Liquid-Chromatographic Methods for Carboxylic Acids

Liquid-Chromatographic Methods for Carboxylic Acids: History

Please note this is an old version of this entry, which may differ significantly from the current revision.

Subjects: Chemistry, Analytical

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Carboxyl-bearing low-molecular-weight compounds such as keto acids, fatty acids, and other organic acids are involved in a myriad of metabolic pathways owing to their high polarity and solubility in biological fluids. Various disease areas such as cancer, myeloid leukemia, heart disease, liver disease, and lifestyle diseases (obesity and diabetes) were found to be related to certain metabolic pathways and changes in the concentrations of the compounds involved in those pathways. Therefore, the quantification of such compounds provides useful information pertaining to diagnosis, pathological conditions, and disease mechanisms, spurring the development of numerous analytical methods for this purpose.

fluorescence
mass spectrometry
fatty acids
perfluorinated carboxylic acids
α-keto acids

1. Introduction

Quantification of low-molecular-weight compounds, as exemplified by metabolomics studies, has become increasingly important in the life sciences. Metabolite analysis provides metabolic and biochemical status of particular biological systems and valuable insights into disease development and diagnosis ^[1]^[2]^[3]^[4]^[5]^[6]. There are numerous classes of low-molecular-weight compounds, and they are categorized based on their functional groups, including amine, thiol, and carboxylic groups. Low-molecular-weight carboxylic acids are involved in various metabolic pathways. For example, the tricarboxylic acid (TCA) cycle, which is the principal energy-producing process in cells, involves nine carboxylic acid compounds. Fatty acids are integral components of lipids, and consist of carboxylic acids with long aliphatic chains.

Hence, highly sensitive and selective methods for the determination of biologically important carboxylic acids are required for biological investigations, and, thus far, numerous analytical methods have been developed. For selective determination, solid-phase extraction or solvent extraction pretreatment is commonly performed, followed by separation techniques such as liquid chromatography (LC), gas chromatography (GC), and capillary electrophoresis. The choice of detection method is important for trace amounts of carboxylic acids in biological samples. Ultraviolet absorbance detection is rarely implemented due to the absence of chromophores in carboxylic acids. Fluorescence detection following derivatization and mass spectrometry has the advantage of high sensitivity.

2. Analytical methods for fatty acids in biological samples

Target Compounds

Biological Sample

Sample Treatment

Derivatization Reagent

Separation Mode

Detection Method

LOD

Recovery

Ref.

7 Fatty acids

Human serum

Acid extraction

APF

RPLC

FL: 467/512 nm

0.1–6.4 nM

93–105%

^[7]

3 Fatty acids

Human plasma

Acid extraction

NOEPES

RPLC

FL: 235/366 nm

56 fmol

–

^[8]

6 Fatty acids

Human plasma

Acid extraction

HEC

RPLC

FL: 293/365 nm

38–57 fmol

102–106%

^[9]

6 Fatty acids

Human plasma

Acid extraction

HEC

RPLC

FL: 335/365 nm

45–68 fmol

102–105%

^[10]

5 Fatty acids

Human serum

Acid extraction

DBD-ED

RPLC

FL: 450/560 nm

2.29–4.75 fmol

108–113%

^[11]

8 Fatty acids

Rat plasma

Acid extraction

DBD-ED

RPLC

FL: 450/560 nm

–

^[12]

4 Epoxyeicosatrienoic acids

Bovine endothelial cells

Solid phase extraction

RPLC

FL: 259/395 nm

<2 pg

83–89%

^[13]

25 Fatty acids

Mouse serum

Acid extraction

AMPP

RPLC

MS/MS

50–100 fg (LOQ)

–

^[14]

11 Fatty acids

Mouse serum, bronchial epithelial cells

Solid phase extraction

AMPP

RPLC

MS/MS

200–900 fg (LOQ)

–

^[15]

20 Fatty acids

Breast cancer cells

Solvent extraction

Aminoxy

TMT

RPLC

MS/MS

40 fmol

–

^[16]

8 Fatty acids

Rat plasma

Acid extraction

DBD-PZ-NH₂

RPLC

<0.1 µM

–

^[17]

9 Fatty acids

Rat plasma

Solvent extraction

DAABD-AE

RPLC

6.5–21 fmol

–

^[18]

MePZBD-AE

RPLC

8.8–32 fmol

–

^[18]

APZBD-NHMe

RPLC

35–150 fmol

–

^[18]

56 Fatty acids

Human plasma

Centrifugation

Choline

HILIC

50 ng/mL

–

^[19]

38 Fatty acids, acylcarnitines

Human plasma

Centrifugation

Dansyl-hydrazine

RPLC

MS/MS

76–152 pM

–

^[20]

18 Fatty acids

Human urine

Solid phase extraction

d₀-DMPP,

d₆-DMPP

RPLC

MS/MS

5–15 pM

–

^[21]

60 Fatty acids

Human serum

Acid extraction

DMED, d₄-DMED

RPLC

–

^[22]

6 Fatty acids

Human blood

Acid extraction

None

RPLC

low pg range

–

^[23]

4 Fatty acids

Human serum, plasma

Solvent extraction

None

RPLC

ECD

50 pmol

92–102%

^[24]

6 Fatty acids

Human plasma

Solvent extraction

None

RPLC

ECD

50 pmol

92–102%

^[25]

11 Fatty acids

Human plasma

Solvent extraction

AEMP, NAPP

RPLC

Electrogenerated chemilu minescence

70 fmol

–

^[26]

