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A fluorescent probe for diacetyl detection.

Li X, Duerkop A, Wolfbeis OS - J Fluoresc (2008)

Bottom Line: A water-soluble fluorescent probe, rhodamine B hydrazide (RBH), was prepared and its properties for recognition of diacetyl were studied.The method employs the reaction of diacetyl with RBH, a colorless and non-fluorescent rhodamine B spiro form derivative to give a pink-colored fluorescent substance.In weakly acidic media, RBH reacts more selectively with diacetyl than with other carbonyls, causing a large increase in fluorescence intensity and thereby providing an easy assay for the determination of diacetyl.

View Article: PubMed Central - PubMed

Affiliation: Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, 93040 Regensburg, Germany.

ABSTRACT
A water-soluble fluorescent probe, rhodamine B hydrazide (RBH), was prepared and its properties for recognition of diacetyl were studied. The method employs the reaction of diacetyl with RBH, a colorless and non-fluorescent rhodamine B spiro form derivative to give a pink-colored fluorescent substance. In weakly acidic media, RBH reacts more selectively with diacetyl than with other carbonyls, causing a large increase in fluorescence intensity and thereby providing an easy assay for the determination of diacetyl.

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Related in: MedlinePlus

Luminescence of supernatants of cancer cell lines before and after spiking with diacetyl. Solid lines (I, III and V) represent unspiked and dashed lines (II, IV and VI) represent spiked samples of SW620 (I and II), LS174 (III and IV) and SW837(V and VI), respectively. A 560 nm (exc.)/590 nm (em) filter combination was used
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Fig6: Luminescence of supernatants of cancer cell lines before and after spiking with diacetyl. Solid lines (I, III and V) represent unspiked and dashed lines (II, IV and VI) represent spiked samples of SW620 (I and II), LS174 (III and IV) and SW837(V and VI), respectively. A 560 nm (exc.)/590 nm (em) filter combination was used

Mentions: The derivatization method was tested in the supernatant of some cell lines. Three different cell lines (SW620, LS174 and SW837) were allowed to undergo their usual metabolism activity in RPMI 1640 Biochrom medium for 2 days. During this period, the medium is enriched with metabolism products and low-molecular weight carbonyl compounds released from the cells. This increases the matrix effect of the supernatant, additionally. RPMI as such is a strong matrix itself because it contains buffer salts and salts for adjustment of ionic strength in g/L concentrations, all 20 amino acids in up to hundreds of mg/L, the vitamins B1, B2, B6, B12; biotin, folic acid and other compounds in lower quantities [26, 27]. After 2 days, the cells were removed by centrifugation and the medium was adjusted to pH 3 with HCl. Unspiked cell medium was tested on its effect on RBH by mixing of 100 μL of cell supernatant, 40 μL of RBH and 60 μL of citric acid-Na2HPO4 buffer of pH 3. From the solid lines in Fig. 6 (curves I, III and V), it is obvious that luminescence increased by a factor between 2 and 5. This points out, that certain amounts of carbonyls were present in the supernatants. The increase in luminescence is completed after about 1 h. This is a shorter time compared to the detection of pure diacetyl and hints to the existence of more reactive carbonyl than diacetyl. We then spiked the cell supernatant with diacetyl (60 μL of supernatant, 40 μL of RBH, 40 μL of 200 μmol/L of diacetyl and 60 μL of buffer) to judge on the capability of RBH to detect diacetyl in the presence of a potentially strongly interfering matrix. On comparing the luminescence of the supernatants spiked with diacetyl with the unspiked samples (I and II, III and IV, V and VI), it is visible that for each cell line, there is a luminescence increase compared to the unspiked supernatant. As the reaction of the spiked samples still takes 3 h to be completed, we deduce that the additional fluorescence increase compared to unspiked samples is due to the presence of diacetyl. This shows that RBH can be used as a fluorescent probe for diacetyl in cell medium as well as in other matrix-containing media in the μmol/L-concentration range, and thus might become a suitable reagent for a rapid screening test for cancerous cells.Fig. 6


A fluorescent probe for diacetyl detection.

