A New Substrate for Alkaline Phosphatase Based on Quercetin ...
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A New Substrate for Alkaline Phosphatase Based on Quercetin ...
The purpose of this section is to provide additional information of interest to the reader: The information include, a pictogram showing changes of QPP solution ...
A New Substrate for Alkaline Phosphatase Based on Quercetin Pentaphosphate Samuel K. Mwilu, Veronica A. Okello, Francis Osonga, Seth Miller, Omowunmi A. Sadik* Department of Chemistry Center for Advanced Sensors & Environmental Monitoring (CASE) State University of New York-Binghamton P. O. Box 6000 Binghamton, NY 13902
United States Environmental Protection Agency /Office of Research & Development National Environmental Research P.O. Box 93478 Las Vegas, NV 89193-3478
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The purpose of this section is to provide additional information of interest to the reader: The information include, a pictogram showing changes of QPP solution before and after reacting with AP, changes in QPP absorbance and Rabbit anti-BG conjugated with ALP response of QPP substrate.
Scheme 1S: Synthetic procedures for QPP according to reference # 14
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A
B
C
D
Figure S1. Pictogram showing the changes in color of a QPP solution after interaction with ALP. (A) before addition of ALP (B) 5 min after adding ALP to one vial (C) 10 min (D) 30 min
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Figure S2. CV obtained for the oxidation of quercetin adsorbed on gold working electrode scan rate 50 mV/s; 1µA/V. All experiments were carried out in phosphate buffer pH 7.03; potential range -200 – 600 mV vs Ag/AgCl. The blank contains phosphate buffer only.
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0.20
y=0.0102x+0.026 R2=0.9848
0.18 0.16
Absorbance
0.14 0.12 0.10 0.08 0.06 0.04 0.02 0.00 0
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[ALP] U/L Figure S3. Change in absorbance (Abs410nm- Abs324nm) in the presence of different concentrations of ALP. The absorbance readings were taken 18 minutes after incubation with ALP enzyme in DEA buffer and corrected for blank.
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0.04
Response
0.03 0.02 0.01 0.00 0
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[BG spores] x 1000 spores/ml Figure S4. Rabbit anti-BG conjugated with ALP response of QPP substrate. Absorbance (410-
324nm) response with increasing concentrations of the BG spores when QPP was used as substrates. A limit of detection of 5998 spores/ml was achieved.R2=0.989
