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. 2023 Jun 30;28(2):31-39.
doi: 10.15430/JCP.2023.28.2.31.

Facile Titrimetric Assay of Lysophosphatidic Acid in Human Serum and Plasma for Ovarian Cancer Detection

Nazia Tarannum  1 , Deepak Kumar  1 , Ranu Agrawal  2
Affiliations

Facile Titrimetric Assay of Lysophosphatidic Acid in Human Serum and Plasma for Ovarian Cancer Detection

Nazia Tarannum et al. J Cancer Prev. .

Abstract

Herein, an instrument free facile acid-base titrimetric methodology is reported for lysophosphatidic acid (LPA) measurement in serum and plasma samples for ovarian cancer detection. The concept is based on the titrimetric method in which alkaline solution was titrated with free fatty acid. Free fatty acid is generated due to action of the lysophospholipase to LPA. A phospholipid derivative known as LPA can function as a signaling molecule. A glycerol backbone serves as the foundation for phosphatidic acid, which also has bonds to an unsaturated fatty acid at carbon-1, a hydroxyl group at carbon-2, and a phosphate molecule at carbon-3. Free fatty acid and glycerol-3-phosphate are formed when LPA reacts with lysophospholipase. The formation of free fatty acid depends on the concentration of LPA. The standard graph of known concentrations of LPA, LPA spiked serum and LPA spiked plasma was plotted. The concentration of LPA in unknown serum and plasma were calculated from the standard graph. The limit of detection of LPA in spiked serum and plasma samples via titrimetric assay was calculated as 0.156 μmol/L. A patient's chance of survival may be outweighed by an early diagnosis of ovarian cancer.

Keywords: Acid-base; Assay; Concentration; Lysophosphatidic acid; Titration.

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Conflict of interest statement

CONFLICTS OF INTEREST No potential conflicts of interest were disclosed.

Figures

Figure 1
Figure 1. Survival rate, signaling lipid, and lysophosphatidic acid (LPA) concentrations at different stages of ovarian cancer.
Figure 2
Figure 2. Titrimetric method of acid-base with serial dilution solution and sodium hydroxide (NaOH) standard solution.
Figure 3
Figure 3. Standard graph plot of known lysophosphatidic acid (LPA) concentrations in serial dilutions of (A) PBS, (B) spiked serum, (C) spiked plasma and (D) comparison between the LPA concentration in PBS spiked serum and spiked plasma with volume used in titration.
Figure 4
Figure 4. Chemistry of titrimetric assay.
(A) Step 1 shows change in colour of indicator. (B) Step 2 shows formation of free fatty acid after reaction of LPA with lysophospholipase. (C) Step 3 shows titration of free fatty acid with base in the presence of the indicator. LPA, lysophosphatidic acid; NaOH, sodium hydroxide.
Figure 5
Figure 5. Optimization of the lysophosphatidic acid (LPA) concentration with (A) different incubation times and (B) different amounts of lysophospholipase.
Figure 6
Figure 6. Lysophosphatidic acid (LPA) concentrations in (A) serum sample and (B) plasma sample of different age groups.
Figure 7
Figure 7. Lysophosphatidic acid (LPA) concentrations in serum samples of three volunteers of same age groups (21-30 years).
Figure 8
Figure 8. Effect of thawing and freezing cycles on the concentration of lysophosphatidic acid (LPA) in serum/plasma sample.
Figure 9
Figure 9. Lysophosphatidic acid (LPA) concentrations in serum and plasma of same age group volunteers.
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