QUANTITATIVE DETECTION METHOD OF MULTIPLE METABOLITES IN BIOLOGICAL SAMPLE AND METABOLIC CHIP

Abstract

The present invention discloses a quantitative detection method of multiple metabolic components in a biological sample and a metabolic chip used in the method. The detection method includes performing derivatization treatment on the biological sample and then detecting the derivatized biological sample by liquid chromatography-mass spectrometry. The metabolic chip of the present invention includes a chip carrier microtiter plate and related reagents, and quantitative detection of multiple metabolic components of different magnitudes such as amino acid, phenol, phenyl or benzyl derivative, indole, organic acid, fatty acid, sugar, and bile acid in the biological sample on the same microtiter plate can be achieved.

Claims

1. A quantitative detection method of multiple metabolic components in a biological sample, comprising performing derivatization treatment on the biological sample and then detecting the derivatized biological sample by liquid chromatography-mass spectrometry, wherein during derivatization treatment, 3-nitrophenylhydrazine is used as a derivatization reagent, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide is used as a derivatization reaction catalyst; the biological sample is selected from urine, blood, cerebrospinal fluid, tissue, cells, saliva and fecal samples of a mammal; the multiple metabolic components in the biological sample detected by the quantitative detection method are selected from amino acid, phenol, phenyl or benzyl derivative, indole, organic acid, fatty acid, sugar, and bile acid and have different magnitudes in content; the number of the multiple metabolic components in a biological sample detected by the quantitative detection method is 50 or more; the liquid chromatography adopts a UPLC BEH C18 chromatographic column, the mobile phase includes: A: water containing 0.1% formic acid, B: acetonitrile containing 0.1% formic acid and isopropanol at a ratio of 1-2:1; and the gradient elution conditions: 0-1 min, 5% B; 1-5 min, 5-30% B; 5-9 min, 30-50% B; 9-12 min, 50-75% B; 12-15 min, 75-95% B; 15-16 min, 95-100% B; 16-18 min, 100% B; and 18-20 min, 5% B.

2. The detection method according to claim 1, wherein the number of the multiple metabolic components in a biological sample detected by the quantitative detection method is 100 or more.

3. The detection method according to claim 1, wherein the number of the multiple metabolic components in a biological sample detected by the quantitative detection method is 200 or more.

4. The detection method according to claim 1, wherein the number of the multiple metabolic components in a biological sample detected by the quantitative detection method is 600 or more.

5. The detection method according to claim 1, wherein the number of the multiple metabolic components in a biological sample detected by the quantitative detection method is 1000 or more.

6. The detection method according to claim 1, wherein scan parameters of the mass spectrometry are set as follows: S-lens RF level, 50; mass range, 100 to 1200 m/z; full MS resolution, 70,000; MS/MS resolution, 17,500; and NCE/stepped NCE is set at 10, 20, and 40 eV.

7. The detection method according to claim 1, wherein ESI ion source parameters of the mass spectrometry are set as follows: spray voltage, 3800 V; sheath gas flow rate, 40 for ESI+ and 5 for ESI; capillary temperature, 320 C.; probe heater temperature, 350 C.; aux gas flow rate, 10.

8. The detection method according to claim 1, wherein the liquid chromatography-mass spectrometry is acquired using a Vanquish Flex UHPLC system coupled to a Q Extractive Focus Orbitrap mass spectrometer equipped with a heated electrospray ionization source.

9. The detection method according to claim 1, wherein the mobile phase includes: A: water containing 0.1% formic acid, B: acetonitrile containing 0.1% formic acid and isopropanol at a ratio of 7:3.

10. The detection method according to claim 1, comprising the following steps: a) collecting a biological sample; b) extracting the biological sample with a mixed solvent of methanol, chloroform, and water, performing centrifugation, and then taking a supernatant, namely a biological sample extract; c) adding the same volume of a 3-nitrophenylhydrazine methanol solution and a 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide pyridine solution into the biological sample extract obtained in b), and performing uniform vortex mixing and heating for derivatization, wherein the concentration of used 3-nitrophenylhydrazine is 100-320 mmol/L, the concentration of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide is 50-200 mmol/L, the reaction temperature is 20-60 C., and the reaction time is 10-120 minutes; d) adding a carbon-13 labeled isotope internal standard solution obtained from the reaction of 3-nitrophenylhydrazine and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide into the derivatized biological sample extract obtained in c); and e) adding a methanol-water mixed solution into the sample in d) for dilution, and determining amino acid, phenol, phenyl or benzyl derivative, indole, organic acid, fatty acid, sugar, and bile acid by liquid chromatography-mass spectrometry.

11. The detection method according to claim 10, wherein in step c), the concentration of used 3-nitrophenylhydrazine is 150-220 mmol/L, the concentration of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide is 80-120 mmol/L, the reaction temperature is 20-40 C., and the reaction time is 30-60 minutes.

12. The detection method according to claim 10, wherein when the biological sample is urine, blood, saliva or cerebrospinal fluid, a treatment method of the biological sample in step b) comprises: taking an appropriate amount of the biological sample, extracting the biological sample with a mixed solvent of cold methanol, chloroform and water at a volume ratio of 3:1:1, shaking the mixture for a few seconds, performing centrifugation on the sample at a rotation speed of 10000-20000 rpm at a low temperature for 5-15 minutes, and transferring a supernatant into an autosampler glass vial for subsequent derivatization treatment.

13. The detection method according to claim 10, wherein when the biological sample is a fecal sample, a treatment method of the biological sample in step b) comprises: freeze-drying the fecal sample; homogenizing an appropriate amount of the freeze-dried fecal sample with an appropriate amount of a mixed solvent of cold methanol, chloroform and water at a volume ratio of 3:1:1, performing centrifugation on the sample at a rotation speed of 10000-20000 rpm at a low temperature for 15-30 minutes, and transferring a supernatant into an autosampler glass vial for subsequent derivatization treatment.

14. The detection method according to claim 10, wherein when the biological sample is a tissue or cell sample, a treatment method of the biological sample in step b) comprises: adding an appropriate amount of a mixed solvent of cold methanol, chloroform and water at a volume ratio of 3:1:1 into the sample for homogenization, performing centrifugation on the sample at a rotation speed of 10000-20000 rpm at 4 C. for 15-30 minutes, and transferring a supernatant into an autosampler glass vial for subsequent derivatization treatment.

15. The detection method according to claim 10, wherein the volume ratio of methanol to water in step e) is 1:1.

16. The detection method according to claim 1, wherein the multiple metabolic components of the biological sample detected by the quantitative detection method are selected from the following multiple metabolites: fructose 1,6-diphosphate, 10Z-nonadecenoic acid, trans-11-octadecenoic acid, cis-11-octadecenoic acid, 12-dehydrocholic acid, 12-hydroxystearic acid, 12-ketolithocholic acid, 1-methylhistidine, 2,2-dimethylsuccinic acid, 2,3-diaminopropionic acid, glucoside 24-chenodeoxycholic acid, 2-butenoic acid, 2-oxoadipic acid, 2-methyl-4-pentenoic acid, 2-methyl--alanine, 2-methylbutyric acid, 2-methylhexanoic acid, 2-methylglutaric acid, 2-methylglutamic acid, 2-furoic acid, 2-hydroxy-2-methylbutyric acid, 2-hydroxy-3-methylbutyric acid, 2-hydroxybutyric acid, 2-hydroxycaproic acid, 2-hydroxycinnamic acid, 2-hydroxyphenylacetic acid, 2-hydroxyhippuric acid, 2-phenylpropionic acid, 2-phenylglycine, 3-(3-hydroxyphenyl)-3-hydroxypropionic acid, 3-(4-hydroxyphenyl)lactic acid, 3,4-dihydroxymandelic acid, 3,4-dihydroxyphenylpropionic acid, 3,4-dehydro-DL-proline, 3,5-diiodo-L-thyroxine, 30-cholic acid, 3-pyridineacetic acid, 3-indoleacrylic acid, 3-indolepropionic acid, 3-indoleacetamide, 3-oxyalanine, 3-aminosalicylic acid, 3-chloro-L-tyrosine, 3-methyl-2-oxobutyric acid, 3-methyl-2-oxovaleric acid, 3-methylindole, 3-methyladipic acid, 3-methylglutamic acid, 3-nitrotyrosine, sulfate 3-taurolithocholic acid, sulfate 3-lithocholic acid, 3-hydroxy-2-aminobenzoic acid, 3-hydroxybutyric acid, 3-hydroxypropionic acid, 3-hydroxyisovaleric acid, 3-hydroxyphenylacetic acid, 3-hydroxyhippuric acid, 3-dehydrocholic acid, 3-phenylbutyric acid, 4-methyl-2-oxovaleric acid, 4-methylhexanoic acid, 4-methoxyphenylacetic acid, 4-hydroxy-3-methoxymandelic acid, 4-hydroxycinnamic acid, 4-hydroxybenzoic acid, 4-hydroxyhippuric acid, 4-hydroxyphenyllactic acid, 4-hydroxyphenylpyruvic acid, 4-phenylbutyric acid, 5-aminolevulinic acid, 5-hydroxytryptophan, 5-hydroxytryptamine, 5-hydroxylysine, 6,7-diketolithocholic acid, 6-phosphogluconic acid, 6-ketolithocholic acid, 7,12-diketolithocholic acid, 7-ketolithocholic acid, 7-ketodeoxycholic acid, 9,11-conjugated linoleic acid, D-2-hydroxyglutaric acid, D-galactose, D-xylose, D-xylulose, D-fructose, D-ribose, D-ribose-5-phosphate, D-ribulose, D-ribulose-5-phosphate, D-mannose, D-glucose, D-maltose, L-2-aminobutyric acid, L-3-phenyllactic acid, L-alanine, L-serine, L-acetylcarnitine, L-lactic acid, L-allothreonine, L-cysteine, L-homoserine, L-homocysteine, L-homocitrulline, L-pipecolic acid, L-aspartic acid, L-asparagine, L-sorbose, L-lignic acid, L-norleucine, L-kynurenine, L-thyronine, L-arginine, L-histidine, L-valine, L-cystine, L-cystathionine, L-proline, L-tryptophan, L-threonine, L-phenylalanine, L-malic acid, L-methionine, L-glutamine, L-glutamic acid, L-lysine, L-tyrosine, L-arabinose, N-(3-phenylpropionyl)glycine, N-acetyl-L-alanine, N-acetyl-L-aspartic acid, N-acetyl-L-phenylalanine, N-acetyl-L-methionine, N-acetyl-L-tyrosine, N-acetylserine, N-acetyl-D-glucosamine, N-acetyl-L-phenylalanine, N-acetylmannosamine, N-acetylneuraminic acid, N-acetylhistidine, N-acetylhydroxytryptamine, N-acetyltryptophan, N-acetylglutamine, N-acetyllysine, N-acetylornithine, N-methylnicotinamide, N-phenylacetyl-glutamine, N-phenylacetylphenylalanine, S-adenosine homocysteine, -D-glucose, -lactose, -linolenic acid, -hydroxyisobutyric acid, -ketoglutaric acid, -muricholic acid, -D-trehalose, -alanine, -ursocholic acid, -muricholic acid, -L-glutamyl-L-alanine, -linolenic acid, -aminobutyric acid, -muricholic acid, butyric acid, trimethylamine nitroxide, adenosine triphosphate, malonic acid, propionic acid, acetoacetic acid, acetylcysteine, acetylglycine, acetylornithine, acetic acid, guanidine acetate, glycolic acid, lactulose, lactoylglutathione, heneicosanoic acid, cis-12-heneicosenoic acid, heptacosanoic acid, tricosanoic acid, cis-14-tricosenoic acid, docosanoic acid, docosatrienoic acid, cis-13,16-docosadienoic acid, docosapentaenoic acid, docosahexaenoic acid, docosatetraenoic acid, trans-13-docosaenoic acid, cis-13-docosaenoic acid, pentacosanoic acid, octacosanoic acid, hexacosanoic acid, tetracosanoic acid, eicosanoic acid, eicosatrienoic acid, eicosadienoic acid, eicosapentaenoic acid, trans-11-eicosenoic acid, cis-11-eicosenoic acid, cis-5-eicosenoic acid, cis-8-eicosenoic acid, dimethylglycine, adenosine diphosphate, linoleic acid, leucine, alloisoleucine, allolithocholic acid, allocholic acid, undecenoic acid, heptadecanoic acid, tridecanoic acid, nonadecanoic acid, nonadecadienoic acid, pentadecanoic acid, galactonic acid, galactitol, mecysteine, protocatechuic acid, apocholic acid, dehydrolithocholic acid, nordeoxycholic acid, trans-9-tetradecenoic acid, trans-aconitic acid, trans-4-hydroxyproline, trans-9-heptadecenoic acid, trans-9-pentadecenoic acid, trans-9-hexadecenoic acid, trans-linolenic acid, trans-cinnamic acid, elaidic acid, homocysteine, pyrrole-2-carboxylic acid, picolinic acid, indole, indole-3-methyl acetate, indole-3-carboxylic acid, indoleacetic acid, purine, azelaic acid, nonanoic acid, dopa, dopamine, melibiose, fumaric acid, acetaminophen, p-aminohippuric acid, p-cresol sulfate, symmetrical dimethylarginine, p-hydroxymandelic acid, p-hydroxyphenylacetic acid, homogentisic acid, adipic acid, pimelic acid, isobutyric acid, isoleucine, isovaleric acid, citric acid, isoursodeoxycholic acid, isohyodeoxycholic acid, isolithocholic acid, isocholic acid, isodeoxycholic acid, glutaric acid, glutaconic acid, valeric acid, mandelic acid, lauric acid, fructose-6-phosphate, citraconic acid, citric acid, citramalic acid, ribonolactone, ribonic acid, raffinose, palmitoleic acid, palmitic acid, norvaline, n-hydroxyphenylacetic acid, oxidized glutathione, aminoadipic acid, aminocaproic acid, salicyluric acid, oleic acid, trehalose, nicotinic acid, pyroglutamic acid, ursocholic acid, ursodeoxycholic acid, tauro-a-muricholic acid, tauro-b-muricholic acid, tauro-w-muricholic acid, tauroursodeoxycholic acid, taurohyocholic acid, taurohyodeoxycholic acid, taurolithocholic acid, taurocholic acid, taurodehydrocholic acid, taurochenodeoxycholic acid, hyocholic acid, hyodeoxycholic acid, succinylacetone, succinic acid, citrulline, glycodehydrocholic acid, glycolithocholic acid, glycodeoxycholic acid, glycine, glycochenodeoxycholic acid, glyceraldehyde, glyceraldehyde-3-phosphate, choline glycerophosphate, glycoursodeoxycholic acid, glycohyocholic acid, glycohyodeoxycholic acid, glycocholic acid, glyproline, glycyl-L-leucine, mannose-6-phosphoric acid, mannitol, methylmalonic acid, methylsuccinic acid, formic acid, capric acid, lithocholic acid, selenomethionine, thiamine, glycolithocholic sulfate, stearic acid, liothyronine, dihydroxyacetone phosphate, phosphoribosyl pyrophosphate, creatine phosphate, hydroxypyruvic acid, glycine hydroxyphenylacetate, cinnamic acid, sarcosine, carnosine, creatine, inositol, epinephrine, choline, cholic acid, dehydrocholic acid, demethylcholic acid, adenosine monophosphate, arachidonic acid, phenylpyruvic acid, phenethylamine, phenylpropionic acid, phenyllactic acid, benzamide, benzoic acid, oxalic acid, shikimic acid, glucaric acid, glucose-6-phosphate, glucose lactone, sebacic acid, ricinoleic acid, sucrose, methionine sulfoxide, itaconic acid, melatonin, glutathione, myristic acid, erythronic acid, erythrose, suberic acid, caprylic acid, phthalic acid, tartaric acid, tyramine, quinic acid, m-aminobenzoic acid, asymmetric dimethylarginine, tannic acid, cis-10,12-octadecadienoic acid, cis-12,15-heneicosadienoic acid, cis-12-tridecenoic acid, cis-15-tetracosenic acid, cis-2-hydroxycinnamic acid, cis-9-tetradecenoic acid, cis-10-heptadecenoic acid, cis-11-dodecenoic acid, cis-4-hydroxyproline, cis-5-dodecenoic acid, cis-7-hexadecenoic acid, cis-9-heptadecenoic acid, cis-aconitic acid, vanillic acid, hippuric acid, maleic acid, homovanillic acid, ornithine, guanosine monophosphate, guanosine triphosphate, anserine, chenodeoxycholic acid, maltotriose, maltitol, rhamnose and murideoxycholic acid.

