Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride  (Synonyms: [Ru(DPP)3]Cl2)

Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride ([Ru(dpp)3]²⁺) is an electrochemiluminescent (ECL) probe and oxygen-sensitive sensor. Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride can be used to modify electrode surfaces for the detection of sulfates (S₂O₈^2-) and oxalates, based on electrochemical reactions that generate excited-state species, releasing photons through irreversible redox reactions. Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride utilizes the oxygen quenching of fluorescence mechanism, where fluorescence intensity reflects the metabolic rate of living microorganisms or oxygen levels within cells/tumors, allowing for real-time detection. Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride's main applications include microbial detection, antibacterial activity studies, and tumor microenvironment research^[1][2].

Guidelines (the following is our recommended protocol; this protocol is provided for guidance only and should be modified according to your specific needs).
Tris (4,7-diphenyl-1,10-phenanthroline)ruthenium (II) dichloride ([Ru (DPP)₃]Cl₂) can be applied in the construction of regenerative ECL sensors. As a cathodic detector for persulfate, it can avoid interference from hydrogen evolution, while the anode response to oxalate exhibits a wide linear range.
I. Experimental Procedure for Fluorescence Optical Respirometry (FOR)^[1]:
1. Mix Tris (4,7-diphenyl-1,10-phenanthroline)ruthenium (II) dichloride ([Ru (DPP)₃]Cl₂) with silica gel, embed it in 2% w/w silicone rubber, add 2 drops into a 96-well plate, and incubate in a humidified chamber at 37°C for 2-3 days before use.
2. Culture target microorganisms in vitro (such as Staphylococcus aureus, Escherichia coli, etc.) at 37°C under aerobic conditions for 48 hours, then dilute to 10⁵-10⁶ CFU/mL; prepare test drugs and dilute to the designated concentrations (such as DMSO 2%-40%, medicinal plant extracts 0.08 mg/mL-166 mg/mL).
3. Add 80 μL of drug samples or diluted drug formulations (diluted at ratios of 1:10, 1:100, and 1:1000) into each well of the 96-well plate coated with [Ru (DPP)₃]Cl₂, followed by 80 μL of microbial suspension, then cover the surface with 60 μL mineral oil; use a Tecan Infinite 200PRO or another fluorescence microplate reader to monitor changes in relative fluorescence intensity over time. Perform the experiment in triplicate and record the time required to reach half-maximal fluorescence intensity and the MIC value.
4. Use the serial dilution method and agar plate colony counting method as controls to verify the consistency of FOR detection results, while also evaluating the interference of drug components on the fluorescence signal of [Ru (DPP)₃]Cl₂.

II. In Vitro Experimental Procedure for Intratumoral Oxygen Imaging^[2]:
1. Preparation of nanosensors (Ru-ONPS): Embed [Ru (DPP)₃]Cl₂ dye into F-127 polymer, crosslink with urea and paraformaldehyde, and prepare nanosensors with a particle size of approximately 64 nm.
2. Preparation of cells and co-culture spheroids: Culture tumor cells (such as HCT-8 cells) and normal cells (such as NIH3T3 cells) in vitro and passage them routinely until the logarithmic growth phase; construct tumor cell co-culture spheroids and culture them for designated periods (such as 48 h, 120 h, and 216 h) for oxygen gradient monitoring.
3. Imaging and oxygen level detection: Co-incubate Ru-ONPS with tumor cells or co-culture spheroids, observe cellular uptake of the sensors and intratumoral distribution through confocal microscopy; based on the Stern-Volmer equation, quantitatively detect oxygen levels using changes in the fluorescence signal of the sensors. The detection dynamic range is 0-23 mg/L with a detection limit of 10 μg/mL. Record oxygen content data of cells and co-culture spheroids at different time points (such as 72 h).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

1169.17

C₇₂H₄₈Cl₂N₆Ru

613

455

36309-88-3

Solid

Orange to red

C1(C2=C(C=CC3=C4[N]([Ru+2]567([N]8=C9C%10=C(C(C%11=CC=CC=C%11)=CC=[N]7%10)C=CC9=C(C%12=CC=CC=C%12)C=C8)[N]%13=C%14C%15=C(C(C%16=CC=CC=C%16)=CC=[N]6%15)C=CC%14=C(C%17=CC=CC=C%17)C=C%13)=CC=C3C%18=CC=CC=C%18)C4=[N]5C=C2)=CC=CC=C1.[Cl-].[Cl-]

Room temperature in continental US; may vary elsewhere.

4°C, protect from light, stored under nitrogen

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Hałasa R, et al. Application of tris-(4,7-Diphenyl-1,10 phenanthroline)ruthenium(II) Dichloride to Detection of Microorganisms in Pharmaceutical Products. Pharmaceuticals (Basel). 2023 Jun 8;16(6):856.  [Content Brief]

  [2]. Kumar A, et al. [Ru(dpp)3]Cl2-Embedded Oxygen Nano Polymeric Sensors: A Promising Tool for Monitoring Intracellular and Intratumoral Oxygen Gradients with High Quantum Yield and Long Lifetime. Small. 2024 Apr;20(17):e2307955.  [Content Brief]