APF: 6-oxy-(acetyl piperazine)fluorescein, NOEPES: 2-(2-naphoxy)ethyl 2-(piperidino)ethanesulfonate, HEC: 9-(2-hydroxyethyl)-carbazole, DBD-ED: 4-N,N-dimethylaminosulfonyl-7-N-(2-aminoethyl)amino-2,1,3-benzoxadiazole, NT: 2-(2,3-naphthalimino)ethyl trifluoromethanesulfonate, AMPP: N-(4-aminomethylphenyl)pyridinium, AminoxyTMT: aminoxy tandem mass tags, DBD-PZ-NH₂: 7-(N,N-dimethylaminosulfonyl)-4-(aminoethyl)piperazino-2,1,3-benzoxadiazole, DAABD-AE: 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole, MePZBD-AE: [4-(4-N-methyl)piperazinosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole, APZBD-NHMe: [4-(4-N-aminoethyl)piperazinosulfonyl]-7-methylamino-2,1,3-benzoxadiazole, DMPP: 2,4-dimethoxy-6-piperazin-1-yl pyrimidine, DMED: 2-dimethylaminoethylamine, AEMP: 2-(2-aminoethyl)-1-methylpyrrolidine, NAPP: N-(3-aminopropyl)pyrrolidine.

3. Analytical methods for TCA cycle and glycolysis-related compounds in biological samples

Target Compounds	Biological Sample	Sample Treatment	Derivatization Reagent			Separation Mode	Detection Method	LOD	Recovery	Ref.
Fumaric acid	Rat liver, spleen and urine	Centrifugation			None	RPLC	PDA: 215 nm	0.01 µg	89–92%	^[27]
Maleic acid	Rat serum and urine	Centrifugation			None	RPLC	MS/MS	0.2 µg/L	94–111%	^[28]
Methylma-lonic acid	Human plasma	Centrifugation			None	HILIC	MS	0.03 µM	90–93%	^[29]
Lactic acid	Human urine and saliva	Centrifugation			9-CMA	RPLC	UV: 365 nm, FL: 365/410 nm	50 nM	92–106%	^[30]
Oxalic acid	Mouse urine and hepatocyte	Centrifugation			None	Ion exclu sion chrom atography	MS/MS	2 µM	–	^[31]
6 TCA metabolites	Rat urine	Centrifugation			DBD-PZ	RPLC	FL: 450/560 nm	2–15 fmol	80–96%	^[32]
9 Organic acids	Yeast	Centrifugation		None		Ion exclu sion chrom atography	UV: 210 nm	0.6–29.3 mg/L	98–103%	^[33]
32 Organic acids	Human urine	Solvent extraction		None		Ion exclu sion chrom atography	UV: 220 nm	0.002–2.2 mg/L	–	^[34]
13 Organic acids	Mouse urine	Centrifugation		1-Pyrene methylamine		RPLC	FL: 345/375, 345/475 nm	4–22 fmol	–	^[35]
30 Organic acids	Mouse ser um, urine, and tissue	Centrifugation		None		HILIC, Ion pair RPLC	MS/MS	<5 µM	–	^[36]
59 Organic acids	Human melanoma cells	Centrifugation		Phenylh-drazine		Ion pair RPLC	MS	–	–	^[37]
138 Organic acids	Yeast	Centrifugation		None		RPLC	MS/MS	0.001–3.7 µM	–	^[38]
TCA metabolites	Human red blood cell	Centrifugation		None		RPLC	MS	–	–	^[39]

9-CMA: 9-chloromethyl anthracene, DBD-PZ: 7-(N,N-dimethylaminosulfonyl)-4-piperazino-2,1,3-benzoxadiazole.

4. Analytical methods for amino acid metabolites in biological samples

Target Compounds	Biological Sample	Sample Treatment	Derivatization Reagent		Separation Mode		Detection Method		LOD	Recovery	Ref.
Kinurenic acid	Rat plasma	Centrifugation		None	RPLC	FL: 251/398 nm			0.16 nM	97–98%	^[40]
3 Trp metabolites	Mouse plasma and brain	Centrifugation		None	RPLC			UV, FL	0.03–1.33 µM	83–116%	^[41]
6 Trp metabolites	Pig urine, plasma	Centrifugation		None	RPLC			MS	10–100 ng/mL (LOQ)	–	^[42]
Glycated Trp	Chicken plasma	Solvent extraction		None	RPLC			MS	–	–	^[43]
PHP-THβC	Chicken plasma	Cation-exchange resin		None	RPLC			MS	–	–	^[44]
5 Trp and Tyr metabolites	Human urine	Centrifugation		None	RPLC			UV: 220, 280 nm, FL: 280/350, 315/425 nm	–	–	^[45]
DOPAC, HVA	Rat kidney	Microdialysis		Ethylenediamine	Ion exchange chrom atography			FL: 417/495 nm	50, 100 fmol	–	^[46]
Nicotinic acid	Human plasma	Solvent extraction		None	RPLC			MS/MS	6.57 ng/mL (LOQ)	70–72%	^[47]
Glutaric acid, 3-HG	Human urine	Centrifugation		DAABD-AE	RPLC			MS/MS	20–25 nM	94–121%	^[48]
64 amino acid derivatives	Human urine, pancreatic cancer cells	Centrifugation		DmPABr	RPLC			MS/MS	0.11–2192 nM	–	^[49]

PHP-THβC: (1R, 3S)-1-(D-gluco-1, 2, 3, 4, 5-pentahydroxypentyl)-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid, DOPAC: 3,4-dihydroxyphenylacetic acid, HVA: homovanillic acid, 3-HG: 3-hydroxyglutaric acid, DAABD-AE: 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole, DmPABr: dimethylaminophenacyl bromide

5. Analytical methods for perfluorinated carboxylic acids (PFCAs) in biological samples

Target Compounds

Biological Sample

Sample Treatment

Derivatization Reagent

Separation Mode

Detection Method

LOD

Recovery

Ref.

