Fluorimetric Detection of Insulin in the Presence of Eu(III) - {Pyridine - 2,6 - Dicarboxylate} Tris Complex

Elmas GÖK; Serdar ATEŞ

Makale detayları

Fluorimetric Detection of Insulin in the Presence of Eu(III) - {Pyridine - 2,6 - Dicarboxylate} Tris Complex

Yazarlar:

Elmas GÖK | Serdar ATEŞ

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Chemistry 2001 Vol.25 Issue 1, pp.81-91

Belge Tipi:

Makale

Yayın Tarihi:

12/2000

Belge Dili:

Türkçe

Document ID (Org No.):

2000730188

Sıklık:

İki Aylık

eISSN:

1303-6130

pISSN:

1300-0527

Özet:

 

Bovine insulin solutions (pH=9.3) have maximum absorption at 278 \pm  2 nm and an intrinsic emission at 305 nm when excited at 282 nm.  The  relative fluorescence intensities show linear dependence on its  concentration: 10 mg/mL< [Ins] < 200 mg/mL. When  Eu(PDA)33- tris complex is added to these solutions, it has a  hyperchromic effect at 278 nm absorption band of insulin, sensitizing  the emission intensities of central Eu3+ metal ion of the complex  at 590 and 615 nm, and simultaneously quenching the emission intensity  of hormone at 305 nm.  Stern-Volmer plots show that a mechanism of  bimolecular quenching at 305 nm and sensitization at 615 nm are valid  up to a mole ratio, R = [Eu(PDA)33-]/[Ins] < 2.0.  An  intramolecular rather than an intermolecular energy transfer is  proposed.  An apparent binding constant, log Kapp=4.70 \pm  0.13, is calculated for Ins-[Eu(PDA)33-]2 type product,  the presence of which may offer a new luminescence technique as a  diagnostic tool and an alternative to radio-iodinated (131I -)  insulin.                   A simple, rapid and accurate quantitation of insulin is proposed by  using a fixed concentration of Eu(PDA)33-, and measuring its  initial F0 at lexc/ lem=282/615 nm and the  difference, DF after sensitization when \sim 100 microliters of  insulin sample is added.  The coefficient of variation (CV), the  relative error and minimum detectable amount of bovine insulin hormone  are found to be 3.0%, 1.2% and 7.3 \pm 0.2 mg /mL  respectively.

 

Bovine insulin solutions (pH=9.3) have maximum absorption at 278 \pm  2 nm and an intrinsic emission at 305 nm when excited at 282 nm.  The  relative fluorescence intensities show linear dependence on its  concentration: 10 mg/mL< [Ins] < 200 mg/mL. When  Eu(PDA)33- tris complex is added to these solutions, it has a  hyperchromic effect at 278 nm absorption band of insulin, sensitizing  the emission intensities of central Eu3+ metal ion of the complex  at 590 and 615 nm, and simultaneously quenching the emission intensity  of hormone at 305 nm.  Stern-Volmer plots show that a mechanism of  bimolecular quenching at 305 nm and sensitization at 615 nm are valid  up to a mole ratio, R = [Eu(PDA)33-]/[Ins] < 2.0.  An  intramolecular rather than an intermolecular energy transfer is  proposed.  An apparent binding constant, log Kapp=4.70 \pm  0.13, is calculated for Ins-[Eu(PDA)33-]2 type product,  the presence of which may offer a new luminescence technique as a  diagnostic tool and an alternative to radio-iodinated (131I -)  insulin.                   A simple, rapid and accurate quantitation of insulin is proposed by  using a fixed concentration of Eu(PDA)33-, and measuring its  initial F0 at lexc/ lem=282/615 nm and the  difference, DF after sensitization when \sim 100 microliters of  insulin sample is added.  The coefficient of variation (CV), the  relative error and minimum detectable amount of bovine insulin hormone  are found to be 3.0%, 1.2% and 7.3 \pm 0.2 mg /mL  respectively.

Makale detayları

Fluorimetric Detection of Insulin in the Presence of Eu(III) - {Pyridine - 2,6 - Dicarboxylate} Tris Complex

Yazarlar: Elmas GÖK | Serdar ATEŞ

Yayıncı: TUBITAK

Dergi: Turkish Journal of Chemistry 2001 Vol.25 Issue 1, pp.81-91

Belge Tipi: Makale

Yayın Tarihi: 12/2000

Belge Dili: Türkçe

Document ID (Org No.): 2000730188

Sıklık: İki Aylık

eISSN: 1303-6130

pISSN: 1300-0527

Sayı

Yazarlar:

Elmas GÖK | Serdar ATEŞ

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Chemistry 2001 Vol.25 Issue 1, pp.81-91

Belge Tipi:

Makale

Yayın Tarihi:

12/2000

Belge Dili:

Türkçe

Document ID (Org No.):

2000730188

Sıklık:

İki Aylık

eISSN:

1303-6130

pISSN:

1300-0527