17. A metabolic chip used in the detection method according to claim 1, comprising a chip carrier, a filter device and dry solid powder of a standard product and a quality control product, wherein the chip carrier is a microtiter plate, each well of the microtiter plate is provided with an independent filter device, and the powder obtained by dehydrating or freeze-drying solutions of the standard product and the quality control product is placed on the filter device in each well of the microtiter plate; the multiple metabolic components in the standard product are selected from amino acid, phenol, phenyl or benzyl derivative, indole, organic acid, fatty acid, sugar, and bile acid standard products; the multiple metabolic components in the quality control product are selected from amino acid, phenol, phenyl or benzyl derivative, indole, organic acid, fatty acid, sugar, and bile acid standard products corresponding to the standard products above; the number of the multiple metabolic components in the standard product and the quality control product is 50 or more.

18. The metabolic chip according to claim 17, wherein the microtiter plate is selected from a 48-well plate, a 96-well plate and a 384-well plate, and the filter device is a filter membrane made of polyvinylidene fluoride, cellulose acetate, or nylon with a pore size of 0.20-0.45 micron.

19. The metabolic chip according to claim 17, wherein the multiple metabolic components in the standard product and the quality control product are selected from: fructose 1,6-diphosphate, 10Z-nonadecenoic acid, trans-11-octadecenoic acid, cis-11-octadecenoic acid, 12-dehydrocholic acid, 12-hydroxystearic acid, 12-ketolithocholic acid, 1-methylhistidine, 2,2-dimethylsuccinic acid, 2,3-diaminopropionic acid, glucoside 24-chenodeoxycholic acid, 2-butenoic acid, 2-oxoadipic acid, 2-methyl-4-pentenoic acid, 2-methyl--alanine, 2-methylbutyric acid, 2-methylhexanoic acid, 2-methylglutaric acid, 2-methylglutamic acid, 2-furoic acid, 2-hydroxy-2-methylbutyric acid, 2-hydroxy-3-methylbutyric acid, 2-hydroxybutyric acid, 2-hydroxycaproic acid, 2-hydroxycinnamic acid, 2-hydroxyphenylacetic acid, 2-hydroxyhippuric acid, 2-phenylpropionic acid, 2-phenylglycine, 3-(3-hydroxyphenyl)-3-hydroxypropionic acid, 3-(4-hydroxyphenyl)lactic acid, 3,4-dihydroxymandelic acid, 3,4-dihydroxyphenylpropionic acid, 3,4-dehydro-DL-proline, 3,5-diiodo-L-thyroxine, 30-cholic acid, 3-pyridineacetic acid, 3-indoleacrylic acid, 3-indolepropionic acid, 3-indoleacetamide, 3-oxyalanine, 3-aminosalicylic acid, 3-chloro-L-tyrosine, 3-methyl-2-oxobutyric acid, 3-methyl-2-oxovaleric acid, 3-methylindole, 3-methyladipic acid, 3-methylglutamic acid, 3-nitrotyrosine, sulfate 3-taurolithocholic acid, sulfate 3-lithocholic acid, 3-hydroxy-2-aminobenzoic acid, 3-hydroxybutyric acid, 3-hydroxypropionic acid, 3-hydroxyisovaleric acid, 3-hydroxyphenylacetic acid, 3-hydroxyhippuric acid, 3-dehydrocholic acid, 3-phenylbutyric acid, 4-methyl-2-oxyvaleric acid, 4-methylhexanoic acid, 4-methoxyphenylacetic acid, 4-hydroxy-3-methoxymandelic acid, 4-hydroxycinnamic acid, 4-hydroxybenzoic acid, 4-hydroxyhippuric acid, 4-hydroxyphenyllactic acid, 4-hydroxyphenylpyruvic acid, 4-phenylbutyric acid, 5-aminolevulinic acid, 5-hydroxytryptophan, 5-hydroxytryptamine, 5-hydroxylysine, 6,7-diketolithocholic acid, 6-phosphogluconic acid, 6-ketolithocholic acid, 7,12-diketolithocholic acid, 7-ketolithocholic acid, 7-ketodeoxycholic acid, 9,11-conjugated linoleic acid, D-2-hydroxyglutaric acid, D-galactose, D-xylose, D-xylulose, D-fructose, D-ribose, D-ribose-5-phosphate, D-ribulose, D-ribulose-5-phosphate, D-mannose, D-glucose, D-maltose, L-2-aminobutyric acid, L-3-phenyllactic acid, L-alanine, L-serine, L-acetylcarnitine, L-lactic acid, L-allothreonine, L-cysteine, L-homoserine, L-homocysteine, L-homocitrulline, L-pipecolic acid, L-aspartic acid, L-asparagine, L-sorbose, L-lignic acid, L-norleucine, L-kynurenine, L-thyronine, L-arginine, L-histidine, L-valine, L-cystine, L-cystathionine, L-proline, L-tryptophan, L-threonine, L-phenylalanine, L-malic acid, L-methionine, L-glutamine, L-glutamic acid, L-lysine, L-tyrosine, L-arabinose, N-(3-phenylpropionyl)glycine, N-acetyl-L-alanine, N-acetyl-L-aspartic acid, N-acetyl-L-phenylalanine, N-acetyl-L-methionine, N-acetyl-L-tyrosine, N-acetylserine, N-acetyl-D-glucosamine, N-acetyl-L-phenylalanine, N-acetylmannosamine, N-acetylneuraminic acid, N-acetylhistidine, N-acetylhydroxytryptamine, N-acetyltryptophan, N-acetylglutamine, N-acetyllysine, N-acetylornithine, N-methylnicotinamide, N-phenylacetyl-glutamine, N-phenylacetylphenylalanine, S-adenosine homocysteine, -D-glucose, -lactose, -linolenic acid, -hydroxyisobutyric acid, -ketoglutaric acid, -muricholic acid, -D-trehalose, -alanine, -ursocholic acid, -muricholic acid, -L-glutamyl-L-alanine, -linolenic acid, -aminobutyric acid, -muricholic acid, butyric acid, trimethylamine nitroxides, adenosine triphosphate, malonic acid, propionic acid, acetoacetic acid, acetylcysteine, acetylglycine, acetylornithine, acetic acid, guanidine acetate, glycolic acid, lactulose, lactoylglutathione, heneicosanoic acid, cis-12-heneicosenoic acid, heptacosanoic acid, tricosanoic acid, cis-14-tricosenoic acid, docosanoic acid, docosatrienoic acid, cis-13,16-docosadienoic acid, docosapentaenoic acid, docosahexaenoic acid, docosatetraenoic acid, trans-13-docosaenoic acid, cis-13-docosaenoic acid, pentacosanoic acid, octacosanoic acid, hexacosanoic acid, tetracosanoic acid, eicosanoic acid, eicosatrienoic acid, eicosadienoic acid, eicosapentaenoic acid, trans-11-eicosenoic acid, cis-11-eicosenoic acid, cis-5-eicosenoic acid, cis-8-eicosenoic acid, dimethylglycine, adenosine diphosphate, linoleic acid, leucine, alloisoleucine, allolithocholic acid, allocholic acid, undecenoic acid, heptadecanoic acid, tridecanoic acid, nonadecanoic acid, nonadecadienoic acid, pentadecanoic acid, galactonic acid, galactitol, mecysteine, protocatechuic acid, apocholic acid, dehydrolithocholic acid, nordeoxycholic acid, trans-9-tetradecenoic acid, trans-aconitic acid, trans-4-hydroxyproline, trans-9-heptadecenoic acid, trans-9-pentadecenoic acid, trans-9-hexadecenoic acid, trans-linolenic acid, trans-cinnamic acid, elaidic acid, homocysteine, pyrrole-2-carboxylic acid, picolinic acid, indole, indole-3-methyl acetate, indole-3-carboxylic acid, indoleacetic acid, purine, azelaic acid, nonanoic acid, dopa, dopamine, melibiose, fumaric acid, acetaminophen, p-aminohippuric acid, p-cresol sulfate, symmetrical dimethylarginine, p-hydroxymandelic acid, p-hydroxyphenylacetic acid, homogentisic acid, adipic acid, pimelic acid, isobutyric acid, isoleucine, isovaleric acid, citric acid, isoursodeoxycholic acid, isohyodeoxycholic acid, isolithocholic acid, isocholic acid, isodeoxycholic acid, glutaric acid, glutaconic acid, valeric acid, mandelic acid, lauric acid, fructose-6-phosphate, citraconic acid, citric acid, citramalic acid, ribonolactone, ribonic acid, raffinose, palmitoleic acid, palmitic acid, norvaline, n-hydroxyphenylacetic acid, oxidized glutathione, aminoadipic acid, aminocaproic acid, salicyluric acid, oleic acid, trehalose, nicotinic acid, pyroglutamic acid, ursocholic acid, ursodeoxycholic acid, tauro-a-muricholic acid, tauro-b-muricholic acid, tauro-w-muricholic acid, tauroursodeoxycholic acid, taurohyocholic acid, taurohyodeoxycholic acid, taurolithocholic acid, taurocholic acid, taurodehydrocholic acid, taurochenodeoxycholic acid, hyocholic acid, hyodeoxycholic acid, succinylacetone, succinic acid, citrulline, glycodehydrocholic acid, glycolithocholic acid, glycodeoxycholic acid, glycine, glycochenodeoxycholic acid, glyceraldehyde, glyceraldehyde-3-phosphate, choline glycerophosphate, glycoursodeoxycholic acid, glycohyocholic acid, glycohyodeoxycholic acid, glycocholic acid, glyproline, glycyl-L-leucine, mannose-6-phosphoric acid, mannitol, methylmalonic acid, methylsuccinic acid, formic acid, capric acid, lithocholic acid, selenomethionine, thiamine, glycolithocholic sulfate, stearic acid, liothyronine, dihydroxyacetone phosphate, phosphoribosyl pyrophosphate, creatine phosphate, hydroxypyruvic acid, glycine hydroxyphenylacetate, cinnamic acid, sarcosine, carnosine, creatine, inositol, epinephrine, choline, cholic acid, dehydrocholic acid, demethylcholic acid, adenosine monophosphate, arachidonic acid, phenylpyruvic acid, phenethylamine, phenylpropionic acid, phenyllactic acid, benzamide, benzoic acid, oxalic acid, shikimic acid, glucaric acid, glucose-6-phosphate, glucose lactone, sebacic acid, ricinoleic acid, sucrose, methionine sulfoxide, itaconic acid, melatonin, glutathione, myristic acid, erythronic acid, erythrose, suberic acid, caprylic acid, phthalic acid, tartaric acid, tyramine, quinic acid, m-aminobenzoic acid, asymmetric dimethylarginine, tannic acid, cis-10,12-octadecadienoic acid, cis-12,15-heneicosadienoic acid, cis-12-tridecenoic acid, cis-15-tetracosenic acid, cis-2-hydroxycinnamic acid, cis-9-tetradecenoic acid, cis-10-heptadecenoic acid, cis-11-dodecenoic acid, cis-4-hydroxyproline, cis-5-dodecenoic acid, cis-7-hexadecenoic acid, cis-9-heptadecenoic acid, cis-aconitic acid, vanillic acid, hippuric acid, maleic acid, homovanillic acid, ornithine, guanosine monophosphate, guanosine triphosphate, anserine, chenodeoxycholic acid, maltotriose, maltitol, rhamnose and murideoxycholic acid.

