Synthesis · A conductive MOF with bimetallic spontaneously recycled systems as a signal enhancer for the ultrasensitive detection of T-2 toxin using an electrochemical aptasensor

6 structured synthesis routes

Completeness describes how fully the route could be reconstructed from the main article and supporting information.

Partial recipeSource: SI

Route 1: Other

Synthesis of signal probes CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2

Metal precursors2 mg C-Ni1.5Co1.5(HITP)2 and as-prepared AuNPs

Linker precursorsThionine (Thi) and thiolated CP1 DNA capture probe

SolventsUltrapure water and PBS

AdditivesThionine, AuNPs, CP1 DNA

Atmospherenot stated

Temperature4 for overnight CP1 labelling; room temperature not explicitly stated for 4 h loading step

Time4 h stirring for AuNP/Thi loading; overnight CP1 labelling

Work-upThi-Au@C-Ni1.5Co1.5(HITP)2 obtained by centrifugation and dispersed in ultrapure water; final CP1 bioconjugate centrifuged, washed and redispersed in 3 mL PBS.

Show 7 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	C-Ni1.5Co1.5(HITP)2	2 mg	Synthesis of signal probes CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2
Other	as-prepared AuNPs	6 mL then 5 mL	Synthesis of signal probes CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2
Other	Thionine (Thi)	2 mL · 30 mM	Synthesis of signal probes CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2
Solvent	ultrapure water	2 mL suspension; later used for redispersion	Synthesis of signal probes CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2
Other	Thi-Au@C-Ni1.5Co1.5(HITP)2 solution	not stated · 1 mg/mL before second AuNP addition	Synthesis of signal probes CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2
Other	CP1	100 uL · 10 uM	Synthesis of signal probes CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2
Solvent	PBS	1 mL for CP1 reaction; final 3 mL redispersion	Synthesis of signal probes CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2

Complete recipeSource: SI

Route 2: Other

Synthesis of NixCo3-x(HITP)2

Metal precursorsCo(NO3)2.6H2O and Ni(OAc)2.4H2O in DMSO; Co:Ni input ratio 1:1 for Co:Ni; total metal-ion concentration 4.66 mmol/L

Linker precursors15 mg (0.028 mmol) HATP.6HCl in 4.5 mL water

Solvents4.5 mL DMSO for metal salts; 4.5 mL water for HATP.6HCl; 4 mL aqueous NaOAc solution

AdditivesNaOAc aqueous solution, 2 mol/L, 4 mL

Atmospherenot stated

Temperature65

Time2

Work-upBlack powder washed with water and methanol.

ActivationDried under vacuum.

Scalability contextSmall-vial batch: 20 mL glass vial, milligram-scale linker charge.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Co(NO3)2.6H2O	varied by Co:Ni ratio; total metal-ion concentration 4.66 mmol/L in 4.5 mL DMSO · total metal ions 4.66 mmol/L	Synthesis of NixCo3-x(HITP)2
Metal Source	Ni(OAc)2.4H2O	varied by Co:Ni ratio; total metal-ion concentration 4.66 mmol/L in 4.5 mL DMSO · total metal ions 4.66 mmol/L	Synthesis of NixCo3-x(HITP)2
Linker	HATP.6HCl	15 mg (0.028 mmol) in 4.5 mL water	Synthesis of NixCo3-x(HITP)2
Solvent	DMSO	4.5 mL	Synthesis of NixCo3-x(HITP)2
Solvent	water	4.5 mL for linker solution plus 4 mL NaOAc aqueous solution	Synthesis of NixCo3-x(HITP)2
Additive	NaOAc aqueous solution	4 mL · 2 mol/L	Synthesis of NixCo3-x(HITP)2

Partial recipeSource: Both

Route 3: Electrochemical

3 · 2.3 · Scheme 1B

Metal precursors1% (w/v) HAuCl4 for electrodeposited Au film; CP1/Au-Thi-Au@C-Ni1.5Co1.5(HITP)2 bioconjugate

Linker precursorsHP1, S1, S2 DNA probes; BSA blocking layer

Solventsultrapure water, IB, HB, PBS, Exo III buffer

AdditivesBSA, Exo III enzyme, T-2 toxin standards

Atmosphereambient drying; otherwise not stated

Temperature37 for binding/recycling/hybridisation steps; 95 for S1/S2 annealing; 4 for storage

Time0.0083 h annealing at 95 C; 1 h target binding; 2 h S2 immobilisation; 2 h Exo III reaction; 2 h CP1 hybridisation

Substrate orientationglassy carbon electrode polished with 0.05 um alumina slurry

Oxidant / reductantHAuCl4 electrodeposition at -0.2 V vs Ag/AgCl for 30 s

Work-upGently washed and dried naturally after CP1 incubation; electrode rinsed with ultrapure water after each modification step.

Scalability contextAuthors state the electrode construction process is time-consuming and not conducive to practical applications.

