Multifunctional reference system for analyte determinations by fluorescence
View Patent ↗The present invention concerns a system and method for the luminescence detection of an analyte in a liquid sample. The system comprises a support on which an analyte-specific substance and a reference substance are located. The analyte-specific substance is able to emit a first luminescence signal on contact with the analyte and the reference substance is able to emit a second luminescence signal which is substantially quenched by contact with the liquid sample.
1. System for the luminescence detection of an analyte in a liquid sample comprising:
(a) a support including an analyte-specific substance which is able to emit a first luminescence signal on contact with the analyte, a reference substance which is able to emit a second luminescence signal which is substantially quenched by contact with the liquid sample, wherein the analyte-specific substance and the reference substance are excitable at essentially the same wavelengths and emit at essentially the same wavelengths, and wherein the analyte-specific substance and the reference substance are fluorophores, and the reference substance is
wherein one of R 2 , R 3 and R 5 is selected from the group consisting of NR 11 R 12 and SO 3 Na;
R 11 and R 12 are selected from the group consisting of hydrogen and C 1 -C 4 alkyl; and
R 1 , R 4 , R 6 , R 7 , R 8 , R 9 and R 10 are selected from the group consisting of hydrogen and a halogen; and
(b) a single detector which is adapted to measure the luminescence signals of the analyte-specific substance and of the reference substance.
2. System of claim 1 , further comprising an evaluation unit formed to determine the amount of sample, the amount of analyte and operational readiness of the support.
3. System of claim 1 , wherein the reference substance has a luminescence signal that is substantially quenched on contact with water.
4. System of claim 1 , wherein the analyte-specific substance is selected from substances whose luminescence signal depends on the pH.
5. System of claim 4 , wherein the analyte-specific substance is
6. System of claim 4 , wherein the analyte-specific substance is
7. System of claim 1 , wherein the reference substance is selected from the group consisting essentially of aminochalcones and chlorochalcones.
8. System of claim 1 , wherein the reference substance is
9. System of claim 1 , wherein the reference substance is
10. System of claim 1 , wherein the reference substance is
11. Method for detecting an analyte in a liquid sample by luminescence comprising:
i) providing a support including an analyte-specific substance which is able to emit a first luminescence signal on contact with the analyte, a reference substance which is able to emit a second luminescence signal which is substantially quenched by contact with the liquid sample, wherein the analyte-specific substance and the reference substance are excitable at essentially the same wavelengths and emit at essentially the same wavelengths, and wherein the analyte-specific substance and the reference substance are fluorophores, and the reference substance is
wherein one of R 2 , R 3 and R 5 is selected from the group consisting of NR 11 R 12 and SO 3 Na;
R 11 and R 12 are selected from the group consisting of hydrogen and C 1 -C 4 alkyl; and
R 1 , R 4 , R 6 , R 7 , R 8 , R 9 and R 10 are selected from the group consisting of hydrogen and a halogen,
ii) contacting the support with a liquid sample which contains an analyte to be detected and
iii) measuring the luminescence signals and detecting the analyte by comparing the luminescence signals.
12. Method of claim 11 , wherein the luminescence signal of the reference substance is measured before bringing the support into contact with the liquid sample.
13. The method of claim 11 further comprising the step of providing an evaluation unit formed to determine the amount of sample.