20. A use method of the metabolic chip according to claim 17, comprising the following steps: a) collecting a biological sample; b) according to the sample type, preparing a corresponding biological sample extract, when the biological sample is urine, blood, saliva or cerebrospinal fluid, a treatment method of the biological sample comprises: taking an appropriate amount of the biological sample, extracting the biological sample with a mixed solvent of cold methanol, chloroform and water at a volume ratio of 3:1:1, shaking the mixture for a few seconds, performing centrifugation on the sample at a rotation speed of 10000-20000 rpm at a low temperature for 5-15 minutes, and transferring a supernatant into an autosampler glass vial for subsequent derivatization treatment; c) adding the prepared biological sample extract into each well of the metabolic chip in an equal amount, adding the same volume of a 3-nitrophenylhydrazine methanol solution and a 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide pyridine solution into each well, and performing uniform vortex mixing and heating for derivatization, wherein the concentration of used 3-nitrophenylhydrazine is 100-320 mmol/L, the concentration of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide is 50-200 mmol/L, the reaction temperature is 20-60 C., and the reaction time is 10-120 minutes; d) adding a carbon-13 labeled isotope internal standard solution obtained from the reaction of 3-nitrophenylhydrazine and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide into the derivatized biological sample extract obtained in c); and e) adding a methanol-water mixed solution into each well in the metabolic chip in d) for dilution, placing the metabolic chip in a tandem mass spectrometer for determination of amino acid, phenol, phenyl or benzyl derivative, indole, organic acid, fatty acid, sugar, and bile acid by liquid chromatography-mass spectrometry, and calculating concentrations of target metabolites in the sample based on results.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0059] The accompanying drawings are chromatograms of typical amino acid, organic acid, fatty acid, sugar, and bile acid detected in blood samples in embodiments. It can be seen that multiple target detection components can be effectively separated and detected, and thus high-throughput quantitative detection is achieved.

[0060] FIG. 1 is a chromatogram of typical short-chain fatty acid;

[0061] FIG. 2 is a chromatogram of typical amino acid;

[0062] FIG. 3 is a chromatogram of typical organic acid;

[0063] FIG. 4 is a chromatogram of typical sugar;

[0064] FIG. 5 is a chromatogram of typical bile acid.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

[0065] A metabolic chip is used to detect multiple index components in 10 human blood and fecal samples. FIGS. 1-5 are chromatograms of typical amino acid, organic acid, fatty acid, sugar, and bile acid detected in blood samples in this Example.

1. Instrument

[0066] Liquid chromatography-tandem mass spectrometer (LC-MS/MS) equipped with an electrospray ionization source (ESI).

2. Sample Preparation

[0067] Serum sample: Avenous whole blood sample is collected, placed in an anticoagulation tube, then immediately shaken up and down for uniform mixing 5-6 times, and centrifugated within 30 minutes to separate plasma. The sample is placed in a centrifuge tube, extracted with a mixed solvent of cold methanol, chloroform and water at a volume ratio of 3:1:1, shaken for a few seconds and then centrifugated at a rotation speed of 10000-20000 rpm at 4 C. for 5-15 minutes to obtain a supernatant. The supernatant is transferred into an autosampler glass vial. All water-containing serum or urine sample extracts are used for subsequent derivatization treatment.

[0068] Fecal sample: A fecal sample is freeze-dried. An appropriate amount of the freeze-dried fecal sample and an appropriate amount of a mixed solvent of cold methanol, chloroform, and water at a volume ratio of 3:1:1 are homogenized. The sample is centrifugated at a rotation speed of 10000-20000 rpm at 4 C. for 15-30 minutes to obtain a supernatant. The supernatant is transferred into an autosampler glass vial for subsequent derivatization treatment.

3. Reagent Preparation

[0069] Preparation of standard product solution: Standard products, including amino acid, phenol, phenyl or benzyl derivative, indole, organic acid, fatty acid, sugar, and bile acid, of metabolites in the description are taken, fully dissolved in methanol and uniformly mixed to prepare a 1 mg/ml solution, namely a concentrated stock solution which is prepared into a series of concentrations of solutions to draw a standard curve.

[0070] Preparation of quality control product solution: A corresponding quality control product is taken, fully dissolved in methanol and uniformly mixed to prepare a 1 mg/ml solution, namely a concentrated stock solution, and then the solution is diluted to a certain concentration and reacts with carbon-13 labeled 3-nitrophenylhydrazine and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide to obtain the quality control product solution.

[0071] Preparation of derivatization reagent (3-nitrophenylhydrazine): A derivatization reagent is uniformly mixed with a 75% methanol aqueous solution to prepare a 200 mM solution, and the solution is sealed and stored at 4 C. for later use.

[0072] Preparation of derivatization reaction catalyst (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide): A reaction catalyst is prepared into a 120 millimolar solution with pyridine, and the solution is sealed and stored at 4 C. for later use.

4. Inspection Method

[0073] 5 L of a treated biological sample is taken, 20 l of a derivatization reagent and 20 l of a derivatization catalyst are added for reaction at 30 C. for 60 minutes, a methanol-water mixed solution is added into a reaction solution for dilution, centrifugation is performed at 13200 rpm for 15 minutes, and 5 L of a supernatant is taken and introduced for LC-MS/MS analysis.

Mass Spectrometry Conditions:

[0074] Ion source: Multi-reaction detection conditions for detection of multiple substances by tandem mass spectrometry are shown in Table 1. A negative ion scanning mode (ESI) is adopted for an electrospray ion source, and specific conditions are as follows: The capillary voltage is 1.2 kV, the cone voltage is 55 V, the extraction cone voltage is 4 V, the ion source temperature is 150 C., the desolvent gas temperature is 550 C., the reverse cone gas flow is 50 L/h, the desolvent gas is 650 L/h, the resolution of a low mass zone is 4.7, the resolution of a high mass zone is 15, and a multi-reaction detection mode is used to collect data.

[0075] Gradient elution conditions: A UPLC BEH C18 chromatographic column (100 mm*2.1 mm, 1.7 m) is used; the column temperature is 40 C.; a mobile phase A includes water (0.1% formic acid), and a mobile phase B includes acetonitrile (0.1% formic acid) and isopropanol at a ratio of (1-2):1; the flow rate is 0.4 mL/min; the injection volume is 5 microliters; and gradient elution conditions: 0-1 min (5% B), 1-5 min (5-30% B), 5-9 min (30-50% B), 9-12 min (50-75% B), 12-15 min (75-95% B), 15-16 min (95-100% B), 16-18 min (100% B), 18-20 min (5% B).

5. Determination Results

[0076] Concentrations of substances in 10 human blood and fecal samples are detected by using the detection method of the present invention (see Table 1 and Table 2 respectively). It can be seen that by using the method of the present invention to detect a single sample, multiple substances of different magnitudes and different properties can be quantitatively detected at one time.

TABLE-US-00001 TABLE 1 Determination results of concentrations of substances in blood of normal people Concentration Concentration Determined target metabolites value unit 2-methylvaleric acid 120.32 5.18 g/mL 2-hydroxybutyric acid 2.32 0.44 g/mL 3-(3-hydroxyphenyl)-3- 10.31 0.19 g/mL hydroxypropionic acid 3-hydroxyphenylacetic acid 6.07 6.12 g/mL 3-indoleacetonitrile 2.51 0.15 g/mL 3-methyl-2-oxovaleric acid 4.37 5.08 g/mL 4-methylhexanoic acid 2.96 0.07 g/mL Linolenic acid 97.19 33.87 g/mL Arachidonic acid 5.54 0.45 g/mL Arachidonic acid 5.62 2.4 g/mL Docosanoic acid 8.96 0.27 g/mL -alanine 10.37 0.44 g/mL Capric acid 1.22 0.19 g/mL Caprylic acid 1.39 0.5 g/mL Citric acid 7.03 1.56 g/mL Docosahexaenoic acid 15.1 4.36 g/mL Docosapentaenoic acid n6 15.31 4.16 g/mL Docosatrienoic acid 8.76 0.95 g/mL Dodecanoic acid 0.91 0.16 g/mL Dopamine 23.53 3.93 g/mL Eicosenoic acid 11.18 3.38 g/mL Erucic acid 15.25 11.34 g/mL -aminobutyric acid 14.68 2.7 g/mL Glutathione 6.7 1.62 g/mL Glycolic acid 159.46 56.76 g/mL Heptadecanoic acid 5.78 6.5 g/mL L--aminobutyric acid 2.41 0.48 g/mL L-asparagine 12.32 0.57 g/mL L-aspartic acid 3.8 0.64 g/mL L-glutamic acid 8.93 0.53 g/mL L-histidine 16.76 1.54 g/mL L-homoserine 12.53 7.81 g/mL L-isoleucine 5.9 2.18 g/mL L-leucine 5.88 3.97 g/mL L-lysine 33.73 4.42 g/mL L-methionine 6.78 0.73 g/mL L-norleucine 3.33 1.25 g/mL L-phenylalanine 8.57 0.84 g/mL L-proline 11.32 5.94 g/mL L-serine 9.78 0.68 g/mL L-tryptophan 30.49 2.93 g/mL L-tyrosine 20.02 4.37 g/mL L-valine 28.87 7.87 g/mL Linoleic acid 162.6 57.21 g/mL Methylsuccinic acid 34.76 29.31 g/mL Myristic acid 2.91 0.52 g/mL Myristic acid 3.39 0.51 g/mL N-acetyltryptophan 29.5 1.24 g/mL Nervonic acid 82.71 4.32 g/mL Dodecanoic acid 5.9 2.13 g/mL Norvaline 0.86 0.01 g/mL Ornithine 22.56 5.54 g/mL Carbonyladipic acid 11.55 2.52 g/mL Oxoglutaric acid 21.89 5.22 g/mL Palmitic acid 87.29 30.74 g/mL Palmitoleic acid 86.36 30.14 g/mL Pentadecanoic acid 1.81 0.12 g/mL Pimelic acid 4.47 1.23 g/mL Propionic acid 0.16 0.03 g/mL Putrescine 19.96 1.32 g/mL Pyroglutamic acid 5.59 2 g/mL Stearic acid 38.06 14.04 g/mL Succinic acid 36.45 12.11 g/mL Cis-aconitic acid 2.66 0.11 g/mL P-hydroxyphenylacetic acid 2.51 0.15 g/mL Palmitoleic acid 19.32 11.42 uM Nervonic acid 0.25 0.06 uM Cholic acid 67.57 38.23 nM Chenodeoxycholic acid 383 559.28 nM Deoxycholic acid 241.97 197.98 nM Fructose 5.23 1.18 uM Glucose 3.29 0.66 mM Dohomo-g-linoleic acid 2.51 1.01 uM Mannose 34.87 9.22 uM Glycocholic acid 314.89 345.38 nM Glycochenodeoxycholic acid 750.99 574.2 nM Glycohyocholic acid 16.31 10.67 nM Glycolithocholic acid 12.41 10.41 nM Glycoursocholic acid 123.97 124.56 nM Hyocholic acid 26.09 11.26 nM Lithocholic acid 12.61 4.68 nM Oleic acid 239.05 84.26 uM Palmitoleic acid 19.32 11.42 uM Taurocholic acid 88.59 66.4 nM Taurodeoxycholic acid 68.66 47.85 nM Teracosanoic acid 2.28 0.89 uM Taurohyocholic acid 4.06 4.7 nM Tauroursocholic acid 26.16 0.81 nM Ursocholic acid 69.29 44.3 nM g-linoleic acid 20.47 8.36 uM

