Validation of a high performance liquid chromatography-tandem mass spectrometry method for β-hydroxy fatty acids as environmental markers of lipopolysaccharide

Chiominto, Alessandra; Marcelloni, Anna Maria; Tranfo, Giovanna; Paba, Emilia; Paci, Enrico

doi:10.1016/j.chroma.2014.01.052

Cited by 4 publications

(2 citation statements)

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“…The samples were analyzed by HPLC-MS/MS technique for detection of 3-OH C 12 and 3-OH C 14 according to the procedure previously published [12]. Briefly, fatty acids were extracted, after alcaline hydrolysis, with n-hexane and extracts were dried in evaporator for 30 minutes at 60°C.…”

Section: Hplc-ms/ms Methodsmentioning

confidence: 99%

“…Besides, the use of the tandem mass spectrometry detection increases its sensitivity and specificity. In a previous paper we presented a new analytical method by HPLC-MS/MS to quantify β-hydroxy-dodecanoic (3-OH C 12 , lauric acid) and β-hydroxy-tetradecanoic (3-OH C 14 , myristic acid) acids and a pilot study was carried out on four ATCC Gram-negative bacteria and two fungi [12]. The choice of these acids is due to the fact that previous reports indicate that shorter-chains (C 10 -C 14 ) β-hydroxyfatty acids are positively correlated with endotoxin activity in the Limulus bioassay while longer-chain (C 16 -C 18 ) 3-OHFAs tend to have lower or even no correlation [13].…”

Section: Introductionmentioning

confidence: 99%

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Endotoxin Analysis: Correlation Between Biological and Chemical Methods

Kong¹,

Chiominto²,

Marcelloni³

et al. 2019

BJSTR

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Assessment of occupational exposure to airborne endotoxins was studied for several years but still different procedures are used for sampling and analysis. Among analytical methods LAL test is the most used. A different approach is represented by chemical analyses, based on the detection of the β-hydroxy fatty acids, as markers of lipopolysaccharide (LPS). In this study an analytical method using HPLC-MS/MS was applied to quantify the content of β-hydroxy-dodecanoic (3-OH C 12 ) and β-hydroxytetradecanoic (3-OH C 14 ) acids in thirteen Gram-negative bacterial strains. In order to compare biological and chemical results cell suspensions were analyzed also by Kinetic Chromogenic LAL test. The results obtained by HPLC-MS/MS show that three samples did not contain 3-OH C 12 at all, while 3-OH C 14 was measured in a concentration range from 11.1 to 4007.5µg/L. Endotoxin concentration measured by LAL assay was in the range of 10 2 -10 3 µg/L. Pearson's test showed that the sum of fatty acids by HPLC-MS/MS correlates positively with the LAL Test (r= 0.79) confirming that it can be considered a reliable marker of endotoxin contamination. Principal Component Analysis (PCA) confirms above results and provides additional information grouping microorganisms related with each other on the basis of their biochemical features, habitat and disease spectra.

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Section: Hplc-ms/ms Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%