Development and validation of an LC–MS/MS method to quantify kynurenic acid in human plasma: supplementary data

Background: Monitoring levels of endogenous biomarkers has become an alternative approach to assess transporter-mediated drug–drug interactions in clinical trials. Among the biomarkers of interest, kynurenic acid is effective for the human organic anion transporters OAT1 and OAT3. Here, a simple and robust bioanalytical method was developed using LC–MS/MS to quantify kynurenic acid in human plasma. Results: This method achieved a LLOQ of 10 nm with acceptable signal-to-noise ratio (S/N >5). In addition, an interfering agent, tryptophan, was identified and separated chromatographically. A full method validation was performed in the spirit of GLP. Conclusion: This method can serve as a tool readily available to assess potential drug–drug interactions mediated by inhibition of OAT1 and OAT3 activities.</p

The relationship between sand coverage and interfacial shear strength.

<p>The relationship between sand coverage and interfacial shear strength.</p

The effect curve of each factor based on the orthogonal experiment result.

<p>The effect curve of each factor based on the orthogonal experiment result.</p

The relationship between sand grain size and interfacial shear strength.

<p>The relationship between sand grain size and interfacial shear strength.</p

The relationship between adhesive curing time and interfacial shear strength.

<p>The relationship between adhesive curing time and interfacial shear strength.</p

miR-224-5p expression level was downregulated in uveal melanoma cell lines and samples and inversely correlated with FTH1P3.

<p>(A) The expression level of miR-224-5p in the uveal melanoma cell lines (C918, MUM-2B, OCM-1A and MUM-2C) and melanocyte cell line (D78) was determined by qRT-PCR. (B) The miR-224-5p expression was lower in the uveal melanoma samples than in the no-tumor samples. (C) The expression of miR-224-5p in the uveal melanoma tissues was inversely correlated with FTH1P3 expression. ***p<0.001.</p

Elevated expression of FTH1P3 increased the uveal melanoma cell proliferation and migration.

<p>(A) The expression of FTH1P3 in the uveal melanoma cell line MUM-2B treated with pcDNA-FTH1P3 was detected with qRT-PCR. (B) The cell proliferation was meaured by CCK-8 assay. Ectopic expression of FTH1P3 promoted the MUM-2B cell proliferation. (C) Elevated expression of FTH1P3 increased the MUM-2B cell cycle. (D) Wound healing assay was performed to determine the cell migration. (E) The relative ratio of wound closure per field was shown. *p<0.05, **p<0.01 and ***p<0.001.</p

The diagram of oil and gas well.

<p>The diagram of oil and gas well.</p

Levels for orthogonal experiment.

<p>Levels for orthogonal experiment.</p

The relationship between adhesive curing time and bonding strength.

<p>The relationship between adhesive curing time and bonding strength.</p