the labeling reagent is selected from Dim28, Dim30H, Dim32H, Dim 34H; when selecting PRM, SIM or full scan of electrostatic orbitrap mass spectrometry, the labeling reagents were selected from Dim28, Dim30L, Dim30H, Dim32L, Dim32H, Dim34L, Dim34H, while the mass spectral resolution was greater than 60000 when m/z is 200.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110710806.XA CN113655223A (en) | 2021-06-25 | 2021-06-25 | Multiplex amino acid quantitative method and kit development |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110710806.XA CN113655223A (en) | 2021-06-25 | 2021-06-25 | Multiplex amino acid quantitative method and kit development |
Publications (1)
Publication Number | Publication Date |
---|---|
CN113655223A true CN113655223A (en) | 2021-11-16 |
Family
ID=78488993
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110710806.XA Pending CN113655223A (en) | 2021-06-25 | 2021-06-25 | Multiplex amino acid quantitative method and kit development |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113655223A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114740121A (en) * | 2022-04-24 | 2022-07-12 | 杭州广科安德生物科技有限公司 | Double-amino-acid stable isotope labeling method based on primary mass spectrometry and application |
-
2021
- 2021-06-25 CN CN202110710806.XA patent/CN113655223A/en active Pending
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114740121A (en) * | 2022-04-24 | 2022-07-12 | 杭州广科安德生物科技有限公司 | Double-amino-acid stable isotope labeling method based on primary mass spectrometry and application |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7399214B2 (en) | Amino acid analysis in body fluids by liquid chromatography mass spectrometry | |
Warren | Organic N molecules in the soil solution: what is known, what is unknown and the path forwards | |
Sidoli et al. | One minute analysis of 200 histone posttranslational modifications by direct injection mass spectrometry | |
CA2889411C (en) | Neutron encoded mass tags for analyte quantification | |
WO2005116629A1 (en) | Method and apparatus for analyzing aminofunctional compound | |
CN103616454A (en) | Method and kit for quantitatively detecting human beta-casein content | |
CN113063871B (en) | Metabolic small molecule detection method and detection system | |
CN114236025A (en) | Liquid phase mass spectrum method for simultaneously determining 43 amino acids without using ion pair reagent and non-derivatization | |
US9012835B2 (en) | Methods for simultaneous quantification of thyroid hormones and metabolites thereof by mass spectrometry | |
CN110678756B (en) | Method for absolute quantification of low abundance polypeptides using mass spectrometry | |
CN113655223A (en) | Multiplex amino acid quantitative method and kit development | |
CN113607854A (en) | Method and detection kit for simultaneously detecting multiple vitamins | |
US20040096876A1 (en) | Quantitative analysis via isotopically differentiated derivatization | |
CN116223693B (en) | Method for measuring folic acid and metabolite thereof in erythrocytes by high performance liquid chromatography tandem mass spectrometry | |
CN105092733B (en) | The reduction method and apparatus of fixedness buffer salt content in LC MS testers | |
CN116297993A (en) | Kit for determining folic acid and metabolite thereof in erythrocytes by high performance liquid chromatography tandem mass spectrometry | |
Sakaguchi et al. | Quantification of peptides using N‐terminal isotope coding and C‐terminal derivatization for sensitive analysis by micro liquid chromatography‐tandem mass spectrometry | |
CN112526048B (en) | Method for rapidly detecting trace residues of hypertensive drugs in environmental sediment | |
EP3911959B1 (en) | High speed sample workflow for lc-ms based hba1c measurement | |
CN110498838B (en) | Characteristic peptide segment for detecting FPGS (planar-repeats-GS) and GGH (GGH) protein expression level and application thereof | |
US9057698B2 (en) | Methods of chemoselective derivation of multiple classes of metabolites | |
CN114577916A (en) | Analysis method of stable isotope labeling metabolic flow based on chip nanoliter electrospray mass spectrum | |
Richards et al. | Quantitative analysis with modern bioanalytical mass spectrometry and stable isotope labeling | |
CN114563504B (en) | Method and kit for determining content of free aldosterone in blood plasma | |
CN104991028B (en) | The reduction method of fixedness buffer salt content in LC MS testers |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |