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. 2015 Aug 25;10 Spec Iss(Spec Iss):93-9.
doi: 10.2147/IJN.S88378. eCollection 2015.

Measurement of creatinine in human plasma using a functional porous polymer structure sensing motif

Sitansu Sekhar Nanda  1 , Seong Soo A An  1 , Dong Kee Yi  2
Affiliations

Measurement of creatinine in human plasma using a functional porous polymer structure sensing motif

Sitansu Sekhar Nanda et al. Int J Nanomedicine. .

Abstract

In this study, a new method for detecting creatinine was developed. This novel sensor comprised of two ionic liquids, poly-lactic-co-glycolic acid (PLGA) and 1-butyl-3-methylimidazolium (BMIM) chloride, in the presence of 2',7'-dichlorofluorescein diacetate (DCFH-DA). PLGA and BMIM chloride formed a functional porous polymer structure (FPPS)-like structure. Creatinine within the FPPS rapidly hydrolyzed and released OH(-), which in turn converted DCFH-DA to DCFH, developing an intense green color or green fluorescence. The conversion of DCFH to DCF(+) resulted in swelling of FPPS and increased solubility. This DCF(+)-based sensor could detect creatinine levels with detection limit of 5 µM and also measure the creatinine in blood. This novel method could be used in diagnostic applications for monitoring individuals with renal dysfunction.

Keywords: FPPS; PLGA; blood; creatinine; diagnosis.

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Figures

Figure 1
Figure 1
PLGA with functional porous polymer structure. Note: (A) The SEM image of PLGA (control) and (B) pores in PLGA completely cross-linked with BMIM chloride. Abbreviations: BMIM, 1-butyl-3-methylimidazolium; PLGA, poly-lactic-co-glycolic acid; SEM, scanning electron microscopy.
Figure 2
Figure 2
This equation shows that when PLGA mixed with ionic liquid (BMIM chloride) in the presence of dichloromethane, PLGA slurry is formed. Note: After washing with ethanol, it confirmed ester linkage with pore. Abbreviations: BMIM, 1-butyl-3-methylimidazolium; PLGA, poly-lactic-co-glycolic acid.
Figure 3
Figure 3
The average size of the pore. Note: It was 18.214 µm on the X-axis, 17.946 µm on the Y-axis, and the diameter of the pore was 25.570 µm.
Figure 4
Figure 4
FPPS and DCFH-DA were used to screen different biomolecules such as glucose, lactose, and urea along with creatinine. Note: Only creatinine showed green fluorescence under UV-visible light (365 nm) and green color in normal white light. Abbreviations: DCFH-DA, 2′,7′-dichlorofluorescein diacetate; FPPS, functional porous polymer structure; UV, ultraviolet.
Figure 5
Figure 5
FPPS with DCFH-DA showed no fluorescence intensity. Note: Addition of 50 µM of creatinine resulted in fluorescence intensity at 530 nm. Abbreviations: au, absorption unit; DCFH-DA, 2′,7′-dichlorofluorescein diacetate; FPPS, functional porous polymer structure; PLGA, poly-lactic-co-glycolic acid.
Figure 6
Figure 6
The creatinine concentration was higher in blood samples from diabetic patients and showed more fluorescence intensity compared with blood from normal individuals. Abbreviation: au, absorption unit.
Figure 7
Figure 7
As the concentration of creatinine increased, the fluorescence intensity (at 530 nm) of the samples also increased. Note: A sharp increase in the emission peak was observed as the concentration of creatinine increased. Abbreviation: au, absorption unit.
Figure 8
Figure 8
Creatinine levels in five blood samples from healthy individuals, five from patients with Alzheimer’s, and five from diabetic patients. Notes: (A) Sample contained creatinine in normal eye with Alzheimer’s blood, Diabetes blood and Normal blood. (B) Sample contained creatinine under UV light at 365 nm with Alzheimer’s blood, Diabetes blood and Normal blood. They also produced green color in normal white light. The color intensity in the blood samples from diabetic patients was higher compared to other blood samples.
Figure 9
Figure 9
The optical density of blood sample. Notes: Creatinine levels in five blood samples from healthy individuals, five from patients with Alzheimer’s, and five from diabetic patients. *P<0.001 by one-way ANOVA followed by Tukey’s test.
Figure 10
Figure 10
Schematic diagram of measuring creatinine using FPPS. Notes: The schematic diagram showed pore formation by PLGA when BMIM chloride was added. As the concentration of DCF+ increased, the green color of the solution intensifies. Abbreviations: BMIM, 1-butyl-3-methylimidazolium; DCFH-DA, 2′,7′-dichlorofluorescein diacetate; FPPS, functional porous polymer structure; PLGA, poly-lactic-co-glycolic acid.
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