Show 8 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	glassy carbon electrode	Not specified	3 · 2.3 · Scheme 1B
Metal Source	HAuCl4	1% (w/v)	3 · 2.3 · Scheme 1B
Other	HP1	10 uL · 10 uM	3 · 2.3 · Scheme 1B
Other	BSA	10 uL · 1% in PBS	3 · 2.3 · Scheme 1B
Other	S1	10 uL · 10 uM	3 · 2.3 · Scheme 1B
Other	S2	10 uL · 10 uM	3 · 2.3 · Scheme 1B
Other	Exo III	1 uL · 200 U	3 · 2.3 · Scheme 1B
Other	CP1 bioconjugate	10 uL	3 · 2.3 · Scheme 1B

Complete recipeSource: SI

Route 4: Other

Synthesis of NixCo3-x(HITP)2

Metal precursorsCo(NO3)2.6H2O and Ni(OAc)2.4H2O in DMSO; Co:Ni input ratio 4:1 for Co:Ni; total metal-ion concentration 4.66 mmol/L

Linker precursors15 mg (0.028 mmol) HATP.6HCl in 4.5 mL water

Solvents4.5 mL DMSO for metal salts; 4.5 mL water for HATP.6HCl; 4 mL aqueous NaOAc solution

AdditivesNaOAc aqueous solution, 2 mol/L, 4 mL

Atmospherenot stated

Temperature65

Time2

Work-upBlack powder washed with water and methanol.

ActivationDried under vacuum.

Scalability contextSmall-vial batch: 20 mL glass vial, milligram-scale linker charge.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Co(NO3)2.6H2O	varied by Co:Ni ratio; total metal-ion concentration 4.66 mmol/L in 4.5 mL DMSO · total metal ions 4.66 mmol/L	Synthesis of NixCo3-x(HITP)2
Metal Source	Ni(OAc)2.4H2O	varied by Co:Ni ratio; total metal-ion concentration 4.66 mmol/L in 4.5 mL DMSO · total metal ions 4.66 mmol/L	Synthesis of NixCo3-x(HITP)2
Linker	HATP.6HCl	15 mg (0.028 mmol) in 4.5 mL water	Synthesis of NixCo3-x(HITP)2
Solvent	DMSO	4.5 mL	Synthesis of NixCo3-x(HITP)2
Solvent	water	4.5 mL for linker solution plus 4 mL NaOAc aqueous solution	Synthesis of NixCo3-x(HITP)2
Additive	NaOAc aqueous solution	4 mL · 2 mol/L	Synthesis of NixCo3-x(HITP)2

Complete recipeSource: SI

Route 5: Other

Synthesis of NixCo3-x(HITP)2

Metal precursorsCo(NO3)2.6H2O and Ni(OAc)2.4H2O in DMSO; Co:Ni input ratio 1:4 for Co:Ni; total metal-ion concentration 4.66 mmol/L

Linker precursors15 mg (0.028 mmol) HATP.6HCl in 4.5 mL water

Solvents4.5 mL DMSO for metal salts; 4.5 mL water for HATP.6HCl; 4 mL aqueous NaOAc solution

AdditivesNaOAc aqueous solution, 2 mol/L, 4 mL

Atmospherenot stated

Temperature65

Time2

Work-upBlack powder washed with water and methanol.

ActivationDried under vacuum.

Scalability contextSmall-vial batch: 20 mL glass vial, milligram-scale linker charge.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Co(NO3)2.6H2O	varied by Co:Ni ratio; total metal-ion concentration 4.66 mmol/L in 4.5 mL DMSO · total metal ions 4.66 mmol/L	Synthesis of NixCo3-x(HITP)2
Metal Source	Ni(OAc)2.4H2O	varied by Co:Ni ratio; total metal-ion concentration 4.66 mmol/L in 4.5 mL DMSO · total metal ions 4.66 mmol/L	Synthesis of NixCo3-x(HITP)2
Linker	HATP.6HCl	15 mg (0.028 mmol) in 4.5 mL water	Synthesis of NixCo3-x(HITP)2
Solvent	DMSO	4.5 mL	Synthesis of NixCo3-x(HITP)2
Solvent	water	4.5 mL for linker solution plus 4 mL NaOAc aqueous solution	Synthesis of NixCo3-x(HITP)2
Additive	NaOAc aqueous solution	4 mL · 2 mol/L	Synthesis of NixCo3-x(HITP)2

Complete recipeSource: SI

Route 6: Other

Synthesis of NixCo3-x(HITP)2

Metal precursorsNi(OAc)2.4H2O in DMSO; no Co added; total metal-ion concentration 4.66 mmol/L

Linker precursors15 mg (0.028 mmol) HATP.6HCl in 4.5 mL water

Solvents4.5 mL DMSO for metal salts; 4.5 mL water for HATP.6HCl; 4 mL aqueous NaOAc solution

AdditivesNaOAc aqueous solution, 2 mol/L, 4 mL

Atmospherenot stated

Temperature65

Time2

Work-upBlack powder washed with water and methanol.

ActivationDried under vacuum.

Scalability contextSmall-vial batch: 20 mL glass vial, milligram-scale linker charge.

Show 5 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Ni(OAc)2.4H2O	varied by Co:Ni ratio; total metal-ion concentration 4.66 mmol/L in 4.5 mL DMSO · total metal ions 4.66 mmol/L	Synthesis of NixCo3-x(HITP)2
Linker	HATP.6HCl	15 mg (0.028 mmol) in 4.5 mL water	Synthesis of NixCo3-x(HITP)2
Solvent	DMSO	4.5 mL	Synthesis of NixCo3-x(HITP)2
Solvent	water	4.5 mL for linker solution plus 4 mL NaOAc aqueous solution	Synthesis of NixCo3-x(HITP)2
Additive	NaOAc aqueous solution	4 mL · 2 mol/L	Synthesis of NixCo3-x(HITP)2