Animals were administered cefepime hydrochloride (Hospira, Lake Forest, IL [lot no. 144G005]) for injection. Artificial CSF (Tocris Bioscience, Bristol, UK) and normal saline (Abbott [lot no. C950212]) were used as in the sampling methods described below. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) standard curves were generated using commercially obtained cefepime hydrochloride (USP) (Chem-Impex International, Inc., Wood Dale, IL) with a purity of greater than 99.5%. Ceftazidime pentahydrate (Acros Organics, NJ) was used as an internal standard for the cefepime quantification. Formic acid was obtained from Thermo Fisher Scientific (Waltham, MA). Acetonitrile (BDH [lot no. 16I011254]) and methanol (BDH [lot no. 16L204024]) were obtained from VWR Analytical (Chicago, IL). Milli-Q water was obtained from Midwestern University via an Aqua Solutions water purification dispensatory. Frozen, nonmedicated, nonimmunized, pooled Sprague-Dawley rat plasma (anticoagulated with disodium EDTA) and CSF were used for calibration of standard curves (BioreclamationIVT, Westbury, NY).
Ceftazidime pentahydrate
Manufactured by Thermo Fisher Scientific
Sourced in Germany, Belgium
Ceftazidime pentahydrate is a laboratory reagent used in the preparation of microbiology media and for the detection and identification of bacteria in clinical and research settings. It is a broad-spectrum cephalosporin antibiotic that inhibits bacterial cell wall synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Formic acid, by Thermo Fisher Scientific (1 mentions)
Artificial csf, by Bio-Techne (1 mentions)
Kanamycin sulfate, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Carl Roth (1 mentions)
Trovafloxacin mesylate, by Merck Group (1 mentions)
Gentamycin sulfate, by Carl Roth (1 mentions)
Tetracycline hydrochloride, by Carl Roth (1 mentions)
Soluble starch, by Merck Group (1 mentions)
Trichloroacetic acid, by Merck Group (1 mentions)
Folin ciocalteu reagent, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Rutin, by Merck Group (1 mentions)
Casein, by Merck Group (1 mentions)
Amyloglucosidase, by Merck Group (1 mentions)
Sodium acetate trihydrate, by Chempur (1 mentions)
Viscozyme l, by Merck Group (1 mentions)
Hexamethylenetetramine, by Carl Roth (1 mentions)
Cellulase, by Merck Group (1 mentions)
4 protocols using ceftazidime pentahydrate
1
Cefepime Quantification in Biological Fluids
2
Optimizing Rapid CAZ/AVI NP Test Parameters
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On the basis of our previous experience developing several rapid diagnostic NP tests, we set and compared different parameters to determine the optimal conditions of the Rapid CAZ/AVI NP test by using 2 CAZ/AVI–susceptible isolates (1 E. coli ATCC 25922 and 1 KPC3-producing K. pneumoniae 3074) as negative controls and 2 CAZ/AVI–resistant isolates (1 NDM-5-producing E. coli 3031 and 1 KPC-41-producing K. pneumoniae 3007) as positive controls. Those parameters included bacterial inoculum, 98% ceftazidime pentahydrate (Acros Organics, Thermofisher Scientific) concentrations, avibactam sodium hydrate (MedChem Express, distributed by Lucerna-Chem, https://lucerna-chem.ch ) concentrations, and incubation times with and without shaking. After comparison of the results with different parameters, all experiments were performed in triplicate by 2 persons using the optimal protocol obtained, as described below.
3
Determination of Antibiotic Susceptibility
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Stock solutions of kanamycin sulfate (Fisher
Chemical, U.K.), cefotaxime sodium salt (Roth, Germany), gentamycin
sulfate (Toku-E), spectinomycin dihydrochloride pentahydrate (Roth,
Germany), tetracycline hydrochloride (Roth, Germany), doxycycline
hyclate (Roth, Germany), ceftazidime pentahydrate (Acros Organics,
Belgium), amikacin sulfate (Toku-E), tobramycin sulfate (Toku-E),
fosfomycin sulfate (Toku-E), imipenem (Toku-E), and meropenem (Toku-E)
were dissolved in distilled water. Chloramphenicol (Toku-E) and levofloxacin
(Toku-E) were dissolved in 50% ethanol. Norfloxacin (Toku-E) was dissolved
in 5% acetic acid (POCH, Poland). Ciprofloxacin (Toku-E) was dissolved
in 0.1 M NaOH (POCH, Poland). Trovafloxacin mesylate (Sigma, Germany)
and nitrofurantoin (Toku-E) were dissolved in DMSO (Roth, Germany).
