High-performance liquid chromatography determination of ketone bodies in human plasma by precolumn derivatization with p-nitrobenzene diazonium fluoroborate

Yamato, Susumu; Shinohara, Kumiko; Nakagawa, Saori; Kubota, Ai; Inamura, Katsushi; Watanabe, Gen; Hirayama, Satoshi; Miida, Takashi; Ohta, Shin

doi:10.1016/j.ab.2008.09.017

Cited by 20 publications

(9 citation statements)

References 16 publications

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“…The abundance of each lipid class was quantified using a standard charring and copper-based densitometric assay (Bitman and Wood, 1982; Ruiz and Ochoa, 1997). Ketone bodies were quantified using a Hitachi HPLC equipped with a Capcell Pak 5 μm C8 UG 120 Å, LC Column (150×4.6 mm) as described in detail (Yamato et al, 2009). Briefly, liver lysates were mixed and incubated with the diazo reagent and introduced to HPLC to quantify the acetoacetate.…”

Section: Methodsmentioning

confidence: 99%

Polyunsaturated fatty acyl-coenzyme As are inhibitors of cholesterol biosynthesis in zebrafish and mice

Karanth

Tran

Kuberan

et al. 2013

Disease Models &Amp; Mechanisms

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SUMMARYLipid disorders pose therapeutic challenges. Previously we discovered that mutation of the hepatocyte β-hydroxybutyrate transporter Slc16a6a in zebrafish causes hepatic steatosis during fasting, marked by increased hepatic triacylglycerol, but not cholesterol. This selective diversion of trapped ketogenic carbon atoms is surprising because acetate and acetoacetate can exit mitochondria and can be incorporated into both fatty acids and cholesterol in normal hepatocytes. To elucidate the mechanism of this selective diversion of carbon atoms to fatty acids, we fed wild-type and slc16a6a mutant animals high-protein ketogenic diets. We find that slc16a6a mutants have decreased activity of the rate-limiting enzyme of cholesterol biosynthesis, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (Hmgcr), despite increased Hmgcr protein abundance and relative incorporation of mevalonate into cholesterol. These observations suggest the presence of an endogenous Hmgcr inhibitor. We took a candidate approach to identify such inhibitors. First, we found that mutant livers accumulate multiple polyunsaturated fatty acids (PUFAs) and PUFA-CoAs, and we showed that human HMGCR is inhibited by PUFA-CoAs in vitro. Second, we injected mice with an ethyl ester of the PUFA eicosapentaenoic acid and observed an acute decrease in hepatic Hmgcr activity, without alteration in Hmgcr protein abundance. These results elucidate a mechanism for PUFA-mediated cholesterol lowering through direct inhibition of Hmgcr.

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Section: Methodsmentioning

confidence: 99%

Polyunsaturated fatty acyl-coenzyme As are inhibitors of cholesterol biosynthesis in zebrafish and mice

Karanth

Tran

Kuberan

et al. 2013

Disease Models &Amp; Mechanisms

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“…Several methods are available to measure b-HB concentrations in blood (5)(6)(7)(8). Most are enzymatic methods measuring the concentration of the enantiomer D-b-HB in plasma.…”

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confidence: 99%

Accuracy of the Precision^® point-of-care ketone test examined by liquid chromatography tandem-mass spectrometry (LC-MS/MS) in the same fingerstick sample

Janssen

Hendrickx

Poel

et al. 2010

Clinical Chemistry and Laboratory Medicine

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“…Examples of existing crystalline aryl diazonium are p-nitro benzenediazonium tetrafluoroborate [25] and formyl benzenediazonium hexafluorophsophate [26]. CDNBD applications in both analytical and syntheses utilize reagent solution in situ.…”

Section: Introductionmentioning

confidence: 99%

Modeling of 4-Carboxyl-2,6-dinitrobenzenediazonium Ion (CDNBD) Hydrolysis Through Addition of Water and Alkaline Buffer in a Strongly Acidic Medium

Thomas

Bioku

2015

J Solution Chem

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The transition from aryl diazonium reagent solution to a crystalline form has substantial merits, namely augmented stability and versatility. Absorbance decay data was from photometric titration with hydroxyl ion, supplied incrementally by water and alkaline buffer (pH 12.0), to 4-carboxyl-2,6-dinitrobenzenediazonium ion (CDNBD) and simpler analogs in a strongly acidic medium. The data were fitted to nested models (mono-and bi-exponential decay). Akaike's Information Criterion was used for statistical model comparison. Preferred model identification shows simpler diazonium analogs are less reactive towards hydroxyl ion when alkaline buffer solution is the diluting medium. In contrast, CDNBD hydrolysis is faster when an alkaline diluting medium is added, owing to profound positive effect of strong electron withdrawing groups on its electrophilic reactivity. Acidic diluting medium was shown, unambiguously, as a critical requirement for maintaining CDNBD, in an acidic solution as the cation species. Key input process variables were reliably predicted, using model parameters like V 1=2 and V T 1=2 , thus giving direction for optimal synthesis of crystalline CDNBD.

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High-performance liquid chromatography determination of ketone bodies in human plasma by precolumn derivatization with p-nitrobenzene diazonium fluoroborate

Cited by 20 publications

References 16 publications

Polyunsaturated fatty acyl-coenzyme As are inhibitors of cholesterol biosynthesis in zebrafish and mice

Polyunsaturated fatty acyl-coenzyme As are inhibitors of cholesterol biosynthesis in zebrafish and mice

Accuracy of the Precision^® point-of-care ketone test examined by liquid chromatography tandem-mass spectrometry (LC-MS/MS) in the same fingerstick sample

Modeling of 4-Carboxyl-2,6-dinitrobenzenediazonium Ion (CDNBD) Hydrolysis Through Addition of Water and Alkaline Buffer in a Strongly Acidic Medium

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High-performance liquid chromatography determination of ketone bodies in human plasma by precolumn derivatization with p-nitrobenzene diazonium fluoroborate

Cited by 20 publications

References 16 publications

Polyunsaturated fatty acyl-coenzyme As are inhibitors of cholesterol biosynthesis in zebrafish and mice

Polyunsaturated fatty acyl-coenzyme As are inhibitors of cholesterol biosynthesis in zebrafish and mice

Accuracy of the Precision® point-of-care ketone test examined by liquid chromatography tandem-mass spectrometry (LC-MS/MS) in the same fingerstick sample

Modeling of 4-Carboxyl-2,6-dinitrobenzenediazonium Ion (CDNBD) Hydrolysis Through Addition of Water and Alkaline Buffer in a Strongly Acidic Medium

Contact Info

Product

Resources

About

Accuracy of the Precision^® point-of-care ketone test examined by liquid chromatography tandem-mass spectrometry (LC-MS/MS) in the same fingerstick sample