TABLE-US-00002 TABLE 2 Determination results of concentrations of substances in feces of normal people Concentration Concentration Detected target metabolites value unit (1)-2-methylvaleric acid 0.1 0.19 g/mL 1H-indole-3-acetamide 0.54 0.07 g/mL 2-hydroxybutyric acid 29.5 7.89 ng/mL 3-(3-hydroxyphenyl)-3- 0.19 0.11 g/mL hydroxypropionic acid 3-hydroxybutyric acid 0.3 0.29 g/mL 3-hydroxyphenylacetic acid 0.39 0.36 g/mL 3-indoleacetonitrile 0.17 0.11 g/mL 3-isopropionic acid 0.32 0.09 g/mL 3-methyl-2-oxovaleric acid 0.5 0.2 ng/mL 3-methylvaleric acid 0.2 0.3 ng/mL 4-hydroxybenzoic acid 0.42 0.55 g/mL 4-hydroxycinnamic acid 0.25 0.07 g/mL 4-methylhexanoic acid 0.7 0.6 ng/mL 5-dodecenoic acid 0.84 2.1 g/mL Adipic acid 88.6 10.6 ng/mL -linolenic acid 8.07 9.95 g/mL Aminoadipic acid 0.14 0.04 g/mL Arachidonic acid 3.83 2.82 g/mL Arachidonic acid 0.97 0.88 g/mL Docosanoic acid 0.33 0.32 g/mL -alanine 0.41 0.12 g/mL Butyric acid 0.88 0.69 g/mL Capric acid 0.13 0.22 g/mL Caproic acid 0.53 0.53 g/mL Caprylic acid 0.37 0.55 g/mL Citraconic acid 45.8 4.97 ng/mL 2-methylmalic acid 0.18 0.07 g/mL Citric acid 0.34 0.32 g/mL D-2-hydroxyglutaric acid 0.14 0.06 g/mL Docosahexaenoic acid 0.43 0.3 g/mL Docosapentaenoic acid n6 0.45 0.41 g/mL Docosatrienoic acid 0.16 0.02 g/mL Dodecanoic acid 1.07 2.33 g/mL Eicosenoic acid 8.91 6.15 g/mL Erucic acid 0.41 0.36 g/mL Ethylmethylacetic acid 0.73 0.38 g/mL Fumaric acid 0.12 0.04 g/mL -aminobutyric acid 0.81 0.37 g/mL Glutaric acid 0.32 0.27 g/mL Glutathione 3.33 5.08 g/mL Glyceric acid 4.11 2.07 g/mL Glycolic acid 0.27 0.21 g/mL Heptadecanoic acid 1.06 1.15 g/mL Heptanoic acid 0.12 0.15 g/mL Homocysteine l.l 1.01 g/mL Hydrocinnamic acid 0.36 0.21 g/mL Hydroxyphenyllactic acid 0.48 0.27 g/mL Hydroxypropionic acid 0.17 0.2 g/mL Indole 0.4 0.34 g/mL Indoleacetic acid 0.29 0.02 g/mL Isocitric acid 89.5 5.67 ng/mL Itaconic acid 92.4 15.4 ng/mL L--aminobutyric acid 74.7 16.7 ng/mL L-asparagine 0.88 0.59 g/mL L-aspartic acid 2.44 1.03 g/mL L-glutamic acid 10.35 8.3 g/mL L-histidine 0.39 0.07 g/mL L-homoserine 0.45 0.42 g/mL L-isoleucine 0.23 0.11 g/mL L-leucine 0.25 0.42 g/mL L-lysine 2.29 0.65 g/mL L-methionine 1.54 0.91 g/mL L-norleucine 0.13 0.25 g/mL L-phenylalanine 0.85 0.91 g/mL L-proline 0.4 0.21 g/mL L-serine 0.42 0.31 g/mL L-tryptophan 0.62 0.16 g/mL L-tyrosine 2.19 1.06 g/mL L-valine 1.05 0.6 g/mL Linoleic acid 0.012 0.05 mg/mL Malic acid 0.38 0.21 g/mL Malonic acid 0.12 0.06 g/mL Methylsuccinic acid 0.13 0.02 g/mL Myristic acid 0.54 0.54 g/mL Myristic acid 0.53 0.53 g/mL N-acetyl 0.61 0.07 g/mL Nervonic acid 1.33 0.9 g/mL Nicotinic acid 0.31 0.09 g/mL Dodecanoic acid 0.21 0.11 g/mL Norvaline 29.5 4.56 ng/mL Ornithine 1.29 1.12 g/mL Oxalic acid 3.49 5.07 g/mL Carbonyladipic acid 0.72 0.41 g/mL Oxoglutaric acid 0.17 0.17 g/mL Palmitic acid 3.79 2.29 g/mL Palmitoleic acid 3.69 2.19 g/mL Nonanoic acid 87.4 12.5 ng/mL Pentadecanoic acid 0.31 0.24 g/mL Phenol 58.7 6.78 ng/mL Phenylacetic acid 1.49 1.14 g/mL Phenyllactic acid 71.5 8.79 ng/mL Pimelic acid 0.31 0.22 g/mL Propionic acid 10.09 6.03 g/mL Pyroglutamic acid 2.6 3.95 g/mL Stearic acid 1.22 1.63 g/mL Suberic acid 0.11 0.03 g/mL Succinic acid 0.42 0.32 g/mL Tartaric acid 80.3 9.97 ng/mL Thiamine 2.23 4.06 g/mL Valeric acid 1.39 1.36 g/mL Vanillic acid 0.17 0.04 g/mL Cis-aconitic acid 70.05 10.08 ng/mL P-cresol 0.24 0.19 g/mL P-hydroxyphenylacetic acid 0.3 0.22 g/mL

Example 2

[0077] A metabolic chip is used to detect multiple index components in 6 human blood and fecal samples.

1. Reagent Preparation

[0078] Preparation of standard product solution: Standard products, including amino acid, phenol, phenyl or benzyl derivative, indole, organic acid, fatty acid, sugar, and bile acid, of metabolites in the description are taken, fully dissolved in methanol and uniformly mixed to prepare a 1 mg/ml solution, namely a concentrated stock solution which is prepared into a series of concentrations of solutions to draw a standard curve.

[0079] Preparation of derivatization reagent (3-nitrophenylhydrazine): A derivatization reagent is uniformly mixed with a 75% methanol aqueous solution to prepare a 200 mM solution, and the solution is sealed and stored at 4 C. for later use.

[0080] Preparation of derivatization reaction catalyst (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide): A reaction catalyst is prepared into a 120 millimolar solution with pyridine, and the solution is sealed and stored at 4 C. for later use.

2. Sample Preparation

[0081] Preparation of serum extraction: An aliquot of standard solution or 20 L of serum was mixed with 120 L of methanol. Centrifugation at 13,500 g and 4 C. for 10 min.

[0082] Preparation of feces extraction: Feces samples were freeze-dried, and approximately 5 mg of each sample was weighed. The samples were homogenized with 20 L of ultrapure water and extracted with 120 L of methanol. Centrifugation at 13,500 g and 4 C. for 10 min. L of a treated biological supernatant is taken, 20 l of a derivatization reagent and 20 l of a derivatization catalyst are added for reaction at 30 C. for 60 minutes, a methanol-water mixed solution is added into a reaction solution for dilution, centrifugation is performed at 13200 rpm for 15 minutes, and 5 L of a supernatant is taken and introduced for LC-MS/MS analysis.

3. Inspection Method

UHPLC/Q-Orbitrap-MS Analysis

[0083] High-resolution mass spectrometry (HRMS) data for multicomponent characterization and untargeted metabolomics were acquired using a Vanquish Flex UHPLC system coupled to a Q Extractive Focus Orbitrap mass spectrometer equipped with a heated electrospray ionization (ESI) source (Thermo Fisher Scientific, San Jose, CA, USA). Chromatographic separation was performed on a reversed-phase ACQUITY UPLC BEH C18 column (2.1 mm100 mm, 1.7 m particle size) using a binary gradient elution with mobile phases: 0.1% formic acid in water (solvent A) and acetonitrile containing 0.1% formic acid/isopropanol (70:30, v/v, solvent B). The column temperature is 40 C. The flow rate is 0.4 mL/min; the injection volume is 5 microliters. The gradient program was as follows: 0-1 min (5% B), 1-5 min (5-30% B), 5-9 min (30-50% B), 9-12 min (50-75% B), 12-15 min (75-95% B), 15-16 min (95-100% B), 16-18 min (100% B), 18-20 min (5% B).

[0084] Mass Spectrometry Parameters: The mass spectrometry scan parameters were set as follows: S-lens RF level, 50; mass range, 100 to 1200 m/z; full MS resolution, 70,000; MS/MS resolution, 17,500; NCE/stepped NCE was set at 10, 20, and 40 eV. The ESI ion source parameters were set as follows: spray voltage, 3800 V; sheath gas flow rate, 40 for ESI+ and 5 for ESI; capillary temperature, 320 C.; probe heater temperature, 350 C.; aux gas flow rate, 10.

4. Determination Results

[0085] Concentrations of substances in 6 human serum and fecal samples are detected by using the detection method of the present invention (Table 3). It can be seen that by using the method of the present invention to detect a single sample, multiple substances of different magnitudes and different properties can be quantitatively detected at one time.