3 PFASs

Human tissues and blood

Solid phase extraction

None

RPLC

3 µg/L

80–101%

^[50]

10 PFASs

Two bivalves shells, soft tissues

Solid phase extraction

None

RPLC

MS/MS

0.05–0.43 ng/g

92–104%

^[51]

18 PFASs

Human urine and serum

Solid phase extraction

None

RPLC

MS/MS

0.1 µg/L

94–104%

^[52]

21 PFASs

Human serum

Solid phase extraction

None

RPLC

MS/MS

0.008–0.19 µg/L

85–114%

^[53]

6 PFASs

Human plasma

µ-SPE

None

RPLC

MS/MS

21–65 ng/L

88–102%

^[54]

6 PFASs

Human serum

Deproteinization

MASH

RPLC

MS/MS

0.07–0.42 µg/L

96–100%

^[55]

11 PFASs

Human blood

Solvent extraction

None

RPLC

MS/MS

0.06–0.14 µg/L

67–112%

^[56]

20 PFASs

Human plasma, BCS

Centrifugation

None

RPLC

MS/MS

0.024–0.096 µg/L (LOQ)

83–103%

^[57]

PFASs: polyfluoroalkyl substances, MASH: 10-methyl-acridone-2-sulfonohydrazide.

6. Analytical methods for α-keto acids and 2-hydroxyglutaric acid (2-HG) in biological samples

Target Compounds

Biological Sample

Sample Treatment

Derivatization Reagent

Separation Mode

Detection Method

LOD

Recovery

Ref.

4 α-Keto acids

Human serum

Centrifugation

OPD

RPLC

FL: 350/410 nm

1 µM

86–109%

^[58]

7 α-Keto acids

Human neutrophil

Centrifugation

OPD

RPLC

FL: 360/415 nm

0.035–0.125 µM

79–108%

^[59]

3 α-Keto acids

Human CML cell

Gel extraction

OPD

RPLC

FL: 360/415 nm

18–40 nM

84–96%

^[60]

6 α-Keto acids

Human CML cell

Centrifugation

DMB

RPLC

FL: 367/446 nm

1.3–5.4 nM

86–118%

^[61]

3 α-Keto acids

Mouse tissue

Acid extraction

OPD

RPLC

5 nM

76–95%

^[62]

10 α-Keto acids

Rat plasma

Centrifugation

O-PFBO

RPLC

MS/MS

0.01–0.25 µM

96–109%

^[63]

3 α-Keto acids

Human plasma

Centrifugation

None

RPLC

MS/MS

0.04 µg/mL

81–98%

^[64]

(R)-2-HG

Human serum

Solid phase extraction

DATAN

RPLC

MS/MS

0.060 µM

31–32%

^[65]

(R)-2-HG

Human urine, cancer tissues

Solvent extraction

TSPC

RPLC

MS/MS

1.2 fmol

88–109%

^[66]

OPD: o-phenylenediamine, DMB: 1,2-diamino-4,5-methylenedioxybenzene, O-PFBO: O-(2,3,4,5,6-pentafluorobenzyl)oxime, DATAN: (+)-o,o’-diacetyl-l-tartaric anhydride, TSPC: N-(p-toluenesulfonyl)-L-phenylalanyl chloride.

7. Analytical methods for 2-aminothiazoline-4-carboxylic acid (ATCA), 2-methylthiazolidine-4-carboxylic acid (MTCA), and 2-thiothiazolidine-4-carboxylic acid (TTCA) in biological samples

Target Compounds

Biological Sample

Sample Treatment

Derivatization Reagent

Separation Mode

Detection Method

LOD

Recovery

Ref.

ATCA

Rat plasma and organ

Solid phase extraction

None

RPLC

MS/MS

–

^[67]

ATCA

Human urine

MISBSE

None

RPLC

MS/MS

5 µg/L

–

^[68]

ATCA

Rat plasma

Solid phase extraction

None

RPLC

MS/MS

12 µg/L

–

^[69]

ATCA

Human postmortem blood

Solid phase extraction

None

HILIC

MS/MS

2.5 µg/L

81–89%

^[70]

ATCA

Human postmortem blood

Solid phase extraction

None

HILIC

MS/MS

9 µg/L (LOQ)

88–96%

^[71]

ATCA

Human post-mortem blood

Liquid-liquid extraction

None

HILIC

MS/MS

0.43 µg/L

86–101%

^[72]

MTCA

Human blood and urine

Centrifugation

Acetic anhydride

RPLC

MS/MS

0.1 mg/L

–

^[73]

TTCA

Urine

Acid extraction

None

RPLC

UV: 271 nm

35 µg/L

78–87%

^[74]

MISBSE: molecularly imprinted stir bar sorption extraction.

8. Analytical methods for other carboxylic acids in biological samples

Target Compounds

Biological Sample

Sample Treatment

Derivatization Reagent

Separation Mode

Detection Method

LOD

Recovery

Ref.

