Highly Sensitive Fluorescence Detection of Daptomycin in Murine Samples through Derivatization with 2,3-Naphthalenedialdehyde

Yamamoto, Takuhei; Sakamoto, Kousuke; Esaka, Yukihiro; Uno, Bunji

doi:10.2116/analsci.20n010

Cited by 4 publications

(2 citation statements)

References 19 publications

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“…The asparagine and glutamate concentrations were also measured by the modified High Performance Liquid Chromatography (HPLC) fluorescence detection method 10 . After the enzyme reaction, samples were centrifuged at 4°C and 20 000 g for 60 minutes with a 3 K molecular weight cut‐off filter.…”

Section: Methodsmentioning

confidence: 99%

In vitro functional analysis of four variants of human asparagine synthetase

Matsumoto

Kawashima

Yamamoto

et al. 2021

J of Inher Metab Disea

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The loss‐of‐function variants of the human asparagine synthetase (ASNS) gene cause asparagine synthetase deficiency (ASNSD). Diagnosis of ASNSD requires genetic tests because a specific biochemical diagnostic for ASNSD is not available. There are a few reports describing the functional evaluation of ASNS variants. Therefore, in vitro methods are needed to evaluate the detected variants in patients. In this report, five types of human ASNS proteins (wild‐type and our reported four variants: p.Leu145Ser, p.Leu247Trp, p.Val489Asp, and p.Trp541Cysfs*5) were expressed in silkworm using a baculoviral expression system. An enzymatic activity assay of ASNS was performed, and the concentration of asparagine by ninhydrin and High Performance Liquid Chromatography methods using the purified recombinant proteins was measured. We established ASNS deficient HEK293 cells using the CRISPR/Cas9 method and evaluated the growth of cells without asparagine after transduction of ASNS variants with a lentiviral expression system. The four ASNS variants displayed significantly low enzymatic activity. The ASNS deficient HEK293 cells transduced with wild‐type ASNS grew without asparagine, whereas cells transduced with the variants did not grow or showed significantly slower growth than cells transduced with wild‐type ASNS. Herein, we established a method for evaluating the enzymatic activity of the recombinant human ASNS variants. The results of the cell‐based assay corroborated the results of the enzymatic activity. These methods should enable the evaluation of the pathogenicity of ASNS variants.

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Section: Methodsmentioning

confidence: 99%

In vitro functional analysis of four variants of human asparagine synthetase

Matsumoto

Kawashima

Yamamoto

et al. 2021

J of Inher Metab Disea

Self Cite

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“…High performance liquid chromatography (HPLC) is a general separation method used in various research fields and is applied to the separations of diverse analytes with different separation modes, such as reversed-phase, normal phase, hydrophobic interactions, and ion-exchange. The analysis of various compounds using HPLC has been reported even today [1][2][3][4][5][6]. Conventional HPLC separation is generally performed at an ambient temperature.…”

Section: Introductionmentioning

confidence: 99%

Effect of Organic Modifier on the Retention of Low-Molecular-Weight Organic Compounds in Low-Temperature HPLC Using a Liquid CO<sub>2</sub> Mobile Phase and an Octadecyl Stationary Phase

et al. 2021

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The retention behaviors of various low-molecular-weight compounds in low-temperature HPLC with an ODS (octadecyl silica) column and liquid CO 2 mobile phase containing methanol or acetonitrile as modifiers were investigated. The addition of methanol/acetonitrile reduced the retention of xanthone, which was dominated by the liquid CO 2 content, independent of the modifier. Additionally, the retention of polycyclic aromatic hydrocarbons (PAHs) was reduced by the addition of methanol/acetonitrile. Because acetonitrile resulted in a rapid reduction in the retention compared with methanol, it was suggested that the CH- interactions have an important role in the retention of PAHs in the C18 stationary phase. Notably, the retention of alkylbenzenes were only slightly affected by the addition of methanol/acetonitrile. Acetonitrile slightly increased the retention of alkylbenzenes with longer alkyl chains than those with shorter alkyl chains. It is suggested that the interaction of the phenyl moiety in alkylbenzenes with C18 chains was reduced by the increase in the acetonitrile content, and enhanced in the case of the alkyl chain moieties. The retentions of alkyl benzenes were approximately compensated, when the alkyl chain length was relatively short.

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Derivatization in liquid chromatography

Poole

2023

Liquid Chromatography

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Highly Sensitive Fluorescence Detection of Daptomycin in Murine Samples through Derivatization with 2,3-Naphthalenedialdehyde

Cited by 4 publications

References 19 publications

In vitro functional analysis of four variants of human asparagine synthetase

In vitro functional analysis of four variants of human asparagine synthetase

Effect of Organic Modifier on the Retention of Low-Molecular-Weight Organic Compounds in Low-Temperature HPLC Using a Liquid CO<sub>2</sub> Mobile Phase and an Octadecyl Stationary Phase

Derivatization in liquid chromatography

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