TABLE-US-00003 TABLE 3 Determination results of concentrations of substances in blood and feces of normal people RT Reference Fecal Serum NO. Name m/z (min) Ion (Intensity) (Intensity) 1 Propynoic acid 264.06 6.246 [M H + HAc] 1 2510269 841952 92554 28455 2 Acrylic acid 208.07 0.883 [M + H] + 1 13865154 14228426 12993928 23777442 3 Glyoxylic acid 208.04 5.832 [M H] 1 12000471 2144384 2000381 228813 4 Isocrotonic acid 239.11 0.434 [M + NH4] + 1 266059225 218886 460019188 43040 5 2-Aminoacrylic acid 221.07 6.247 [M H] 1 3464018 6160226 800230 1903288 6 Malonic semialdehyde 222.05 9.361 [M H] 1 8953281 1962295 3833020 1202549 7 2-hydroxyacrylic Acid 222.05 7.364 [M H] 1 40949997 9472056 4052812 1132094 8 (alpha-D-mannosyl)7-beta-D-mannosyl- 223.08 1.516 [M H] 1 458487665 148636884 diacetylchitobiosyl-L-asparagine, 63646212 13164304 isoform A (protein) 9 Monoethyl carbonate 224.07 4.177 [M H] 1 246941092 71459981 7837139 3414768 10 Methoxyacetic acid 224.07 3.726 [M H] 1 5184592776 2172177007 72558533 122437118 11 2-hydroxypropanoic acid 224.07 5.824 [M H] 1 157271490 8346423 13475327 299209 12 Senecioic acid 236.10 2.004 [M + H] + 1 76683919 10848175 56264980 10483810 13 Tiglic acid 253.13 0.6 [M + NH4] + 1 1509117642 58052130 2611743446 35789141 14 2-Ethylacrylic acid 253.13 1.254 [M + NH4] + 1 1862909805 2059982 1650227504 695343 15 Angelic acid 253.13 1.784 [M + NH4] + 1 50362708 9238657 86502238 4771737 16 2-Ketobutyric acid 236.07 7.196 [M H] 1 1787233 3482001 91095 1733281 17 Succinic acid semialdehyde 236.07 4.368 [M H] 1 16202483 4515125 4170048 168205 18 4-Hydroxycrotonic acid 236.07 8.057 [M H] 1 142453769 31505827 11877298 3548011 19 (S)-2-Methylbutanoic acid 236.10 6.982 [M H] 1 3463810327 7900237 108796706 1585336 20 Pivalic acid 236.10 7.443 [M H] 1 6972066992 8738812 105310392 1653469 21 Pyruvatoxime 237.06 2.504 [M H] 1 5796134 248976 360009 125681 22 (R)-beta-Aminoisobutyric acid 237.10 3.625 [M H] 1 378160 176939 31006 18988 23 N-Ethylglycine 237.10 2.174 [M H] 1 12053298 812656 1665099 97973 24 N-Methylalanine 237.10 2.6 [M H] 1 7789054 2567964 1095566 546229 25 L-alpha-Aminobutyric acid 237.10 1.67 [M H] 1 49720914 3302799 41479142 417895 26 Tartronate semialdehyde 238.05 4.388 [M H] 1 3874452 1242695 286591 81426 27 3-Hydroxybutyric acid 238.08 4.952 [M H] 1 301806216 103894796 5243093 15933411 28 (S)-3-Hydroxyisobutyric acid 238.08 4.035 [M H] 1 102000710 198892671 2677646 6454186 29 (R)-3-Hydroxyisobutyric acid 238.08 4.434 [M H] 1 5691133 8508157 4272373 1139426 30 4-Carboxypyrazole 306.08 2.842 [M H + HAc] 1 4643029 54817 487793 2815 31 (2E,4E)-2,4-Hexadienoic acid 248.10 1.793 [M + H] + 1 440676 353192 407639 230407 32 1-Pyrroline-2-carboxylic acid 247.08 5.843 [M H] 1 510503052 726932 6210566 150549 33 cis-Acetylacrylate 248.07 7.356 [M H] 1 474385 15651520 176040 2408852 34 L-3-Cyanoalanine 248.07 9.49 [M H] 1 1059622 5370099 22694 434557 35 4-Amino-2-methylenebutanoic acid 249.10 2.188 [M H] 1 87350177 56666811 13490729 6935788 36 Levulinic acid 250.08 6.464 [M H] 1 178720991 176955 147448038 11387 37 2-Oxovaleric acid 250.08 9.01 [M H] 1 9445509 945025 2098125 61720 38 2-Methylacetoacetic acid 250.08 7.484 [M H] 1 7951379 8911936 880664 4062180 39 2-Ethylbutanoic acid 250.12 9.023 [M H] 1 12672521 4951315 4077090 786439 40 L-2-Amino-3-oxobutanoic acid 253.09 1.836 [M + H] + 1 73838201 320289 63551844 140244 41 L-Aspartate-semialdehyde 251.08 3.026 [M H] 1 436917309 4211220 36351677 762056 42 N-Methyl-a-aminoisobutyric acid 251.11 1.92 [M H] 1 121887846 61259 11507581 3837 43 5-Aminopentanoic acid 253.13 0.668 [M + H] + 1 3918736 25332320 6415609 20914439 44 4-Hydroxy-2-oxobutanoic acid 252.06 4.852 [M H] 1 27476587 343028 1089113 141601 45 xi-3-Hydroxy-2-oxobutanoic acid 312.08 3.66 [M H + HAc] 1 29764174 77247104 6501749 9010510 46 3-methoxy-3-oxopropanoic acid 312.08 3.087 [M H + HAc] 1 10120248 51191 2417413 3026 47 Fatty acids and conjugates 254.11 2.641 [M + H] + 1 31771483 215413 19499829 173051 48 Fatty acids and conjugates 254.11 2.596 [M + H] + 1 34971729 974769 59722626 785010 49 Fatty acids and conjugates 252.10 6.113 [M H] 1 2254633676 72759156 58768592 8909091 50 Fatty acids and conjugates 252.10 4.507 [M H] 1 358893190 618874 5309460 77632 51 Fatty acids and conjugates 252.10 5.158 [M H] 1 14144656 3494450 3243962 686593 52 Fatty acids and conjugates 252.10 4.92 [M H] 1 49093638 7131098 4340581 2311977 53 L-Allothreonine 253.09 1.592 [M H] 1 23010815 3858495 3438091 570302 54 3-Methylthiopropionic acid 254.06 6.098 [M H] 1 167272734 122956 7300773 12506 55 (S)-3,4-Dihydroxybutyric acid 254.08 2.666 [M H] 1 10373529 2146916 1019102 252088 56 2,4-Dihydroxybutanoic acid 254.08 3.154 [M H] 1 14557796 1523900 2164021 145535 57 4-Deoxyerythronic acid 254.08 2.641 [M H] 1 10369551 2127538 972695 272618 58 4-Deoxythreonic acid 254.08 2.92 [M H] 1 809070 3564964 292135 438578 59 A,b-Dihydroxyisobutyric acid 254.08 5.21 [M H] 1 23965796 10407027 543134 2942333 60 Isonicotinic acid 259.08 2.651 [M + H] + 1 102305718 105725 82584803 114332 61 5-Methylfuran-2-carboxylic acid 262.08 2.396 [M + H] + 1 188653836 8356972 132888236 9881604 62 Imidazoleacetic acid 262.08 1.284 [M + H] + 1 2506979 69477 1328936 14881 63 2-Heptenoic acid 262.12 8.472 [M H] 1 1240023698 284926 112948294 39986 64 1-Pyrroline-4-hydroxy-2-carboxylate 282.12 1.05 [M + NH4] + 1 33383990 392110 33487341 450883 65 Pyrroline hydroxycarboxylic acid 265.09 2.251 [M + H] + 1 384364073 29327068 43796349 48411341 66 (3R,5S)-1-pyrroline-3-hydroxy-5- 263.08 3.694 [M H] 1 3855850257 714755282 carboxylic Acid 223920707 102960754 67 Vigabatrin 265.13 1.761 [M + H] + 1 37085635 38956215 29307615 33732957 68 Methyl hydrogen fumarate 264.06 4.297 [M H] 1 94878164 1018471 6244568 65304 69 2-Hydroxyglutaric acid lactone 264.10 11.6 [M H] 1 25655400 22813723 17304663 6117694 70 Adipate semialdehyde 264.10 10.037 [M H] 1 20240698 4271382 888382 220403 71 4-Acetylbutyrate 264.10 8.366 [M H] 1 1595053 3000545 102771 823663 72 2-Methyl-3-ketovaleric acid 264.10 7.226 [M H] 1 2291298 1838653 171975 443476 73 2-Ketohexanoic acid 264.14 9.335 [M H] 1 1911246 328137 82034 170919 74 3-Oxohexanoic acid 264.14 9.786 [M H] 1 11778657 6661033 265936 1148748 75 2,5-Dioxopentanoate 264.14 9.499 [M H] 1 2791835 590708 847837 306700 76 L-Glutamic gamma-semialdehyde 265.09 3.779 [M H] 1 1534024418 2205185 48867745 1409406 77 3-Hydroxy-L-proline 265.09 1.517 [M H] 1 5432848 14927015 611545 2122487 78 5-Amino-2-oxopentanoic acid 265.09 4.176 [M H] 1 593323230 361362 14981948 32619 79 Beta-Guanidinopropionic acid 267.12 0.553 [M + H] + 1 35379585 10444853 58571246 7341921 80 N-methylvaline 265.13 4.995 [M H] 1 224420851 58040577 18191555 9624895 81 L-Alloisoleucine 265.13 4.809 [M H] 1 29819470 3491846 542615 741341 82 Beta-Leucine 325.15 1.786 [M H + HAc] 1 16061225 223094 2695564 69164 83 (S)-Methylbutanethioic acid 268.09 2.947 [M + H] + 1 78649134 50836 12298198 18687 84 2-Hydroxy-4-oxopentanoic acid 268.09 3.054 [M + H] + 1 12626453 12529 1716166 469 85 2-Acetolactate 266.08 5.355 [M H] 1 498744073 1437981 31880606 311915 86 N-Carbamoylsarcosine 266.11 7.145 [M H] 1 6865075012 3705564 289785550 2438069 87 5-Hydroxyhexanoic acid 266.11 5.321 [M H] 1 16277930 262350 290543 129908 88 2-Ethyl-2-Hydroxybutyric acid 266.11 7.407 [M H] 1 272231157 546354 18134695 366058 89 L-2-Amino-5-hydroxypentanoic acid 267.11 1.416 [M H] 1 1698430 90915 352622 8586 90 2,3-Dihydroxyvaleric acid 268.09 5.419 [M H] 1 1061686411 104712 20065806 24588 91 2-Methylbenzoic acid 270.09 7.638 [M H] 1 2676986841 10257108 122859059 1274107 92 M-toluic Acid 270.09 10.502 [M H] 1 8083724 5066654 500455 742473 93 2-Pyridylacetic acid 271.08 6.732 [M H] 1 515650 54619 130925 36348 94 2-Aminonicotinic acid 274.09 1.901 [M + H] + 1 353328922 2232782 301069464 1384730 95 Trans-urocanate 274.09 1.327 [M + H] + 1 97073030 1579413 166209235 861675 96 Methylimidazoleacetic acid 276.11 1.208 [M + H] + 1 299197480 10759423 289848389 961891 97 Pi-Methylimidazoleacetic acid 276.11 0.581 [M + H] + 1 165875087 1786900 286015596 899069 98 2-Propyl-2,4-pentadienoic acid 274.09 1.884 [M H] 1 115357069 508197 12378801 352127 99 Arecaidine 275.11 8.359 [M H] 1 95267782 24186 6940621 626 100 L-Hypoglycin A 335.14 8.481 [M H + HAc] 1 10331411 32943 597912 1634 101 2-Octenoic acid 276.14 9.801 [M H] 1 9276052 4416796 374158 736373 102 6-Oxopiperidine-2-carboxylic acid 277.09 6.043 [M H] 1 38049394 367162 1546962 53838 103 L-2-Amino-3-methylenehexanoic acid 279.14 2.665 [M + H] + 1 10544073 630369 2839726 1006667 104 3-Hydroxyadipic acid 3,6-lactone 297.12 2.587 [M + NH4] + 1 21236735 78280 21931240 103829 105 4-Hydroxycyclohexylcarboxylic acid 278.11 4.88 [M H] 1 157503606 183149 4691297 40988 106 2-Methylheptanoic acid 278.15 10.759 [M H] 1 20338229 20473965 1864306 2435501 107 Allysine 279.11 1.108 [M H] 1 2807182 314493 696717 20222 108 L-trans-5-Hydroxy-2- 279.11 3.341 [M H] 1 1423756 37975 piperidinecarboxylic acid 268610 2412 109 N-(2-Carboxymethyl)-morpholine 279.11 4.728 [M H] 1 1095546044 407933 71038961 163145 110 N-Propionylalanine 279.11 5.296 [M H] 1 497524956 48065 29571905 8053 111 4-Guanidinobutanoic acid 279.12 2.486 [M H] 1 1581679 88408 213311 5947 112 2-Aminoheptanoate 281.16 1.415 [M + H] + 1 27567023 178365 23590026 42513 113 Monomethyl glutaric acid 282.11 3.182 [M + H] + 1 26040569 389393 6174601 14572 114 (3S)-3,6-Diaminohexanoate 280.14 0.785 [M H] 1 14874828 1136438 3770143 221340 115 (3S,5S)-3,5-Diaminohexanoate 282.16 0.271 [M + H] + 1 3320125 17033 5722971 943 116 L-4-Hydroxyglutamate semialdehyde 283.10 1.575 [M + H] + 1 87944654 8731200 75853946 12166455 117 O-Acetylserine 283.10 1.075 [M + H] + 1 157295107 14428675 269629227 11991693 118 (2R,3R,4R)-2-Amino-4-hydroxy-3- 341.15 1.511 [M H + HAc] 1 2434963 12879539 methylpentanoic acid 635237 3090466 119 3-phenylprop-2-enoic acid 284.10 3.829 [M + H] + 1 356811 69236 97593 13797 120 trans-Cinnamic acid 282.09 8.72 [M H] 1 674959 229561 103340 287646 121 Penicillamine 283.09 4.195 [M H] 1 150845352 7917189 705082 696903 122 (\xc2\xb1)-2-Hydroxy-4- 284.07 6.022 [M H] 1 251478688 101178 (methylthio)butanoic acid 25531414 33234 123 alpha-Fluoro-beta-ureidopropionic acid 286.10 2.635 [M + H] + 1 114904443 41101 68266446 24897 124 2-Deoxypentonic acid 286.10 2.664 [M + H] + 1 43720321 112407 29766910 13529 125 2-Deoxyribonic acid 286.10 2.589 [M + H] + 1 63405399 19779 88251992 4806 126 3-hydroxy-2,2- 284.09 2.71 [M H] 1 14187916 298762 bis(hydroxymethyl)propanoic acid 2142313 101073 127 3,4-Dimethylbenzoic acid 284.10 8.475 [M H] 1 1373693518 4377638 133937468 830103 128 4-Ethylbenzoic acid 285.10 4.733 [M H] 1 391105 61527 6989 46564 129 3-Cresotinic