7 Bile acids

Human saliva

SPE and solvent extraction

2-Picolylamine

RPLC

MS/MS

1.5–5.6 fmol

–

^[75]

3 Bile acids, 8 fatty acids

Human plasma and saliva

Solid phase extraction

APBQ

RPLC

MS/MS

0.19–0.51 fmol

–

^[76]

7 Bile acids, 9 fatty acids

Human serum

Solvent extraction

DBCETS

RPLC

FL: 300/395 nm

0.28–0.70 ng/mL

92–102%

^[77]

4 Bile acids

C. bovis

Centrifugation

2-bromo-4′-nitroacetophenone

RPLC

UV: 263 nm

0.25–0.31 ng

94–99%

^[78]

7 Bile acids

Human feces

Solid phase extraction

Phenacyl bromide

RPLC

UV: 254 nm

1.22–1.46 pmol

72–102%

^[79]

Human feces

Solid phase extraction

None

PRLC

MS/MS

–

^[79]

Dihydroxyoxocholestenoic acids

Human CSF and plasma

Solid phase extraction

Isotope-label ed Girard’s P Reagent

RPLC

0.02–0.05 ng/mL

–

^[80]

7 THGC glucuronides

Human urine

Centrifugation

Isotope-labeled DAPPZ

RPLC

MS/MS

0.008–0.16 µg/mL (LOQ)

–

^[81]

Orotic acid

Urine

Dilution

None

RPLC

MS/MS

0.15 µM

–

^[82]

Metabolome

Human urine

Centrifugation

Isotope-label ed DmPABr

RPLC

–

^[83]

Metabolome

Human urine

Centrifugation

Isotope-labeled dansyl hydrazine

RPLC

–

^[84]

APBQ: 1-(3-aminopropyl)-3-bromoquinolinium bromide, DBCETS: 2-(7H-dibenzo[a,g]carbazol-7-yl)ethyl 4-methylbenzenesulfonate, DAPPZ: 1-[(4-dimethylaminophenyl)-carbonyl]piperazine, DmPABr: dimethylaminophenacyl bromide.

