DETECTION OF CO2 AND NH3 USING pH SENSITIVE FLUORESCENT NANOSPHERES IMMOBILIZED IN DIFFERENT MATRICE

Aleksandar Szchenyi , Barna Kovcs

University of Pcs, Faculty of Natural Sciences, Institute of Chemistry, Department of General and Physical Chemistry, PCS, Hungary

Introduction: The optical detection of gaseous CO2 and NH3 is often based on a pH sensitive, colorimetric or luminescent indicator. Usually different indicators are selected for the measurement of the two different gases. In case of CO2, indicators having a pKa value greater than 7-8 are commonly used, while molecules with a pKa lower than 6 are proper for sensing ammonia. Having a broad range pH indicator, it is possible to construct sensors for the two gases by using the proper (and different) additives and matrices for CO2 and NH3, respectively. The use of the Fig 1. Fluorescence enhancement mechanism same indicator in two sensors for the two gases means, that we are working close to the two ends on the optical property vs. pH curve: reduced optical signal change, and, hence, a reduced dynamic range should be paid for that. The benefit could be, however, that the same optical design, filter setup, detector and electronics could be used for sensing. In the present work the use of fluorescent, broad range pH sensitive nanospheres immobilized in different matrices for indirect sensing of gaseous CO2 and NH3 are investigated and tested. To eliminate the fluorescence intensity measurement interferences, we have used a dual lifetime referencing (DLR) method. The DLR method uses two fluorescent dyes with overlapping spectroscopic properties, one pH-sensitive, short-lived indicator and a pHinsensitive reference dye with a decay time in the s or ms range. N-allyl-4-piperazinyl-1,8-naphthalimide (APN) have been used as fluorescent pH indicator. Ruthenium(II) tris(diphenylphenanthroline) (Ru(dpp)3) complex has been Fig. 2. APN fluorescence on different pH used as a reference fluorophore. The fluorescence enhancement of the APN in the presence of hydrogen ions, is based on the photon-induced electron transfer mechanism (PET). Experimental: The synthesis of APN was similar to that reported by Niu et al. [2]. The pH sensing principle of APN is shown in Fig 1. and its fluorescence properties on Fig 2. The core of the silica sphere was prepared by dissolving an appropriate amount of Ru(dpp)3 in the tetraethoxysilane (TEOS), after the addition of acid catalyst, ethanol and deionized water, the mixture was stirred to form sol. The sol formation was allowed to proceed for one hour, then the mixture was cooled to 4C and the process was changed from acid catalyzed to base catalyzed by addition of excess of NH4OH. The sol was then added drop wise to mineral oil and stirred vigorously at 200C until silica spheres appeared. The spheres were filtered, washed with ethanol and deionized water. Cocktail for the shell of the sensor was a) b) prepared in to steps. For preventing the leaching of pH sensing dye, APN was covalently bond the to sol-gel precursor vinyltriethoxysilane (VTES) by irradiating their mixture (molar ratio 1:3) with UV lamp (366nm) for 30 minutes. TEOS, ethanol, water and HCl were added, sonicated for 5 minutes and left to form gel for 1 hour. The shell was formed by adding the Ru(dpp) containing spheres to shell forming gel and stirred for one hour at room temperature. The silica spheres was filtered in the centrifuge with 0,22 mm pore diameter filter and washed with ethanol to remove the unreacted components. The resulted particles were dispersed and stored in the ethanol until use. Phase shift measurements were performed with dual-phase lock-in amplifier (DSP830, Stanford Research inc.) in a home made flow through cell. c) d) Optical system consisted of a blue led (430 nm) a band pass filter, Fig. 3. SEM images of the pH sensing nanospheres: Before thermal treatment (a,b) and after the thermal treatment (c,d) bifurcated fiber bundle, and Hamamatsu (H5783-01) PMT with long pass filter (500 nm).
Calibration
0,00 -2,00

Phase shift measurements

-4,00 -6,00 -8,00 -10,00 -12,00 -14,00 -16,00 -18,00 0 1 2 3 4 5 6 7 8 9 10 11 12 13

pH

Fig. 4. Calibration of sensing layer

Results: SEM images of the synthesized nanospheres are shown in the Fig 3.. The main size of the particles was 275 and 380 nm respectively. The thermal treatment of the nanospheres did not effect the morphology, the particles formed a stabile suspension in ethanol. Hydrogen ions strongly enhance the fluorescence intensity of APN, which shows no fluorescence above pH 12. The phase shift of the reference material and the overall sensor particles as a function of the modulation frequency is shown in the Fig 5. It has been found that the difference in the phase shift have a maximal value aroundthe 10 kHz modulation frequency. Fig. 4. shows the obtained calibration curve of the sensing nanospheres in B-R buffer.

Phase shift (deg)

0,0
APN/EtOH

-10,0
Ru(dpp)3/EtOH + Ar

Phase shift (deg)

-20,0 -30,0 -40,0 -50,0 -60,0

Ru(dpp)3+APN at pH 12

Ru(dpp)3+APN at pH 2

Ru(dpp)3+APN in DI water

Ru(dpp)3/TEOS+Ar 200 C 24h

Ru(dpp)3/TEOS+Air dry at 80C

Conclusion: A pH sensing nanospheres has been prepared with covalently bonded pH sensitive dye and co immobilized reference dye. In the pH range 7-11.5 the sensor response has a linear correlation when measuring the overal phase shift. Further investigation is needed for pretreatment and conditioning of the sensing nano spheres to improve their performance in the pH sensing. The final composition of the materials shows promising results for the further development of pH sensor.
[1] I. Klimant, C. Huber, G. Liebsch, G. Neurauter, A. Stangelmayer, O. S. Wolfbeis, New Trends in Fluorescence Spectroscopy, Springer Series on Fluorescence, 2001, 257-274. [2] C.G. Niu, G.M. Zeng, L.X. Chen, G.L. Shen and R.Q. Yu, Analyst 129 (2004), pp. 2024. [3] Torsten Mayr, Ingo Klimant, Otto S. Wolfbeis, Tobias Werner, Analytica Chimica Acta 462 (2002) 110 [4] C.G. Niu, G.M. Zeng, L.X. Chen, G.L. Shen and R.Q. Yu, Analyst 129 (2004), pp. 2024.

-70,0 1000

10000 Modulation Frequency (Hz)

100000

Fig. 5. Phase shift as a functiom of modulation frequency

Acknowledgement: Developing competitiveness of Universities in the South Transdanubian Region (SROP-4.2.1.B-10/2/KONV-2010-0002).