acid 286.08 6.152 [M H] 1 76223888 214577 2500612 139919 130 4-Hydroxy-3-methylbenzoic acid 286.08 5.692 [M H] 1 117011496 1211709 1446748 282448 131 2-Methoxybenzoic acid 286.08 6.66 [M H] 1 21130041 603276 921451 485961 132 L-2-Amino-3-(1-pyrazolyl)propanoic 289.11 0.634 [M H] 1 17310723 15429847 acid 2509618 3088778 133 2-Nonenoic acid 290.15 11.431 [M H] 1 938653 16719871 192209 3136814 134 Tranexamic Acid 292.17 11.374 [M H] 1 17643544 33307648 2945163 1609740 135 Medicanine 295.14 3.02 [M + H] + 1 114383667 27010 2429148 24846 136 Mono-methyl-adipate 294.11 4.03 [M H] 1 164606166 345531 10744545 28815 137 2-Indolecarboxylic acid 297.12 2.517 [M + H] + 1 46182768 731536 3188500 280725 138 trans-S-(1-Propenyl)-L-cysteine 295.10 4.136 [M H] 1 139677933 864747 6290835 119490 139 Glutamic acid gamma-methyl ester 295.10 4.243 [M H] 1 136680935 23031084 6256752 1842721 140 hydroxybutyrylglycine 295.10 2.966 [M H] 1 11551708 215419 2218830 45852 141 Acetylhomoserine 295.10 3.436 [M H] 1 15606445 374548 1140507 73028 142 4-Hydroxycinnamic acid 298.08 7.291 [M H] 1 405911 1277594 6763 433397 143 Enol-phenylpyruvate 298.08 9.173 [M H] 1 1357110 69868 607861 80016 144 2-Phenylbutyric acid 300.13 4.103 [M + H] + 1 39695 17431 944 1723 145 2-methyl-3-phenylpropanoic acid 298.12 9.068 [M H] 1 461003 191382 90219 197450 146 Ethiin 299.08 1.487 [M H] 1 15453907 1797654 4516907 326811 147 3-Pyridinebutanoic acid 299.11 5.575 [M H] 1 247479044 63752454 29561791 14251447 148 xe2\x80\x8bD-\xe2\x80\x8bXylonic 300.08 3.361 [M H] 1 2756231 49636 acid 298648 3297 149 \xe2\x80\x8bL-\xe2\x80\x8bLyxonic acid 300.08 1.772 [M H] 1 104447225 1779717 15622140 282898 150 Arabinonic acid 300.08 2.748 [M H] 1 15623924 104679 3130934 5285 151 3-(3-Hydroxyphenyl)propanoic acid 302.11 3.318 [M + H] + 1 118424972 39503 28282143 3265 152 L-3-Phenyllactic acid 302.11 3.093 [M + H] + 1 264477683 129535 5890811 5343 153 4-Methoxyphenylacetic acid 300.10 6.158 [M H] 1 736422947 241335 9470657 318345 154 Desaminotyrosine 300.10 6.424 [M H] 1 169566041 115990 1708402 7464 155 3-(2-Hydroxyphenyl)propanoic acid 300.10 6.667 [M H] 1 1048006863 96793 65341318 24317 156 4-Hydroxyphenyl-2-propionic acid 300.10 7.58 [M H] 1 454523791 1456452 7092875 255258 157 2,4,7-Decatrienoic acid 300.14 10.717 [M H] 1 2917876 23001139 128098 1659257 158 3-Methoxyanthranilate 303.11 5.247 [M + H] + 1 602388 38221145 520287 5162239 159 3,5-Dihydroxyphenylacetic acid 302.08 4.991 [M H] 1 52800191 115985 718656 31839 160 3-hydroxy-5-methoxybenzoic acid 302.08 4.622 [M H] 1 36256751 240292 2482980 137457 161 2-hydroxy-4-methoxybenzoic acid 302.08 8.429 [M H] 1 22207132 86833441 2765128 17088956 162 cis-4-Decenoic acid 304.17 11.425 [M H] 1 2370592 21248227 152654 574540 163 9-Oxo-nonanoic acid 308.16 4.817 [M + H] + 1 9520814 33059 3588394 14513 164 4-Methylnonanoic acid 306.18 11.823 [M H] 1 12847373 14568161 907376 1428350 165 Gly-Norvaline 310.15 1.318 [M + H] + 1 23644773 167234 16247984 23136 166 Argininic acid 311.15 0.344 [M + H] + 1 4320252 281839 6922250 320176 167 2-Keto-3-deoxy-D-gluconic acid 312.08 3.655 [M H] 1 29764174 77247104 6501749 9010510 168 4-Methoxycinnamic acid 312.10 10.662 [M H] 1 32116255 232400 2003232 98866 169 3-(4-Methylphenyl)oxiranecarboxylic 313.09 11.622 [M H] 1 2827472 16637152 acid 199823 1965749 170 (Z)-10-Hydroxy-8-decene-4,6-diynoic 313.09 6.476 [M H] 1 30002271 10315091 acid 138304 1734816 171 3-Deoxyarabinohexonic acid 316.11 2.405 [M + H] + 1 351310931 6215669 231028882 2120326 172 2-Methyl-3-hydroxy-5-formylpyridine-4- 315.07 8.2 [M H] 1 101162767 26850547 carboxylate 9157274 1483653 173 3,4-Dimethoxybenzoic acid 316.09 5.064 [M H] 1 186522028 120696 1420536 33735 174 3,5-dimethoxybenzoic acid 316.09 5.412 [M H] 1 133874508 2104614 841918 674483 175 2-(3-hydroxy-5-methoxyphenyl)acetic 316.09 5.974 [M H] 1 30487593 45370 acid 417392 21447 176 2-hydroxy-2-(3-methoxyphenyl)acetic 316.09 5.809 [M H] 1 6557547 112869 acid 2121983 99540 177 2-hydroxy-2-(4-methoxyphenyl)acetic 316.09 5.491 [M H] 1 96542022 110346 acid 65077753 87059 178 3-(sulfooxy)butanoic acid 318.04 1.911 [M H] 1 12219282 61577 4023568 3857 179 4-O-Methylgallic acid 318.07 7.051 [M H] 1 3039537 38872274 36750 5252002 180 (Z)-3-Methyl-4-decenoic acid 318.18 11.741 [M H] 1 2177468 4713008 206122 667305 181 2-Hepteneoylglycine 319.14 6.624 [M H] 1 107551942 54158 1217197 3408 182 Pyroglutamylglycine 320.10 3.259 [M H] 1 27875922 16318665 3809371 3069977 183 4,6-Dimethylnonanoic acid 320.20 12.19 [M H] 1 8577731 2548002 885997 2169931 184 N-Heptanoylglycine 323.17 3.735 [M + H] + 1 7592240 10424 194459 745 185 8-Amino-7-oxononanoic acid 323.17 3.75 [M + H] + 1 21001484 10221 153920 650 186 Glycyl-Isoleucine 322.15 3.35 [M H] 1 5635341 52481 1421294 30081 187 5-Hydroxyindoleacetic acid 325.09 6.749 [M H] 1 80976425 47719 4891662 3762 188 4-Anilino-4-oxobutanoic acid 327.11 6.873 [M H] 1 6392717 235533 140654 172660 189 3-Dehydro-L-gulonate 328.08 1.927 [M H] 1 19632441 203936 1938316 8229 190 Monoethyl phthalate 328.09 7.029 [M H] 1 14467813 44235 356071 2248 191 5-(4-hydroxyphenyl)pentanoic acid 328.13 8.105 [M H] 1 183963030 40100 15285663 3044 192 Leucodopachrome 331.10 3.228 [M + H] + 1 526698 19969 420159 961 193 Dopaquinone 329.09 7.028 [M H] 1 1725391 94224 581361 80698 194 L-Dopaquinone 329.09 5.186 [M H] 1 1919681 117223 65458 123659 195 Metyrosine 329.09 5.53 [M H] 1 1512896 55444 881048 30166 196 Gulonic acid 330.09 4.457 [M H] 1 1760105861 8090776 129866114 614995 197 Gluconic acid 330.09 1.694 [M H] 1 141707008 13400913 20621354 3618568 198 (S)-2-(4-Methoxyphenoxy)propanoic 332.12 3.182 [M + H] + 1 91293128 448672 acid 2062863 16791 199 2-(3,5-dimethoxyphenyl)acetic acid 390.13 6.593 [M H + HAc] 1 88547292 54152 2053671 4997 200 (S)-Batatic acid 330.11 6.246 [M H] 1 231228281 57698 5392957 4298 201 L-alpha-Amino-1H-pyrrole-1-hexanoic 332.17 2.958 [M + H] + 1 19478437 17172 acid 2667138 1147 202 2-methyl-4-(sulfooxy)butanoic acid 332.06 4.804 [M H] 1 83155581 39828 3838889 1554 203 5-(sulfooxy)pentanoic acid 332.06 4.37 [M H] 1 36907998 67315 1866491 1757 204 trans-Dodec-2-enoic acid 332.20 12.151 [M H] 1 3414780 3556575 701029 3855169 205 2-octenoylglycine 335.17 3.801 [M + H] + 1 3815752 13985 56993 831 206 3-octenoylglycine 335.17 3.763 [M + H] + 1 3247986 16512 95378 1154 207 N-Acetylaminooctanoic acid 335.17 8.436 [M H] 1 254623752 124045 38033276 7782 208 N-(5-Methyl-3-oxohexyl)alanine 335.17 8.352 [M H] 1 32078566 31831 869294 3475 209 R-2-Hydroxy-3-methylbutanoic acid 3- 336.16 6.468 [M H] 1 8555654 117204 Methylbutanoyl 847309 11661 210 N-Acetylisoputreanine 338.18 3.003 [M + H] + 1 14410756 12787 680847 1274 211 D-Tryptophan 338.13 5.705 [M H] 1 22623825 17866954 3536393 3968802 212 Zeanic acid 339.07 7.299 [M H] 1 4030088 92055 355673 40593 213 3-Indolehydracrylic acid 339.11 7.3 [M H] 1 46977371 2360537 1342344 426453 214 5-Methoxyindoleacetate 339.11 6.338 [M H] 1 32803368 31758 1490625 1898 215 N-Propionylmethionine 341.13 3.291 [M + H] + 1 3791811 11764 435019 981 216 2-Amino-4-hydroxy-6- 343.10 2.643 [M + H] + 1 13643172 73043 pteridinecarboxylic acid 11605388 89828 217 4-(2-Aminophenyl)-2,4-dioxobutanoic 343.10 3.032 [M + H] + 1 4926716 14291 acid 185015 2885 218 triazolopropionic acid 341.10 9.072 [M H] 1 2178392 84917 1048846 6258 219 6-Amino-9H-purine-9-propanoic acid 401.13 4.9 [M H + HAc] 1 175959193 256652 1 9078502 19357 220 3-Phenylpropionylglycine 343.14 3.686 [M + H] + 1 47607214 36444 1077980 14447 221 Dihydrolipoate 402.12 5.421 [M H + HAc] 1 622208098 38010336 38919379 10167513 222 3-(3,4-Dimethoxyphenyl)-2-propenoic 344.12 2.918 [M + H] + 1 17638660 123198 acid 859455 156293 223 3,4-Dihydroxyphenylvaleric acid 346.14 3.305 [M + H] + 1 25948519 13913 2294846 1062 224 2-Methoxy-3-(4- 344.13 6.59 [M H] 1 205245407 52629 methoxyphenyl)propanoic acid 2715599 4856 225 Cucurbic acid 348.19 4.795 [M + H] + 1 15736700 178444 4166507 41304 226 Dihydrojasmonic acid 348.19 4.732 [M + H] + 1 18681975 261157 17465330 57845 227 3-hydroxy-2-methyl-2- 348.05 1.76 [M H] 1 112459094 110962 [(sulfooxy)methyl]propanoic acid 15500812 5421 228 9-Methyldodecanoic acid 348.23 12.844 [M H] 1 277094 2426122 225202 1251444 229 2-hydroxyoct-2-enoylglycine 349.15 10.059 [M H] 1 32712097 95923 966406 8313 230 xi-5-Hydroxydodecanoic acid 350.21 11.513 [M H] 1 9600303 501983 233980 83437 231 Leucyl-Serine 352.16 5.02 [M H] 1 35981421 89528 1015854 24768 232 (2Z)-2-(phenylmethylidene)heptanoic 352.16 5.233 [M H] 1 26862526 69676 acid 899769 29103 233 Glycyl-Phenylalanine 358.15 3.029 [M + H] + 1 6341464 9407 2542718 1899 234 4-(2-Amino-3-hydroxyphenyl)-2,4- 357.08 3.61 [M H] 1 19369899 127600 dioxobutanoic acid 1809789 17641 235 Salsolinol 1-carboxylate 357.12 5.318 [M H] 1 160824385 77612 5190562 63646 236 2-hydroxyphenylpropionylglycine 357.12 9.578 [M H] 1 36220356 72015 8157783 29263 237 Goshuyic acid 358.21 12.391 [M H] 1 4858876 5590352 242757 3093250 238 3-(3,4-dihydroxy-5- 360.08 1.919 [M H] 1 440578 12277 methoxyphenyl)oxirane-2-carboxylic 173758 1054 acid 239 2,3,4,5,6,7-Hexahydroxyheptanoic acid 360.10 4.269 [M H] 1 371551509 70261443 26033213 7432650 240 5-(3,4,5-trihydroxyphenyl)pentanoic acid 362.13 1.317 [M + H] + 1 281456 18467 413594 4098 241 5-(3,5-dihydroxyphenyl)-4- 362.13 2.919 [M + H] + 1 106945309 124907 hydroxypentanoic acid 3957680 158460 242 b4-Hydroxy-5-(3,5-dihydroxyphenyl)- 360.12 5.279 [M H] 1 287090966 26714 valeric acid 16864207 629 243 b4-Hydroxy-(3,4-dihydroxyphenyl)- 360.12 5.514 [M H] 1 60249031 30572 valeric acid 1804785 1395 244 5-Tetradecenoic acid 360.23 12.667 [M H] 1 7452323 5224125 1568864 377259 245 5Z-Tetradecenoic acid 360.23 12.829 [M H] 1 496992 3050486 395093 1241632 246 Tsuzuic acid 360.23 12.75 [M H] 1 1284972 5741761 605943 1385457 247 Mevalonic acid-5P 362.08 3.137 [M H] 1 1116119 35967 860148 2409 248 2-hydroxy-3,4,5-trimethoxybenzoic acid 362.10 5.061 [M H] 1 75884655 117575 1386412 32863 249 12-Methyltridecanoic acid 362.24 13.168 [M H] 1 73882862 67741975 24830050 11433835 250 2,6,10-Trimethylundecanoic acid 362.25 13.078 [M H] 1 19542478 407767 6376340 77424 251 Glycyl-Arginine 365.18 8.09 [M H] 1 2358321 36761 81605 1927 252 Salsoline-1-carboxylate 373.15 3.047 [M + H] + 1 6362589 13105 1683811 5140 253 3-(3,4,5-Trimethoxyphenyl)propanoic 374.14 8.104 [M H] 1 48924645 38687 acid 4581942 2937 254 2-Carboxy-4-dodecanolide 376.19 11.704 [M H] 1 9322395 131970 531703 6679 255 13-Methylmyristic acid 376.26 13.381 [M H] 1 223486451 1364882 76004756 279573 256 beta-Alanyl-L-arginine 379.20 9.022 [M H] 1 6000099 100267 441874 4839 257 3-[3-(Sulfooxy)phenyl]propanoic acid 380.06 5.557 [M H] 1 277496418 88361 5803661 18966 258 