This entry is adapted from the peer-reviewed paper 10.3390/molecules25214883

References

Theodoridis: G.A.; Gika, H.G.; Want, E.J.; Wilson, I.D. Liquid chromatography–mass spectrometry based global metabolite profiling: A review. Anal. Chim. Acta 2012, 711, 7–16.
Chen, J.; Wang, W.; Lv, S.; Yin, P.; Zhao, X.; Lu, X.; Zhang, F.; Xu, G. Metabonomics study of liver cancer based on ultra performance liquid chromatography coupled to mass spectrometry with HILIC and RPLC separations. Anal. Chim. Acta 2009, 650, 3–9.
Xu, X.; Roman, J.M.; Issaq, H.J.; Keefer, L.K.; Veenstra, T.D.; Ziegler, R.G. Quantitative Measurement of Endogenous Estrogens and Estrogen Metabolites in Human Serum by Liquid Chromatography-Tandem Mass Spectrometry. Anal. Chem. 2007, 79, 7813–7821.
Tsunoda, M.; Sumida, Y. Liquid Chromatography| Amino Acids. Encyclopedia of Analytical Science (3rd ed.) 2019, 6, 1–11.
Isokawa, M.; Kanamori, T.; Funatsu, T.; Tsunoda, M. Analytical methods involving separation techniques for determination of low-molecular-weight biothiols in human plasma and blood. J. Chromatogr. B. 2014, 964, 103–115.
Tsunoda, M. Recent advances in methods for the analysis of catecholamines and their metabolites. Anal. Bioanal. Chem. 2006, 386, 506–514.
Du, X.-L.; Zhang, H.-S.; Guo, X.-F.; Deng, Y.-H.; Wang, H. 6-Oxy-(acetyl piperazine) fluorescein as a new fluorescent labeling reagent for free fatty acids in serum using high-performance liquid chromatography. J. Chromatogr. A. 2017, 1169, 77–85.
Lu, C.-Y.; Wu, H.-L.; Chen, S.-H.; Kou, H.-S. A Fluorimetric Liquid Chromatography for Highly Sensitive Analysis of Very Long Chain Fatty Acids as Naphthoxyethyl Derivatives. Chromatographia 2000, 51, 315–321.
You, J.; Zhang, W.; Jia, X.; Zhang, Y. An Improved Derivatization Method for Sensitive Determination of Fatty Acids by High-Performance Liquid Chromatography Using 9-(2-hydroxylethyl)-Carbazole as Derivatization Reagent with Fluorescence Detection. Chromatographia 2001, 54, 316–322.
You, J.; Zhang, W.; Zhang, Y. Simple derivatization method for sensitive determination of fatty acids with fluorescence detection by high-performance liquid chromatography using 9-(2-hydroxyethyl)-carbazole as derivatization reagent. Anal. Chim. Acta 2001, 436, 163–172.
Nishikiori, M.; Iizuka, H.; Ichiba, H.; Sadamoto, K.; Fukushima, T. Determination of Free Fatty Acids in Human Serum by HPLC with Fluorescence Detection. J. Chromatogr. Sci. 2015, 53, 537–541.
Onozato, M.; Okanishi, Y.; Akutsu, M.; Okumura, I.; Nemoto, A.; Takano, K.; Sakamoto, T.; Ichiba, H.; Fukushima, T. Alteration in plasma docosahexanoic acid levels following oral administration of ethyl icosapentate to rats. Pract. Lab. Med. 2020, 18, e00143.
Nithipatikom, K.; Pratt, P.F.; Campbell, W.B. Determination of EETs using microbore liquid chromatography with fluorescence detection. Am. J. Physiol. Heart Circ. Physiol. 2000, 279, 857–862.
Bollinger, J.G.; Rohan, G.; Sadilek, M.; Gelb, M.H. LC/ESI-MS/MS detection of FAs by charge reversal derivatization with more than four orders of magnitude improvement in sensitivity. J. Lipid Res. 2013, 54, 3523–3530.
Bollinger, J.G.; Thompson, W.; Lai, Y.; Oslund, R.C.; Hallstrand, T.S.; Sedilek, M.; Turecek, F.; Gelb, M.H. Improved Sensitivity Mass Spectrometric Detection of Eicosanoids by Charge Reversal Derivatization. Anal. Chem. 2010, 82, 6790–6796.
Sun, F.; Choi, A.A.; Wu, R. Systematic Analysis of Fatty Acids in Human Cells with a Multiplexed Isobaric Tag (TMT)-Based Method. J. Proteome Res. 2018, 17, 1606–1614.
Tsukamoto, Y.; Santa, T.; Yoshida, H.; Miyano, H.; Fukushima, T.; Hirayama, K.; Imai, K.; Funatsu, T. Synthesis of the isotope-labeled derivatization reagent for carboxylic acids, 7-(N,N-dimethylaminosulfonyl)-4-(aminoethyl)piperazino-2,1,3-benzoxadiazole (d6) [DBD-PZ-NH2 (D)], and its application to the quantification and the determination of relative amount of fatty acids in rat plasma samples by high-performance liquid chromatography/mass spectrometry. Biomed. Chromatogr. 2006, 20, 358–364.
Tsukamoto, Y.; Santa, T.; Saimaru, H.; Imai, K.; Funatsu, T. Synthesis of benzofurazan derivatization reagents for carboxylic acids and its application to analysis of fatty acids in rat plasma by high-performance liquid chromatography–electrospray ionization mass spectrometry. Biomed. Chromatogr. 2005, 19, 802–808.
Abualhasan, M.N.; Watson, D.G. Tagging Fatty Acids Via Choline Coupling for the Detection of Carboxylic Acid Metabolites in Biological Samples. Curr. Anal. Chem. 2019, 15, 642–647.
Chen, G.-Y.; Zhang, Q. Simultaneous quantification of free fatty acids and acylcarnitines in plasma samples using dansylhydrazine labeling and liquid chromatography–triple quadrupole mass spectrometry. Anal. Bioanal. Chem. 2020, 412, 2841–2849.
Leng, J.; Wang, H.; Zhang, L.; Zhang, J.; Wang, H.; Guo, Y. A highly sensitive isotope-coded derivatization method and its application for the mass spectrometric analysis of analytes containing the carboxyl group. Anal. Chim. Acta 2013, 758, 114–121.
