Measurement of free and total sialic acid by isotopic dilution liquid chromatography tandem mass spectrometry method

The aim of this work was to measure N-acetylneuraminic acid (Neu5Ac) in milk-based infant formulas. The analysis was performed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The total Neu5Ac were released using trichloroacetic acid and hydrochloric acid and purified using a HLB column. The linearity from 0.05 to 5.0 lg/mg Neu5Ac was adequate. Sialic acid recoveries ranged from 91.8% to 112.4%. The detection and quantification limits (limit of detection, 0.01 lg Neu5Ac/mg; limit of quantitation, 1.08 lg Neu5Ac/mg) were low enough to determine the sialic acid in infant formulas. The validated method is highly reproducible and sensitive, and it is easy to perform.

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“…; Tebani et al . ) was consistent with our result. We believe sialic acid can be analysed very well in ESI + .…”

Section: Resultssupporting

confidence: 94%

Determining Neu5Ac in infant formula with ultra‐performance liquid chromatography–tandem mass spectrometry

Chun

Xie

et al. 2015

Int J of Dairy Tech

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“…For Neu5Ac, although reverse-phase chromatography was reported in some methods, the poor retention could cause deteriorated peak shape, high matrix effect, and low sensitivity. A derivatization method was recently reported to increase hydrophobicity and achieve sufficient retention on a reverse-phase column [22], but this additional step lengthens the sample preparation time and increases assay complexity.…”

Section: Resultsmentioning

confidence: 99%

“…These methods are limited by specificity and/or sensitivity, and they cannot be applied for definitive quantitation. Recently, additional methods for Neu5Ac were described based on liquid chromatography–tandem mass spectrometry (LC-MS/MS) [21,22]. These methods achieve the desired specificity.…”

Section: Introductionmentioning

confidence: 99%

Quantitative hydrophilic interaction chromatography–mass spectrometry analysis of N-acetylneuraminic acid and N-acetylmannosamine in human plasma

Shi

Meng

et al. 2015

Journal of Chromatography B

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N-Acetylneuraminic acid (Neu5Ac or NANA) is the most predominant sialic acid in mammals. As a terminal component in many glycoproteins and glycolipids, sialic acid is believed to be an important biomarker related to various diseases. Its precursor, N-acetylmannosamine (ManNAc), is being investigated as a potential treatment for GNE myopathy. In this work, we developed two highly sensitive and selective liquid chromatography–tandem mass spectrometry (LC-MS/MS) methods for the quantitation of ManNAc and free Neu5Ac in human plasma. A fit-for-purpose approach was adopted during method validation and sample analysis. To measure the endogenous compounds and overcome the interference from plasma samples, a surrogate matrix that contained 5% bovine serum albumin (BSA) was used for the preparation of calibration standards and certain levels of quality control (QC) samples. QC samples at higher concentrations were prepared in the authentic matrix (human plasma) to best mimic incurred samples. For both methods, an Ostro 96-well phospholipid removal plate was used for sample extraction, which efficiently removed the phospholipids from the plasma samples prior to LC injection, eliminated matrix effect, and improved sensitivity. Chromatographic separation was achieved using hydrophilic interaction chromatography (HILIC) and gradient elution in order to retain the two polar compounds. The lower limit of quantitation (LLOQ) for ManNAc and Neu5Ac was 10.0 and 25.0 ng/mL, respectively. The overall accuracy of the two assays was no less than 91.7% based on three levels of QC samples. Inter- and intra-run precision (coefficient of variation [%CV]) across three analytical runs was less than 6.7% for ManNAc and less than 10.8% for Neu5Ac. These methods have been validated to support clinical studies.

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“…Quantification of total sialic acids or glycolipid-bound sialic acids in serum is helpful to improve the accuracy of clinical diagnoses and therapies. Detection methods have been established for quantification of sialic acids, including colorimetric assay [99], fluorometric assay [100,101], high performance liquid chromatography (HPLC) [102,103], fluorescence [104][105][106], and mass spectrometry (MS) [107][108][109]. Among those methods, HPLC and MS are widely used [110,111].…”

Section: Sialic Acids Quantificationmentioning

confidence: 99%

Biological Functions and Analytical Strategies of Sialic Acids in Tumor

Zhou

Yang

Guan

2020

Cells

133

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Sialic acids, a subset of nine carbon acidic sugars, often exist as the terminal sugars of glycans on either glycoproteins or glycolipids on the cell surface. Sialic acids play important roles in many physiological and pathological processes via carbohydrate-protein interactions, including cell–cell communication, bacterial and viral infections. In particular, hypersialylation in tumors, as well as their roles in tumor growth and metastasis, have been widely described. Recent studies have indicated that the aberrant sialylation is a vital way for tumor cells to escape immune surveillance and keep malignance. In this article, we outline the present state of knowledge on the metabolic pathway of human sialic acids, the function of hypersialylation in tumors, as well as the recent labeling and analytical techniques for sialic acids. It is expected to offer a brief introduction of sialic acid metabolism and provide advanced analytical strategies in sialic acid studies.

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Measurement of free and total sialic acid by isotopic dilution liquid chromatography tandem mass spectrometry method

Cited by 31 publications

References 25 publications

Determining Neu5Ac in infant formula with ultra‐performance liquid chromatography–tandem mass spectrometry

Determining Neu5Ac in infant formula with ultra‐performance liquid chromatography–tandem mass spectrometry

Quantitative hydrophilic interaction chromatography–mass spectrometry analysis of N-acetylneuraminic acid and N-acetylmannosamine in human plasma

Biological Functions and Analytical Strategies of Sialic Acids in Tumor

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