3-[4-(sulfooxy)phenyl]propanoic acid 380.06 5.146 [M H] 1 155334437 51223 4568783 1624 259 Dihydrowyerone acid 382.15 3.675 [M + H] + 1 18988081 43576 5619119 4298 260 Hydroxynalidixic acid 382.12 9.365 [M H] 1 1644804 3354488 59028 457929 261 2-{[hydroxy(4-methoxy-1-benzofuran-5- 383.10 10.939 [M H] 1 6323472 118488 yl)methylidene]amino}acetic acid 14619 20811 262 Prolyl-Histidine 386.16 5.532 [M H] 1 3446983 54918 205990 2734 263 7Z,10Z-Hexadecadienoic acid 386.25 12.789 [M H] 1 17375911 2254613 6513568 385728 264 Hypogeic acid 388.26 13.372 [M H] 1 2049467 3205581 1062478 4187612 265 3-hydroxy-3-(3,4,5- 390.13 6.59 [M H] 1 88547292 53525 trimethoxyphenyl)propanoic acid 2053671 4939 266 Isopalmitic acid 390.28 14.227 [M H] 1 798566 6387985 908460 970468 267 Homovanillic acid sulfate 396.05 4.572 [M H] 1 49914729 240546 1799733 29755 268 Dihydrocaffeic acid 3-sulfate 396.05 5.158 [M H] 1 9903409 44531 585484 1412 269 Cinoxacin 396.10 5.28 [M H] 1 144214615 28628 17603505 1112 270 Neuraminic acid 401.13 4.405 [M H] 1 895204947 756226 22167360 218967 271 N-(1-Deoxy-1-fructosyl)serine 401.13 4.742 [M H] 1 247932841 205194 4530101 152930 272 3-Deoxy-D-glycero-D-galacto-2- 404.13 2.649 [M + H] + 1 284073 14630952 nonulosonic acid 225961 12664054 273 9E-Heptadecenoic acid 402.28 13.561 [M H] 1 6881694 7020470 2783189 639263 274 Cyclohexaneundecanoic acid 402.28 13.66 [M H] 1 11737860 6261104 4834761 07290 275 (S)-14-Methylhexadecanoic acid 404.29 14.132 [M H] 1 1727447 5804610 777515 1960928 276 16-Hydroxy hexadecanoic acid 406.27 13.431 [M H] 1 41800724 6689669 17205726 353350 277 (R)-3-Hydroxy-hexadecanoic acid 406.27 12.08 [M H] 1 143844245 7884362 16926271 2287029 278 3-Hydroxyhexadecanoic acid 406.27 13.173 [M H] 1 10222183 2508112 3406065 241374 279 5-Hydroxyhexadecanoic acid 406.27 11.916 [M H] 1 2615889 12988902 214887 3274590 280 7-Hydroxyhexadecanoic acid 406.27 12.695 [M H] 1 197513 5452616 66936 1190164 281 8-Hydroxyhexadecanoic acid 406.27 11.847 [M H] 1 2132446 4991423 223916 1237979 282 3,4,5-trihydroxy-6-[(oxolan-2- 414.15 4.637 [M + H] + 1 19217658 90521 yl)methoxy]oxane-2-carboxylic acid 4411895 6965 283 3,4,5-trihydroxy-6-[(3- 412.14 9.422 [M H] 1 19450281 58025 methylbutanoyl)oxy]oxane-2-carboxylic 943617 2795 acid 284 Calendic acid 414.27 8.649 [M + H] + 1 102647771 5100486 27477508 488517 285 Punicic acid 414.27 8.461 [M + H] + 1 48950252 2630668 19169432 1109662 286 Linolenelaidic acid 414.27 7.48 [M + H] + 1 3003039 27593590 923342 4955418 287 3,4,5-trihydroxy-6-[(3- 414.12 8.168 [M H] 1 706597767 136155 hydroxybutanoyl)oxy]oxane-2- 24989472 54460 carboxylic acid 288 Tyrosyl-Valine 414.18 4.492 [M H] 1 1247190 24904 658172 4829 289 Bovinic acid 416.29 9.488 [M + H] + 1 562037780 2307000 83001976 798698 290 (9E,11E)-Octadecadienoic acid 416.29 9.822 [M + H] + 1 174914828 3328836 21737726 4605771 291 Dihomolinoleic acid 416.29 9.712 [M + H] + 1 488647177 1620981 80759240 160741 292 5-Octadecynoic acid 416.29 11.458 [M + H] + 1 5607810 57252158 3308500 8867798 293 6Z,9Z-octadecadienoic acid 416.29 13.896 [M + H] + 1 67451068 665948 30079106 344192 294 Octadecadienoate 416.29 9.615 [M + H] + 1 259619642 1555275 130161609 347600 295 (10E,12Z)-Octadecadienoic acid 414.28 13.498 [M H] 1 57038511 33771010 23912024 3873450 296 Mangiferic acid 414.28 13.414 [M H] 1 676271339 824087609 266834677 110539718 297 Elaidic acid 416.29 13.89 [M H] 1 345423297 589882789 155279070 97391985 298 Vaccenic acid 416.29 13.978 [M H] 1 123869005 27280370 54460008 5217242 299 16-Methylheptadecanoic acid 418.31 14.424 [M H] 1 10439852 128642288 3930146 34500866 300 xanthurenic acid 8-O-sulfate 419.03 10.048 [M H] 1 517087 205207 514753 88643 301 12-hydroxyheptadecanoic acid 420.29 12.39 [M H] 1 20657364 171209 2632544 72238 302 Orotidine 422.10 3.158 [M H] 1 8172693 30250 6969485 1926 303 Porric acid 422.10 5.508 [M H] 1 36755714 67250 3624668 15454 304 3,5-Dihydroxyphenylvaleric acid sulfate 424.08 5.899 [M H] 1 72314336 47565 3330294 1244 305 3,4,5-trihydroxy-6-[(4-hydroxy-3- 445.16 3.859 [M + NH4] + 1 2047231 96163 methylbut-2-enoyl)oxy]oxane-2- 27120 3734 carboxylic acid 306 4-Hydroxyproline galactoside 487.17 5.655 [M H + HAc] 1 131390623 426038 5723075 109801 307 17-Hydroxylinolenic acid 428.26 11.762 [M H] 1 2567010 5148556 1301141 134212 308 (Z)-13-Oxo-9-octadecenoic acid 430.27 12.372 [M H] 1 4527380 8539429 934039 1171168 309 Avenoleic acid 430.27 12.1 [M H] 1 29642015 92203928 2317532 33278090 310 12-Hydroxy-8,10-octadecadienoic acid 430.27 12.67 [M H] 1 916569 6382616 1164755 1198719 311 (10E,12Z)-9-HODE 430.27 12.474 [M H] 1 5124568 11353262 898704 4357069 312 13-HODE 430.27 12.167 [M H] 1 26291952 77040779 1780901 12269888 313 alpha-Dimorphecolic acid 430.27 11.929 [M H] 1 6069564 21720795 2188196 3055423 314 9,10-Epoxyoctadecenoic acid 430.27 11.858 [M H] 1 4797819 11283123 1032732 2319925 315 12,13-EpOME 430.27 12.875 [M H] 1 2267111 4829389 2281884 847226 316 Phenethylamine glucuronide 430.27 12.257 [M H] 1 9251186 32181745 315903 41126080 317 3-Oxooctadecanoic acid 432.29 12.823 [M H] 1 1572760 2572358 174555 511016 318 9-Oxooctadecanoic acid 432.29 12.469 [M H] 1 20085576 11604607 4030884 1765007 319 10-Oxooctadecanoic acid 432.29 12.339 [M H] 1 510669565 9396774 89374880 8198094 320 11-Oxooctadecanoic acid 432.29 12.277 [M H] 1 122641516 22288090 17057886 15318462 321 5-Hexyltetrahydro-2-furanoctanoic acid 432.29 12.883 [M H] 1 399730 1123125 64764 334858 322 (R)-3-Hydroxy-Octadecanoic acid 434.30 12.56 [M H] 1 1284577242 7545156 305296771 4668860 323 xi-10-Hydroxyoctadecanoic acid 434.30 12.695 [M H] 1 27938700 2261349 5715126 91408 324 Pyrogallol-2-O-glucuronide 436.10 11.189 [M H] 1 539805 3253620 160230 562979 325 6-(2,4-dihydroxyphenoxy)-3,4,5- 436.10 11.349 [M H] 1 2213769 21151840 trihydroxyoxane-2-carboxylic acid 719244 2364104 326 6-(2,5-dihydroxyphenoxy)-3,4,5- 436.10 11.63 [M H] 1 2068500 22519723 trihydroxyoxane-2-carboxylic acid 373176 1015590 327 6-(3,4-dihydroxyphenoxy)-3,4,5- 436.10 11.46 [M H] 1 3247448 29391873 trihydroxyoxane-2-carboxylic acid 664244 2501473 328 Porric acid A 436.11 5.843 [M H] 1 92312578 35725 3795837 431 329 Cis-8,11,14,17-Eicosatetraenoic acid 440.29 11.266 [M + H] + 1 1733568 53280633 489736 41031354 330 2-(3,4-Dihydroxybenzoyloxy)-4,6- 440.07 13.6 [M H] 1 356155 82829 dihydroxybenzoate 250378 2856 331 5-(3,4-dihydroxyphenyl)-4- 440.08 4.794 [M H] 1 176423351 34249 (sulfooxy)pentanoic acid 2727594 1355 332 3,4-dihydroxy-2-(4-hydroxy-3,7- 440.18 0.955 [M H] 1 21206815 535907 dimethylocta-2,6-dien-1-yl)benzoic acid 18068363 30832 333 3,4-dihydroxy-2-[(2Z)-4-hydroxy-3-(4- 440.18 0.91 [M H] 1 34747475 1427681 methylpent-3-en-1-yl)but-2-en-1- 7183172 727074 yl]benzoic acid 334 b5-Carboxy-gamma-chromanol 442.23 5.534 [M + H] + 1 39410744 232039 770524 10693 335 5,8,11-Eicosatrienoic acid 440.29 13.577 [M H] 1 10228924 16089071 5254138 2598430 336 Corchorifatty acid A 442.23 3.328 [M H] 1 39191074 16582274 8328846 2020810 337 Eicosadienoic acid 442.31 13.969 [M H] 1 1632999 5330577 1028883 825514 338 3,4,5-trihydroxy-6-{[3-hydroxy-2- 446.14 3.287 [M + H] + 1 14686973 144711 (hydroxymethyl)-2- 385197 36494 methylpropanoyl]oxy}oxane-2- carboxylic acid 339 (R)-2-Hydroxysterculic acid 444.29 12.867 [M H] 1 2656442 3269807 1286306 212336 340 8(R)-Hydroperoxylinoleic acid 446.27 12.188 [M H] 1 12593976 37783556 1999137 11286390 341 (\xc2\xb1)-(E)-13-Hydroxy-10-oxo-11- 446.27 12.051 [M H] 1 2831245 8969604 octadecenoic acid 905725 2221456 342 Beta-D-Glucopyranuronic acid 448.10 12.379 [M H] 1 317677 8922364 165203 674867 343 3,4,5-trihydroxy-6-(3- 448.10 11.648 [M H] 1 422340 10693456 hydroxybenzoyloxy)oxane-2-carboxylic 54500 5702913 acid 344 9,10-Epoxyoctadecanoic acid 448.28 11.641 [M H] 1 87004989 1374409 3973815 184155 345 12,13-DHOME 448.28 12.358 [M H] 1 1026380 2297055 608068 1494177 346 9,10-DHOME 448.28 12.418 [M H] 1 1898753 3003796 312906 1371028 347 (+)-15,16-Dihydroxyoctadecanoic acid 452.31 7.996 [M + H] + 1 182080968 1005144 49497922 477658 348 (9S,10S)-9,10-dihydroxyoctadecanoate 452.31 7.597 [M + H] + 1 33056391 961508 2714072 142134 349 Valproic acid glucuronide 456.20 3.676 [M + H] + 1 32799856 123805 3113321 7280 350 2-Hydroxy-6-tridecylbenzoic acid 454.27 12.179 [M H] 1 371798 10320632 232645 7951616 351 19(S)-HETE 454.27 12.35 [M H] 1 267167 11165932 38896 1027869 352 10-HETE 454.27 12.615 [M H] 1 203539 41648687 48719 2147758 353 13-HETE 454.27 12.428 [M H] 1 437565 15692639 96921 3729459 354 12 Hydroxy arachidonic acid 454.27 12.282 [M H] 1 222454 11264070 74787 2210161 355 12S-hydroxy-5E,8Z,10Z,14Z- 454.27 12.451 [M H] 1 100756 4218983 eicosatetraenoic acid 17609 1545730 356 Lepidimoic acid 458.14 3.472 [M + H] + 1 24066028 19662 5739940 1735 357 3,4-Dimethyl-5-pentyl-2-furannonanoic 456.29 12.047 [M H] 1 23661841 256504 acid 1984650 95569 358 Corchorifatty acid F 463.29 12.897 [M H] 1 8172175 135246 2165706 6453 359 9,12,13-TriHOME 464.28 9.733 [M H] 1 54386843 592064 3628022 27726 360 9,10,13-TriHOME 464.28 9.812 [M H] 1 111662018 1929528 7179078 597563 361 4,8,12,15,19-Docosapentaenoic acid 464.29 13.374 [M H] 1 3022443 2386925 1435794 484525 362 9,10,13-Trihydroxystearic acid 466.29 10.34 [M H] 1 15563703 109258 377280 6956 363 Prostaglandin B2 468.25 11.539 [M H] 1 10813788 184849 1406868 117650 364 3,4,5-trihydroxy-6-{[(2E)-3-(3- 474.12 12.01 [M H] 1 285412 10033224 hydroxyphenyl)prop-2- 36657 561452 enoyl]oxy}oxane-2-carboxylic acid 365 3,4,5-trihydroxy-6-{[(2E)-3-(4- 474.12 11.83 [M H] 1 561451 15328466 hydroxyphenyl)prop-2- 154877 2978586 enoyl]oxy}oxane-2-carboxylic acid 366 3,4,5-trihydroxy-6-{[3-(3- 474.12 12.27 [M H] 1 230347 4180602 hydroxyphenyl)prop-2- 28396 2422104 enoyl]oxy}oxane-2-carboxylic acid 367 3,4,5-trihydroxy-6-[3-(4- 478.18 3.776 [M + H] + 1 6144013 8744 methoxyphenyl)propoxy]oxane-2- 619614 750 carboxylic acid 368 13-Hydroxy-9-methoxy-10-oxo-11- 476.28 12.356 [M H] 1 707597 11725314 octadecenoic acid 99637 9946207 369 2-Hydroxy-6-pentadecylbenzoic acid 482.30 11.881 [M H] 1 12770057 151740 1591691 2572 370 4-Methylumbelliferone glucuronide 486.12 11.739 [M H] 1 749110 27793207 44634 946668 371 Cryptochlorogenic acid 488.13 11.943 [M H] 1 1338251 52406067 89179 6324506 372 5-Caffeoylquinic acid 488.13 11.81 [M H] 1 1001686 47163602 629003 2046485 373 3,4,5-trihydroxy-6-{[3-(3- 488.13 11.33 [M H] 1 764298 65035070 methoxyphenyl)prop-2- 193256 4754398 enoyl]oxy}oxane-2-carboxylic acid 374 3,4,5-trihydroxy-6-{[(2E)-3-(3- 488.13 12.013 [M H] 1 358182 37143083 methoxyphenyl)prop-2- 28055 645807 enoyl]oxy}oxane-2-carboxylic acid 375 3,4,5-trihydroxy-6-{[(2E)-2- 488.13 11.865 [M H] 1 3288892 164304226 (hydroxymethyl)-3-phenylprop-2- 904240 19453671 