Determination of an unstable pentapeptide, monocyte locomotion inhibitory factor, in dog blood by LC–MS/MS: Application to a pharmacokinetic study

Jiang, Huafang; Li, Ji; Zhao, Xin; Zhou, Yu; Li, You; Sun, Fanlu; Hu, Xun; Huang, Junkai; Rui, Yao-Cheng; Fan, Guorong

doi:10.1016/j.jpba.2013.05.013

Cited by 10 publications

(2 citation statements)

References 36 publications

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“…This value is in the range of LLOQ (2–300 ng/mL) published in other studies for the determination of various pharmaceutical peptides in biological matrices using HPLC with spectrofluorimetric detection (either intrinsic fluorescence or after a derivatization step; Allen, Stafford, & Nocerini, ; Boppana & Miller‐Stein, ; Chepyala, Tsai, Sun, Lin, & Kuo, ; Hinton et al, ; Lawless, Hopkins, & Anwer, ; Mendoza et al, ). Comparable values of LLOQ (from 5 to 40 ng/mL) have also been reported for peptides quantified in biological fluids with HPLC/MS methods (Gil et al, ; Jiang et al, ; Saraswat et al, ).…”

Section: Resultsmentioning

confidence: 74%

LR12‐peptide quantitation in whole blood by RP‐HPLC and intrinsic fluorescence detection: Validation and pharmacokinetic study

Parent

Boudier

Maincent

et al. 2017

Biomedical Chromatography

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A simple, sensitive, selective and robust HPLC method based on intrinsic fluorescence detection was developed for the quantitation of a dodecapeptide (designated as LR12), inhibitor of Triggering Receptor Expressed on Myeloid cells-1, in rat whole blood. Sample treatment was optimized using protein precipitation and solid-phase extraction. Chromatographic separation was carried out in a gradient mode using a core-shell C column (150 × 4.6 mm, 3.6 μm) with mobile phases of acetonitrile and water containing trifluoroacetic acid at 1.0 mL/min. The method was validated using methodology described by the US Food and Drug Administration guidelines for bioanalytical methods. Linearity was demonstrated within the 50-500 ng/mL range and the lower limit of quantitation was 50 ng/mL. Finally, a preliminary pharmacokinetic study after intraperitoneal injection of LR12 in rats was conducted to evaluate both LR12 monomer and its corresponding disulfide dimer, the main product of degradation. Beyond the fact that this paper describes the first fully validated method for LR12 analysis in blood samples, the approach followed here to optimize pre-analytical steps could be beneficial to develop HPLC and/or MS methods for other pharmaceutical peptides.

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Section: Resultsmentioning

confidence: 74%

LR12‐peptide quantitation in whole blood by RP‐HPLC and intrinsic fluorescence detection: Validation and pharmacokinetic study

Parent

Boudier

Maincent

et al. 2017

Biomedical Chromatography

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“…However, MLIF is a pentapeptide that when administered has a half-life of less than one minute. The reason for its short action time is that it is particularly vulnerable to cleavage by peptidases [ 34 ]. This poses an issue when trying to ascertain the downstream role of this immunomodulatory peptide.…”

Section: Discussionmentioning

confidence: 99%

Monocyte Locomotion Inhibitory Factor Produced byE. histolyticaImproves Motor Recovery and Develops Neuroprotection after Traumatic Injury to the Spinal Cord

Bermeo

Ibarra

García

et al. 2013

BioMed Research International

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Monocyte locomotion inhibitory factor (MLIF) is a pentapeptide produced by Entamoeba histolytica that has a potent anti-inflammatory effect. Either MLIF or phosphate buffered saline (PBS) was administered directly onto the spinal cord (SC) immediately after injury. Motor recovery was evaluated. We also analyzed neuroprotection by quantifying the number of surviving ventral horn motor neurons and the persistence of rubrospinal tract neurons. To evaluate the mechanism through which MLIF improved the outcome of SC injury, we quantified the expression of inducible nitric oxide synthase (iNOS), interleukin-10 (IL-10), and transforming growth factor-β (TGF-β) genes at the site of injury. Finally, the levels of nitric oxide and of lipid peroxidation were also determined in peripheral blood. Results showed that MLIF improved the rate of motor recovery and this correlated with an increased survival of ventral horn and rubrospinal neurons. These beneficial effects were in turn associated with a reduction in iNOS gene products and a significant upregulation of IL-10 and TGF-β expression. In the same way, MLIF reduced the concentration of nitric oxide and the levels of lipid peroxidation in systemic circulation. The present results demonstrate for the first time the neuroprotective effects endowed by MLIF after SC injury.

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Determination of thymopentin in beagle dog blood by liquid chromatography with tandem mass spectrometry and its application to a preclinical pharmacokinetic study

Shi

Yang

Zhou

et al. 2015

J of Separation Science

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The pentapeptide thymopentin (Arg-Lys-Asp-Val-Tyr, RKDVY) corresponds to amino acids 32-36 of the 49 amino acid immunomodulatory polypeptide, thymopoietin, whose biological activity is partially reproduced. Thymopentin is widely used in the clinic and represents a promising target for drug design but bioanalytical and pharmacokinetic data are limited due to its enzymatic instability. This paper reports a rapid and sensitive method based on liquid chromatography with tandem mass spectrometry for the determination of thymopentin in beagle dog blood. To inactivate peptidases and stabilize thymopentin, acetonitrile was added to blood samples immediately after collection followed by addition of stable isotope-labeled thymopentin as internal standard and washing with dichloromethane. Chromatography was carried out on an Ascentis Express Peptide ES-C18 column using gradient elution with methanol and aqueous 0.1% formic acid at a flow rate of 0.6 mL/min. Positive electrospray ionization mass spectrometry with selected reaction monitoring achieved linearity in the range of 1.5-800 ng/mL with good accuracy/precision and minimal matrix effects. The method was successfully applied to a pharmacokinetic study in beagle dogs after intravenous administration of 0.2 mg/kg thymopentin.

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Determination of an unstable pentapeptide, monocyte locomotion inhibitory factor, in dog blood by LC–MS/MS: Application to a pharmacokinetic study

Cited by 10 publications

References 36 publications

LR12‐peptide quantitation in whole blood by RP‐HPLC and intrinsic fluorescence detection: Validation and pharmacokinetic study

LR12‐peptide quantitation in whole blood by RP‐HPLC and intrinsic fluorescence detection: Validation and pharmacokinetic study

Monocyte Locomotion Inhibitory Factor Produced byE. histolyticaImproves Motor Recovery and Develops Neuroprotection after Traumatic Injury to the Spinal Cord

Determination of thymopentin in beagle dog blood by liquid chromatography with tandem mass spectrometry and its application to a preclinical pharmacokinetic study

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