Zhu, Q.-F.; Zhang, Z.; Liu, P.; Zheng, S.-J.; Peng, K.; Deng, Q.-Y.; Zheng, F.; Yuan, B.-F.; Feng, Y.-Q. Analysis of liposoluble carboxylic acids metabolome in human serum by stable isotope labeling coupled with liquid chromatography–mass spectrometry. J. Chromatogr. A. 2016, 1460, 100–109.
Nagy, K.; Jakab, A.; Fekete, J.; Vékey, K. An HPLC-MS Approach for Analysis of Very Long Chain Fatty Acids and Other Apolar Compounds on Octadecyl-Silica Phase Using Partly Miscible Solvents. Anal. Chem. 2004, 76, 1935–1941.
Kotani, A.; Kusu, F.; Takamura, K. New electrochemical detection method in high-performance liquid chromatography for determining free fatty acids. Anal. Chim. Acta 2002, 465, 199–206.
Kotani, A.; Fuse, T.; Kusu, F. Determination of Plasma Free Fatty Acids by High-Performance Liquid Chromatography with Electrochemical Detection. Anal. Biochem. 2000, 284, 65–69.
Morita, H.; Konishi, M. Electrogenerated Chemiluminescence Derivatization Reagents for Carboxylic Acids and Amines in High-Performance Liquid Chromatography Using Tris(2,2′-bipyridine)ruthenium(II). Anal. Chem. 2002, 74, 1584–1589.
Baati, T.; Horcajada, P.; Gref, R.; Couvreur, P.; Serre, C. Quantification of fumaric acid in liver, spleen and urine by high-performance liquid chromatography coupled to photodiode-array detection. J. Pharm. Biomed. Anal. 2011, 56, 758–672.
Chen, H.-C.; Wu, C.; Wu, K.-Y. Determination of the maleic acid in rat urine and serum samples by isotope dilution-liquid chromatography-tandem mass spectrometry with on-line solid phase extraction. Talanta 2015, 136, 9–14.
Lakso, H.; Appelblad, P.; Schneese, J. Quantification of Methylmalonic Acid in Human Plasma with Hydrophilic Interaction Liquid Chromatography Separation and Mass Spectrometric Detection. Clin. Chem. 2008, 54, 2028–2035.
Pellegrini, D.; Onor, M.; Degano, I.; Bramanti, E. Development and validation of a novel derivatization method for the determination of lactate in urine and saliva by liquid chromatography with UV and fluorescence detection. Talanta 2014, 130, 280–287.
Schriewer, A.; Brink, M.; Gianmoena, K.; Cadenas, C.; Hayen, H. Oxalic acid quantification in mouse urine and primary mouse hepatocyte cell culture samples by ion exclusion chromatography-mass spectrometry. J. Chromatogr. B 2017, 1068–1069, 239–244.
Kubota, K.; Fukushima, T.; Yuji, R.; Miyano, H.; Hirayama, K.; Santa, T.; Imai, K. Development of an HPLC-fluorescence determination method for carboxylic acids related to the tricarboxylic acid cycle as a metabolome tool. Biomed. Chromatogr. 2005, 19, 788–795.
Niu, H.; Chen, Y.; Xie, J.; Chen, X.; Bai, J.; Wu, J.; Liu, D.; Ying, H. Ion-Exclusion Chromatography Determination of Organic Acid in Uridine 5′-Monophosphate Fermentation Broth. J. Chromatogr. Sci. 2012, 50, 709–713.
Halko, R.; Hukelová, I. Single-Run Separation and Determination of Aliphatic and Aromatic Carboxylic Acids in Wine and Human Urine Samples by Ion-Exclusion Chromatography. Chromatographia 2014, 77, 1037–1046.
Todoroki, K.; Hashimoto, H.; Machida, K.; Itoyama, M.; Hayama, T.; Yoshida, H.; Nohta, H.; Nakashima, M.; Yamaguchi, M. Fully automated reagent peak-free liquid chromatography fluorescence analysis of highly polar carboxylic acids using a column-switching system and fluorous scavenging derivatization. J. Sep. Sci. 2013, 36, 232–238.
Michopoulos, F.; Whalley, N.; Theodoridis, G.; Wilson, I.D.; Dunkley, T.P.J.; Critchlow, S.E. Targeted profiling of polar intracellular metabolites using ion-pair-high performance liquid chromatography and -ultra high performance liquid chromatography coupled to tandem mass spectrometry: Applications to serum, urine and tissue extracts. J. Chromatogr. A. 2014, 1349, 60–68.
Guo, L.; Worth, A.J.; Mesaros, C.; Snyder, N.W.; Glickson, J.D.; Blair, I.A. Diisopropylethylamine/hexafluoroisopropanol-mediated ion-pairing UHPLC-MS for phosphate and carboxylate metabolite analysis: Utility for studying cellular metabolism. Rapid Commun. Mass Spectrom. 2016, 30, 1835–1845.
Buescher, J.M.; Moco, S.; Sauer, U.; Zamboni, N. Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Fast and Robust Quantification of Anionic and Aromatic Metabolites. Anal. Chem. 2010, 82, 4403–4412.
Nemkov, T.; Sun, K.; Reisz, J.A.; Yoshida, T.; Dunham, A.; Wen, E.Y.; Wen, A.Q.; Roach, R.C.; Hansen, K.C.; Xia, Y.; et al. Metabolism of citrate and Other carboxylic acids in erythrocytes as a Function of Oxygen saturation and refrigerated storage. Front. Med. 2017, 4, 175.
Fukushima, T.; Sone, Y.; Mitsuhashi, S.; Tomita, M.; Toyo’oka, T. Alteration of Kynurenic Acid Concentration in Rat Plasma Following Optically Pure Kynurenine Administration: A Comparative Study Between Enantiomers. Chirality 2009, 21, 468–472.
Cseh, E.K.; Veres, G.; Szentirmai, M.; Nánási, N.; Szatmári, I.; Fülöp, F.; Vécsei, L.; Zádori, D. HPLC method for the assessment of tryptophan metabolism utilizing separate internal standard for each detector. Anal. Biochem. 2019, 574, 7–14.
Brunius, C.; Vidanarachchi, J.K.; Tomankova, J.; Lundström, K.; Andersson, K.; Zamaratskaia, G. Skatole metabolites in urine as a biological marker of pigs with enhanced hepatic metabolism. Animal 2016, 10, 1734–1740.
Kita, K.; Kawashima, Y.; Makino, R.; Namauo, T.; Ogawa, S.; Muraoka, H.; Fujimura, S. Detection of Two Types of Glycated Tryptophan Compounds in the Plasma of Chickens Fed Tryptophan Excess Diets. J. Poult. Sci. 2013, 50, 138–142.