enoyl]oxy}oxane-2-carboxylic acid 376 3,4,5-trihydroxy-6-{[(2E)-3-(3- 488.13 11.716 [M H] 1 5244846 257754514 hydroxyphenyl)-2-methylprop-2- 1462206 29782660 enoyl]oxy}oxane-2-carboxylic acid 377 3,4,5-trihydroxy-6-{[(2E)-3-(4- 488.13 11.641 [M H] 1 1183413 110937452 hydroxyphenyl)-2-methylprop-2- 344044 11980258 enoyl]oxy}oxane-2-carboxylic acid 378 3,4,5-trihydroxy-6-{[(2E)-3-(2- 488.13 11.572 [M H] 1 552018 52501894 hydroxyphenyl)-2-methylprop-2- 248208 2188003 enoyl]oxy}oxane-2-carboxylic acid 379 3,4,5-trihydroxy-6-{[3-(4- 488.13 11.208 [M H] 1 764298 11449342 methoxyphenyl)prop-2- 193256 2788799 enoyl]oxy}oxane-2-carboxylic acid 380 Sulindac 490.12 12.5 [M H] 1 248999 1067937 124486 44336 381 3,4,5-trihydroxy-6-[5-hydroxy-2- 492.16 7.476 [M + H] + 1 415073 32153032 methoxy-4-(prop-2-en-1- 50302 1481907 yl)phenoxy]oxane-2-carboxylic acid 382 3,4,5-trihydroxy-6-[5-hydroxy-4- 492.16 6.333 [M + H] + 1 265877 30343221 methoxy-2-(prop-2-en-1- 117576 1013625 yl)phenoxy]oxane-2-carboxylic acid 383 3,4,5-trihydroxy-6-{[3-(3- 492.16 6.393 [M + H] + 1 283221 80304902 methoxyphenyl)propanoyl]oxy}oxane-2- 98823 15929303 carboxylic acid 384 3,4,5-trihydroxy-6-{[2-(4- 490.15 11.461 [M H] 1 231668 2885292 methoxyphenyl)propanoyl]oxy}oxane-2- 24742 641625 carboxylic acid 385 Tetracosahexaenoic acid 492.32 7.847 [M + H] + 1 7220101994 1726046 144799958 120207 386 6,9,12,15,18,21-Tetracosahexaenoic acid 492.32 6.284 [M + H] + 1 171476795 201112 42496040 263735 387 Tetracosahexaenoic acid, n-3 492.32 5.731 [M + H] + 1 275527792 517583 16577519 407316 388 5-hydroxy-2-{[1-hydroxy-3-(4-hydroxy- 493.13 6.238 [M H] 1 83824095 53113 3-methoxyphenyl)prop-2-en-1- 5817825 7086 ylidene]amino}-4-methoxybenzoic acid 389 Rosmarinic acid 494.12 6.676 [M H] 1 69856616 37698 5550646 3154 390 Feruloyl C1-glucuronide 504.13 11.828 [M H] 1 1150529 4030880 12608 859188 391 Isoferuloyl C1-glucuronide 504.13 10.569 [M H] 1 761604 11722683 15570 1247781 392 3,4,5-trihydroxy-6-{[3-(4-hydroxy-3- 504.13 10.791 [M H] 1 366531 4752867 methoxyphenyl)prop-2- 48454 340214 enoyl]oxy}oxane-2-carboxylic acid 393 3,4,5-trihydroxy-6-{[3-(3-hydroxy-5- 504.13 11.361 [M H] 1 978673 19126186 methoxyphenyl)prop-2- 64639 674346 enoyl]oxy}oxane-2-carboxylic acid 394 3,4,5-trihydroxy-6-{[(2E)-3-(4-hydroxy- 504.13 11.203 [M H] 1 541613 10096751 3-methoxyphenyl)prop-2- 25396 2230534 enoyl]oxy}oxane-2-carboxylic acid 395 3,4,5-trihydroxy-6-{[3-(2-hydroxy-4- 504.13 10.956 [M H] 1 403501 9765280 methoxyphenyl)prop-2- 13037 2718087 enoyl]oxy}oxane-2-carboxylic acid 396 3,4,5-trihydroxy-6-{[3-(4- 504.13 10.69 [M H] 1 451786 5949088 methoxyphenyl)-2- 31638 1085216 oxopropanoyl]oxy}oxane-2-carboxylic acid 397 (3R)-3-(tetradecanoyloxy)-4- 507.35 10.39 [M + H] + 1 188234605 871188 (trimethylazaniumyl)butanoate 73194844 199932 398 Ethyl gallate 3-glucuronide 508.12 8.985 [M H] 1 105752943 163314 45846541 7882 399 2-(10-Heptadecenyl)-6-hydroxybenzoic 508.32 13.021 [M H] 1 12794494 97035 acid 4306336 3888 400 12b-Hydroxy-5b-cholanoic acid 510.33 12.428 [M H] 1 641446664 949703 189084164 735439 401 Allolithocholic acid 510.33 12.025 [M H] 1 115231454 359177 38319237 362857 402 7a-Hydroxy-5b-cholanic acid 510.33 11.916 [M H] 1 1156055 250890 338435 247541 403 6-[(4,7-dihydroxy-2,2-dimethyl-3,4- 520.16 12.006 [M H] 1 1870162 11915130 dihydro-2H-1-benzopyran-5-yl)oxy]- 475219 2335910 3,4,5-trihydroxyoxane-2-carboxylic acid 404 Treprostinil 524.28 12.31 [M H] 1 980525 8520699 523501 451539 405 bD8-Merulinic acid A 524.31 11.366 [M H] 1 541158745 449488 374961606 279265 406 7-Hydroxy-3-oxocholanoic acid 524.31 11.592 [M H] 1 5235781 169193 3448206 8232 407 11-Hydroperoxy-H4-neuroprostane 526.26 11.574 [M H] 1 331036 100458 85161 838 408 3a,12b-Dihydroxy-5b-cholanoic acid 526.33 11.944 [M H] 1 1812605216 4679585 192418223 378889 409 3b,12a-Dihydroxy-5a-cholanoic acid 526.33 10.534 [M H] 1 679106438 388461 40615214 78335 410 3b,7a-Dihydroxy-5b-cholanoic acid 526.33 11.778 [M H] 1 548961777 2777820 30873343 251814 411 3a,7a-Dihydroxycholanoic acid 526.33 10.902 [M H] 1 660581393 322672 34926669 217922 412 Allodeoxycholic acid 526.33 11.604 [M H] 1 44284334 506183 852858 327197 413 Allochenodeoxycholic acid 526.33 11.871 [M H] 1 29068727 186069 3682524 5434 414 Murocholic acid 526.33 11.249 [M H] 1 148446058 287273 9847095 26392 415 7a,12b-dihydroxy-5b-Cholan-24-oic acid 526.33 11.33 [M H] 1 932019319 436568 348382725 286077 416 3beta,12beta-Dihydroxy-5beta-cholanoic 526.33 12.199 [M H] 1 232158793 476448 acid 10792209 7844 417 Sinapinic acid-O-glucuronide isomer 534.14 11.937 [M H] 1 1217503 3553889 81133 730667 418 3,4,5-trihydroxy-6-{[6-(3-hydroxyprop- 534.14 11.325 [M H] 1 1832391 142882837 1-en-1-yl)-5-methoxy-2H-1,3- 1084795 7978855 benzodioxol-4-yl]oxy}oxane-2- carboxylic acid 419 3,4,5-trihydroxy-6-[2-hydroxy-3,4- 534.14 11.57 [M H] 1 634692 53948195 dimethoxy-5-(3-oxoprop-1-en-1- 486133 4992807 yl)phenoxy]oxane-2-carboxylic acid 420 b5-(3,4,5-Trihydroxyphenyl)-gamma- 534.14 11.654 [M H] 1 387193 51877403 valerolactone-3-O-glucuronide 106043 14353027 421 b5-(3,4,5-Trihydroxyphenyl)-gamma- 536.19 4.204 [M + H] + 1 25890202 146487 valerolactone-4-O-glucuronide 1901296 4740 422 b4-Hydroxy-5-(3,4-dihydroxyphenyl)- 536.15 11.452 [M H] 1 311804 39471028 valeric acid-O-glucuronide 56347 2427874 423 6-{[5-(3,4-dihydroxyphenyl)-4- 536.15 11.878 [M H] 1 193855 18955563 hydroxypentanoyl]oxy}-3,4,5- 22678 1049628 trihydroxyoxane-2-carboxylic acid 424 3,4,5-trihydroxy-6-[2-hydroxy-3,4- 536.15 11.51 [M H] 1 343919 35256359 dimethoxy-5-(3- 116260 3190656 oxopropyl)phenoxy]oxane-2-carboxylic acid 425 7-Ketodeoxycholic acid 542.32 4.197 [M + H] + 1 157854029 156172 9462565 5053 426 3,7-Dihydroxy-12-oxocholanoic acid 542.32 3.838 [M + H] + 1 9016731 10773 878392 1084 427 1b,3a,12a-Trihydroxy-5b-cholanoic acid 542.32 10.137 [M H] 1 167611063 114235 4327022 4658 428 1b,3a,7a-Trihydroxy-5b-cholanoic acid 542.32 11.148 [M H] 1 544907160 3155128 4565684 56662 429 3alpha,7alpha,12beta-Trihydroxy-5beta- 542.32 9.526 [M H] 1 1241848325 64094 cholanoic acid 21099752 2673 430 3a,4b,7a-Trihydroxy-5b-cholanoic acid 542.32 10.707 [M H] 1 3495175556 674030 579880 577596 431 3a,4b,12a-Trihydroxy-5b-cholanoic acid 542.32 9.706 [M H] 1 195680988 224977 12-Ketodeoxycholic acid 10657635 9852 432 1,3,12-Trihydroxycholan-24-oic acid 542.32 8.65 [M H] 1 925432007 88584 59000243 12829 433 3b,7a,12a-Trihydroxy-5a-Cholanoic acid 542.32 9.97 [M H] 1 27588201 429960 4722587 32149 434 3a,7b,12b-Trihydroxy-5b-cholanoic acid 542.32 8.09 [M H] 1 35540636 38071 3548529 1995 435 3b,7b,12a-Trihydroxy-5b-cholanoic acid 542.32 8.82 [M H] 1 25513288 75397 9016311 3160 436 2b,3a,7a-Trihydroxy-5b-cholanoic acid 542.32 8.973 [M H] 1 21080013 118675 14979217 8489 437 Trovafloxacin 552.16 5.011 [M + H] + 1 25987908 16523986 5996264 1033800 438 3,4,5-trihydroxy-6-{4-hydroxy-2-[(2E)- 550.14 10.286 [M H] 1 48373249 18790255 3-phenylprop-2-enoyl]phenoxy}oxane-2- 12447522 2443497 carboxylic acid 439 3,4,5-trihydroxy-6-[4-hydroxy-3-(3- 550.14 10.386 [M H] 1 44046888 14346146 phenylprop-2-enoyl)phenoxy]oxane-2- 12250562 2123539 carboxylic acid 440 7,14-dihydroxy-6-methoxy-4,12- 562.14 11.722 [M H] 1 199302 8211896 dimethyl-5-(3-methylbut-2-enoyl)-10- 25981 372731 oxo-2,9- dioxatricyclo[9.4.0.0\xc2\xb3,\xe2\x81\ xb8]pentadeca-1(11),3(8),4,6,12,14- hexaene-15-carboxylic acid 441 6-[2-(benzoyloxy)-5-(prop-2-en-1- 564.16 10.648 [M H] 1 240806 196687 yl)phenoxy]-3,4,5-trihydroxyoxane-2- 7836 120183 carboxylic acid 442 3,4,5-trihydroxy-6-{2-[(1E)-3-(4- 564.16 11.847 [M H] 1 812681 54894976 methoxyphenyl)-3-oxoprop-1-en-1- 311959 17140906 yl]phenoxy}oxane-2-carboxylic acid 443 3,4,5-trihydroxy-6-{5-methoxy-2-[(2E)- 564.16 11.781 [M H] 1 879920 46776254 3-phenylprop-2-enoyl]phenoxy}oxane-2- 197511 7914696 carboxylic acid 444 3,4,5-trihydroxy-6-{4-[(1E)-3-(4- 564.16 11.532 [M H] 1 1396695 54509330 methoxyphenyl)-3-oxoprop-1-en-1- 92231 19840841 yl]phenoxy}oxane-2-carboxylic acid 445 3,4,5-trihydroxy-6-{3-[(1E)-3-(4- 564.16 11.953 [M H] 1 1383091 10848077 methoxyphenyl)-3-oxoprop-1-en-1- 92167 3236227 yl]phenoxy}oxane-2-carboxylic acid 446 Ketoprofen glucuronide 564.16 11.213 [M H] 1 2544589 4777722 1554692 1434362 447 3,4,5-trihydroxy-6-{[8-methoxy-6-(3- 570.17 4.615 [M H] 1 228973368 600200 methylbut-2-en-1-yl)-2-oxo-2H- 15107157 74242 chromen-7-yl]oxy}oxane-2-carboxylic acid 448 simvastatin hydroxy acid 570.32 11.389 [M H] 1 34301301 166890 5036658 11744 449 Varanic acid 570.35 11.427 [M H] 1 3360794 99464 262300 4778 450 Formononetin 7-glucuronide 580.15 3.987 [M + H] + 1 77327 194116 55313 131885 451 3,4,5-trihydroxy-6-[(8-methoxy-4-oxo-2- 580.15 8.439 [M H] 1 115818670 104279 phenyl-3,4-dihydro-2H-1-benzopyran-7- 14734095 6542 yl)oxy]oxane-2-carboxylic acid 452 N-Stearoyl tyrosine 581.37 12.27 [M H] 1 12635191 120789 2188071 2548 453 3alpha,7alpha,12alpha-trihydroxy-5beta- 584.37 11.903 [M H] 1 14617490 96856 cholestanate 1062188 3422 454 3,4,5-trihydroxy-6-{6-hydroxy-3-[3-(3- 596.15 8.486 [M H] 1 20764642 27169164 hydroxyphenyl)-3-oxoprop-1-en-1-yl]-2- 1225036 5774647 methoxyphenoxy}oxane-2-carboxylic acid 455 (3alpha,20R,24Z)-3-Hydroxy-21- 606.39 9.368 [M + H] + 1 98724208 4768874 oxoeupha-8,24-dien-26-oic acid 33043801 165596 456 23-Hydroxy-3-oxocycloart-24-en-26-oic 604.38 12.257 [M H] 1 7915874 95440 acid 2772630 9300 457 Chenodeoxycholic acid 3-sulfate 606.29 9.184 [M H] 1 108202813 28217 38912268 632 458 Ursodeoxycholic acid 3-sulfate 606.29 10.816 [M H] 1 200146167 350728 64217935 420289 459 Chenodeoxycholic acid sulfate 606.29 9.759 [M H] 1 18347698 95308 4837735 4174 460 b4-O-Methylepicatechin 7-O- 614.16 7.92 [M H] 1 65767168 92340 glucuronide 3359931 17237 461 6-{5-[(E)-2-(2,4- 622.20 10.042 [M H] 1 1486863 171224 dihydroxyphenyl)ethenyl]-3-hydroxy-2- 459795 11362 (3-methylbut-2-en-1-yl)phenoxy}-3,4,5- trihydroxyoxane-2-carboxylic acid 462 7-Sulfocholic acid 622.28 9.981 [M H] 1 21115252 339947 1446294 25419 463 Fexofenadine 635.32 10.714 [M H] 1 4092130 224486 121573 4692 464 3,4,5-trihydroxy-6-({5,6,14-trimethoxy- 640.18 10.132 [M H] 1 83141191 132244 8,17- 16073507 5392 dioxatetracyclo[8.7.0.0\xc2\xb2,\xe2\x81\ xb7.0\xc2\xb9\xc2\xb9,\xc2\xb9\xe2\x81\ xb6]heptadeca-2,4,6,11(16),12,14- hexaen-4-yl}oxy)oxane-2-carboxylic acid 465 4-{7-[(6-carboxy-3,4,5-trihydroxyoxan- 648.12 12.011 [M H] 1 337835 6657664 2-yl)oxy]-3-hydroxy-5-sulfino-3,4- 43390 1417613 dihydro-2H-1-benzopyran-2-yl}-2- hydroxybenzen-1-olate 466 3,3-Diiodothyronine 658.93 7.666 [M H] 1 778652 66407 28870 38205 467 6-{[6-(3,4-dihydroxy-6-methyl-5- 740.19 3.802 [M + H] + 1 2510636 15222 oxooxan-2-yl)-5-hydroxy-2-(3- 93502 1211 methoxyphenyl)-4-oxo-4H-chromen-7- yl]oxy}-3,4,5-trihydroxyoxane-2- carboxylic acid