We determined the minimal inhibitory concentrations (MICs) of testing
antibiotics by the standard broth dilution method. After overnight
incubation on 96-well plates, we estimated MIC values: 1.5 μg/mL
kanamycin, 0.64 μg/mL cefotaxime, 0.75 μg/mL gentamycin,
3 μg/mL spectinomycin, 1.5 μg/mL tetracycline, 0.5 μg/mL
doxycycline, 1 μg/mL chloramphenicol, 0.02 μg/mL norfloxacin,
0.006 μg/mL ciprofloxacin, 0.004 μg/mL levofloxacin, 4
μg/mL amikacin, 4 μg/mL fosfomycin, 0.004 μg/mL
trovafloxacin, 1 μg/mL ceftazidime, 1.5 μg/mL tobramycin,
0.75 μg/mL imipenem, 0.1 μg/mL meropenem, and 3 μg/mL
nitrofurantoin.
Chemical, U.K.), cefotaxime sodium salt (Roth, Germany), gentamycin
sulfate (Toku-E), spectinomycin dihydrochloride pentahydrate (Roth,
Germany), tetracycline hydrochloride (Roth, Germany), doxycycline
hyclate (Roth, Germany), ceftazidime pentahydrate (Acros Organics,
Belgium), amikacin sulfate (Toku-E), tobramycin sulfate (Toku-E),
fosfomycin sulfate (Toku-E), imipenem (Toku-E), and meropenem (Toku-E)
were dissolved in distilled water. Chloramphenicol (Toku-E) and levofloxacin
(Toku-E) were dissolved in 50% ethanol. Norfloxacin (Toku-E) was dissolved
in 5% acetic acid (POCH, Poland). Ciprofloxacin (Toku-E) was dissolved
in 0.1 M NaOH (POCH, Poland). Trovafloxacin mesylate (Sigma, Germany)
and nitrofurantoin (Toku-E) were dissolved in DMSO (Roth, Germany).
We determined the minimal inhibitory concentrations (MICs) of testing
antibiotics by the standard broth dilution method. After overnight
incubation on 96-well plates, we estimated MIC values: 1.5 μg/mL
kanamycin, 0.64 μg/mL cefotaxime, 0.75 μg/mL gentamycin,
3 μg/mL spectinomycin, 1.5 μg/mL tetracycline, 0.5 μg/mL
doxycycline, 1 μg/mL chloramphenicol, 0.02 μg/mL norfloxacin,
0.006 μg/mL ciprofloxacin, 0.004 μg/mL levofloxacin, 4
μg/mL amikacin, 4 μg/mL fosfomycin, 0.004 μg/mL
trovafloxacin, 1 μg/mL ceftazidime, 1.5 μg/mL tobramycin,
0.75 μg/mL imipenem, 0.1 μg/mL meropenem, and 3 μg/mL
nitrofurantoin.
4
Characterization of Enzymatic Activities
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hexamethylenetetramine (≥99%), aluminum chloride (≥98%), agar (Agar-Agar), and LB broth were obtained from Carl Roth Gmbh & Co Kg (Karlsruhe, Germany). The 2,2-diphenyl-1-picrylhydrazyl (DPPH) (99%), methanol (≥99.9%), rutin (95%), Viscozyme® L, cellulase from Aspergillus niger, Clara-diastase, lipase from Aspergillus oryzae, protease from Bacillus species, amyloglucosidase from Aspergillus niger, Tween 20 broth, trichloroacetic acid (≥99%), starch (soluble) and 3,5-dinitrosalicylic acid (DNS) were obtained from Sigma-Aldrich Corporation (Taufkirchen, Germany). Folin–Ciocalteu reagent, casein and acetonitrile (≥99.8%) were supplied by Merck (Darmstadt, Germany). Sodium carbonate, calcium carbonate, and acetic acid (99.9%) were bought from Reachem S.r.o. (Bratislava, Slovakia). Sodium acetate (trihydrate) (99%) was obtained from ChemPur (Piekary Śląskie, Poland). Cellulose and ceftazidime pentahydrate (98%) were obtained from Acros Organics (Geel, Belgium). Bidistilled water was prepared by means of the distillation apparatus Thermo Scientific (Fremont, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!