Makino, R.; Kita, K. Half-life of Glycated Tryptophan in the Plasma of Chickens. J. Poult. Sci. 2018, 55, 117–119.
Valko-Rokytovská, M.; Hubková, B.; Birková, A.; Mašlanková, J.; Stupák, M.; Zábavníková, M.; Cižmárová, B.; Mareková, M. Specific Urinary Metabolites in Malignant Melanoma. Medicina 2019, 55, 145.
Tsunoda, M.; Mitsuhashi, K.; Masuda, M.; Imai, K. Simultaneous determination of 3,4-dihydroxyphenylacetic acid and homovanillic acid using high performance liquid chromatography-fluorescence detection and application to rat kidney microdialysate. Anal. Biochem. 2002, 307, 153–158.
Huang, W.-H.; Hu, K.; Shao, L.; Chen, Y.; Zhang, W.; Zhou, H.-H.; Tan, Z.-R. Development and validation of a method for the determination of nicotinic acid in human plasma using liquid chromatography-negative electrospray ionization tandem mass spectrometry and its application to a bioequivalence study. Anal. Methods 2014, 6, 8258–8267.
Al-Dirbashi, O.Y.; Santa, T.; Al-Qahtani ,K.; Al-Amoudi, M.; Rashed, M.S. Analysis of organic acid markers relevant to inherited metabolic diseases by ultra-performance liquid chromatography/tandem mass spectrometry as benzofurazan derivatives. Rapid Commun. Mass Spectrom. 2007, 21, 1984–1990.
Willacey, C.C.W.; Naaktgeboren, M.; Moreno, E.L.; Wegrzyn, A.B.; Es, D.; Karu, N.; Fleming, R.M.T.; Harms, A.C.; Hankemeier, T. LC-MS/MS analysis of the central energy and carbon metabolites in biological samples following derivatization by dimethylaminophenacyl bromide. J. Chromatogr. A. 2019, 1608, 460413.
Maestri, L.; Negri, S.; Ferrari, M.; Ghittori, S.; Fabris, F.; Danesino, P.; Imbriani, M. Determination of perfluorooctanoic acid and perfluorooctanesulfonate in human tisseues by liquid chromatography/single quadrupole mass spectrometry. Rapid Commun. Mass Spectrom. 2006, 20, 2728–2734.
Wang, L.; Sun, H.; Yang, L.; He, C.; Wu, W.; Sun, S. Liquid chromatography/mass spectrometry analysis of perfluoroalkyl carboxylic acids and perfluorooctanesulfonate in bivalve shells: Extraction method optimization. J. Chromatogr. A. 2010, 1217, 436–442.
Kato, K.; Kalathil, A.A.; Patel, A.M.; Ye, X.; Calafat, A.M. Per- and polyfluoroalkyl substances and fluorinated alternatives in urine and serum by on-line solid phase extraction-liquid chromatogtaphy-tandem mass spectrometry. Chemosphere 2018, 209, 338–345.
Gao, K.; Gao, Y.; Li, Y.; Fu, J.; Zhang, A. A rapid and fully automatic method for the accurate determination of a wide carbon-chain range of per- and polyfluoroalkyl substances (C4–C18) in human serum. J. Chromatogr. A. 2016, 1471, 1–10.
Lashgari, M.; Lee, H.K. Micro-solid phase extraction of perfluorinated carboxylic acids from human plasma. J. Chromatogr. A. 2016, 1432, 7–16.
Zhang, S.; Ji, Z.; Sun, Z.; Li, M.; Sheng, C.; Yue, M.; Yu, Y.; Chen, G.; You, J. Stable isotope labeling assisted liquid chromatography-tandem mass spectrometry for the analysis of perfluorinated carboxylic acids in serum samples. Talanta 2017, 166, 255–261.
Liu, L.; She, J.; Zhang, X.; Zhang, J.; Tian, M.; Huang, Q.; Eqani, S.A.M.A.S.; Shen, H. Online background cleanup followed by high-performance liquid chromatography with tandem mass spectrometry for the analysis of perfluorinated compounds in human blood. J. Sep. Sci. 2015, 38, 247–253.
Harrington, L.M. Analysis of perfluoroalkyl and polyfluoroalkyl substances in serum and plasma by solvent precipitation-isotope dilution-direct injection-LC/MS/MS. Anal. Methods, 2017, 9, 473–481.
Pailla, K.; Blonde-Cynober, F.; Aussel, C.; Bandt, J.; Cynober, L. Branched-Chain Keto-Acids and Pyruvate in Blood: Measurement by HPLC with Fluorimetric Detection and Changes in Older Subjects. Clin. Chem. 2000, 46, 848–853.
Mühling, J.; Fuchs, M.; Campos, M.E.; Gonter, J.; Engel, J.M.; Sablotzki, A.; Menges, T.; Weiss, S.; Dehne, M.G.; Krüll, M.; et al. Quantitative determination of free intracellular α-keto acids in neutrophils. J. Chromatogr. B. 2003, 789, 383–392.
Hattori, A.; Ito, T.; Tsunoda, M. Analysis of Branched-Chain Keto Acids in Cell Extracts by HPLC-Fluorescence Detection. Chromatography 2017, 38, 129–133.
Fujiwara, T.; Hattori, A.; Ito, T.; Funatsu, T.; Tsunoda, M. Analysis of intracellular α-keto acids by HPLC with fluorescence detection. Anal. Methods 2020, 12, 2555–2559.
Olson, K.C.; Chen, G.; Lynch, C.J. Quantification of branched-chain keto acids in tissue by ultra fast liquid chromatography-mass spectrometry. Anal. Biochem. 2013, 439, 116–122.
Noguchi, K.; Mizukoshi, T.; Miyano, H.; Yamada, N. Development of a New LC-MS/MS Method for the Quantification of Keto Acids. Chromatography 2014, 35, 117–123.
Li, R.; Liu, P.; Liu, P.; Tian, Y.; Hua, Y.; Gao, Y.; He, H.; Chen, J.; Zhang, Z.; Huang, Y. A novel liquid chromatography tandem mass spectrometry method for simultaneous determination of branched-chain amino acids and branched-chain α-keto acids in human plasma. Amino Acids 2016, 48, 1523–1532.