Li X, Duerkop A, Wolfbeis OS - J Fluoresc (2008)

Luminescence of supernatants of cancer cell lines before and after spiking with diacetyl. Solid lines (I, III and V) represent unspiked and dashed lines (II, IV and VI) represent spiked samples of SW620 (I and II), LS174 (III and IV) and SW837(V and VI), respectively. A 560 nm (exc.)/590 nm (em) filter combination was used
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2821512&req=5

Fig6: Luminescence of supernatants of cancer cell lines before and after spiking with diacetyl. Solid lines (I, III and V) represent unspiked and dashed lines (II, IV and VI) represent spiked samples of SW620 (I and II), LS174 (III and IV) and SW837(V and VI), respectively. A 560 nm (exc.)/590 nm (em) filter combination was used
Mentions: The derivatization method was tested in the supernatant of some cell lines. Three different cell lines (SW620, LS174 and SW837) were allowed to undergo their usual metabolism activity in RPMI 1640 Biochrom medium for 2 days. During this period, the medium is enriched with metabolism products and low-molecular weight carbonyl compounds released from the cells. This increases the matrix effect of the supernatant, additionally. RPMI as such is a strong matrix itself because it contains buffer salts and salts for adjustment of ionic strength in g/L concentrations, all 20 amino acids in up to hundreds of mg/L, the vitamins B1, B2, B6, B12; biotin, folic acid and other compounds in lower quantities [26, 27]. After 2 days, the cells were removed by centrifugation and the medium was adjusted to pH 3 with HCl. Unspiked cell medium was tested on its effect on RBH by mixing of 100 μL of cell supernatant, 40 μL of RBH and 60 μL of citric acid-Na2HPO4 buffer of pH 3. From the solid lines in Fig. 6 (curves I, III and V), it is obvious that luminescence increased by a factor between 2 and 5. This points out, that certain amounts of carbonyls were present in the supernatants. The increase in luminescence is completed after about 1 h. This is a shorter time compared to the detection of pure diacetyl and hints to the existence of more reactive carbonyl than diacetyl. We then spiked the cell supernatant with diacetyl (60 μL of supernatant, 40 μL of RBH, 40 μL of 200 μmol/L of diacetyl and 60 μL of buffer) to judge on the capability of RBH to detect diacetyl in the presence of a potentially strongly interfering matrix. On comparing the luminescence of the supernatants spiked with diacetyl with the unspiked samples (I and II, III and IV, V and VI), it is visible that for each cell line, there is a luminescence increase compared to the unspiked supernatant. As the reaction of the spiked samples still takes 3 h to be completed, we deduce that the additional fluorescence increase compared to unspiked samples is due to the presence of diacetyl. This shows that RBH can be used as a fluorescent probe for diacetyl in cell medium as well as in other matrix-containing media in the μmol/L-concentration range, and thus might become a suitable reagent for a rapid screening test for cancerous cells.Fig. 6

Bottom Line: A water-soluble fluorescent probe, rhodamine B hydrazide (RBH), was prepared and its properties for recognition of diacetyl were studied.The method employs the reaction of diacetyl with RBH, a colorless and non-fluorescent rhodamine B spiro form derivative to give a pink-colored fluorescent substance.In weakly acidic media, RBH reacts more selectively with diacetyl than with other carbonyls, causing a large increase in fluorescence intensity and thereby providing an easy assay for the determination of diacetyl.

View Article: PubMed Central - PubMed

Affiliation: Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, 93040 Regensburg, Germany.

ABSTRACT
A water-soluble fluorescent probe, rhodamine B hydrazide (RBH), was prepared and its properties for recognition of diacetyl were studied. The method employs the reaction of diacetyl with RBH, a colorless and non-fluorescent rhodamine B spiro form derivative to give a pink-colored fluorescent substance. In weakly acidic media, RBH reacts more selectively with diacetyl than with other carbonyls, causing a large increase in fluorescence intensity and thereby providing an easy assay for the determination of diacetyl.

Show MeSH
Related in: MedlinePlus