Poinsignon, V.; Mercier, L.; Nakabayashi, K.; David, M.D.; Lalli, A.; Penard-Lacronique, V.; Quivoron, C.; Saada, V.; Botton, S.D.; Broutin, S.; et al. Quantitation of isocitrate dehydrogenase (IDH)-induces D and L enantiomers of 2-hydroxyglutaric acid in biological fluids by a fully validated liquid tandem mass spectrometry method, suitable for clinical applications. J. Chromatogr. B. 2016, 1022, 290–297.
Cheng, Q.-Y.; Xiong, J.; Huang, W.; Ma, Q.; Ci, W.; Feng, Y.-Q.; Yuan, B.-F. Sensitive Determination of Onco-metabolites of D- and L-2-hydroxyglutarate Enantiomers by Chiral Derivatization Combined with Liquid Chromatography/Mass Spectrometry Analysis. Sci. Rep. 2015, 5, 15217.
Petrikovics, I.; Thompson, D.E.; Rockwood, G.A.; Logue, B.A.; Martin, S.; Jayanna, P.; Yu, J.C.C. Organ-distribution of the metabolite 2-aminotiazoline-4-carboxylic acid in a rat model following cyanide exposure. Biomarkers 2011, 16, 686–690.
Jackson, R.; Petrikovics, I.; Lai, E.P.C.; Yu, J.C.C. Molecularly imprinted polymer stir bar sorption extraction and electrospray ionization tandem mass spectrometry for determination of 2-aminothiazoline-4-carboxylic acid as a marker for cyanide exposure in forensic urine analysis. Anal. Methods 2010, 2, 552–557.
Petrikovics, I.; Yu, J.C.C.; Thompson, D.E.; Jayanna, P.; Logue, B.A.; Nasr, J.; Bhandari, R.K.; Baskin, S.I.; Rockwood, G. Plasma persistence of 2-aminothiazoline-4-carboxylic acid in rat system determined by liquid chromatography tandem mass spectrometry. J. Chromatogr. B. 2012, 891–892, 81–84.
Luliński, P.; Giebułtowicz, J.; Wroczyński, P.; Maciejewska, D. A highly selective molecularly imprinted sorbent for extraction of 2-aminothiazoline-4-carboxylic acid – Synthesis, characterization and application in post-mortem whole blood analysis. J. Chromatogr. A. 2015, 1420, 16–25.
Giebułtowicz, J.; Sobiech, M.; Rużycka, M.; Luliński, P. Theoretical and experimental approach to hydrophilic interaction dispersive solid-phase extraction of 2-aminothiazoline-4-carboxylic acid from human post-mortem blood. J. Chromatogr. A. 2019, 1587, 61–72.
Giebułtowicz, J.; Rużycka, M.; Fudalej, M.; Krajewski, P.; Wroczyński, P. LC-MS/MS method development and validation for quantitative analysis of 2-aminothiazoline-4-carboxylic acid – a new cyanide exposure marker in post mortem blood. Talanta 2016, 150, 586–592.
Reischl, R.J.; Bicker, W.; Keller, T.; Lamprecht, G.; Lindner, W. Occurrence of 2-methyltiazoline-4-carboxylic acid, a condensation product of cysteine and acetaldehyde, in human blood as a consequence of ethanol consumption. Anal. Bioanal. Chem. 2012, 404, 1779–1787.
Chen, C.-W.; Shih, T.-S.; Li, C.-C.; Chou, J.-S. High Performance Liquid Chromatographic Determination of 2-Thiotiazolidine-4-Carboxylic Acid as a Marker of Occupational Exposure to Carbon Disulfide. Chromatographia 2001, 53, 665–668.
Higashi, T.; Ichikawa, T.; Inagaki, S.; Min, J.Z.; Fukushima, T.; Toyo’oka, T. Simple and practical derivatization procedure for enhanced detection of carboxylic acids in liquid chromatography-electrospray ionization-tandem mass spectrometry. J. Pharm. Biomed. Anal. 2010, 52, 809–818.
Mochizuki, Y.; Inagaki, S.; Suzuki, M.; Min, J.Z.; Inoue, K.; Todoroki, K.; Toyo’oka, T. A novel derivatization reagent possessing a bromoquinolinium structure for biological carboxylic acids in HPLC-ESI-MS/MS. J. Sep. Sci. 2013, 36, 1883–1889.
Li, G.-L.; Chen, G.; Liu, Y.-Q.; Jing, N.-H.; You, J.-M. A sensitive and selective HPLC-FLD method with fluorescent labeling for simultaneous detection of bile acid and free fatty acid in human serum. J. Chromatogr. B. 2012, 895–896, 191–195.
Shi, Y.; Xiong, J.; Sun, D.; Liu, W.; Wei, F.; Ma, S.; Lin, R. Simultaneous quantification of the major bile acids in Artificial Calculus bovis by high-performance liquid chromatography with precolumn derivatization and its application in quality control. J. Sep. Sci. 2015, 38, 2753–2762.
Kakiyama, G.; Muto, A.; Takei, H.; Nittono, H.; Murai, T.; Kurosawa, T.; Hofmann, A.F.; Pandak, W.M.; Bajaj, J.S. A simple and accurate HPLC method for fecal bile acid profile in healthy and cirrhotic subjects: Validation by GC-MS and LC-MS. J. Lipid. Res. 2014, 55, 978–990.
Abdel-Khalik, J.; Crick, P.J.; Yutuc, E.; DeBarber, A.E.; Duell, P.B.; Steiner, R.D.; Laina, I.; Wang, Y.; Griffiths, W.J. Identification of 7α,24-dihydroxy-3-oxocholest-4-en-26-oic and 7α,25-dihydroxy-3-oxocholest-4-en-26-oic acids in human cerebrospinal fluid and plasma. Biochimie 2018, 153, 86–98.
Matsumoto, T.; Yamazaki, W.; Jo, A.; Ogawa, S.; Mitamura, K.; Ikegawa, S.; Higashi, T. A Method for Quantification of Tetrahydroglucocorticoid Glucuronides in Human Urine by LC/MS/MS with Isotope-coded Derivatization. Anal. Sci. 2018, 34, 1003–1009.
La Marca, G.; Casetta, B.; Zammarchi, E. Rapid determination of orotic acid in urine by a fast liquid chromatography/tandem mass spectrometric method. Rapid Commun. Mass Spectrom. 2003, 17, 788–793.
Guo, K.; Li, L. High-Performance Isotope Labeling for Profiling Carboxylic Acid-Containing Metabolites in Biofluids by Mass Spectrometry. Anal. Chem. 2010, 82, 8789–8793.
Zhao, S.; Li, L. Dansylhydrazine Isotope Labeling LC-MS for Comprehensive Carboxylic Acid Submetabolome Profiling. Anal. Chem. 2018, 90, 13514–13522.

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