For analysis, HPLC grade solvents, e.g., methanol, acetonitrile, and formic acid (FA), were purchased from Thermo Scientific (Rockford, IL, United States). Water used for analysis was purified using Millipore Synergy® UV Ultrapure Water Purification System (EMD Millipore Corporation, MA, United States). Reference standards, primaquine diphosphate was purchased from Sigma (St Louis, MO, United States). Different metabolites of PQ and internal standards were synthesized at the National Center for Natural Products Research, University of Mississippi. Synthesis of reference standards have been described in the previous article; primaquine-5,6-orthoquinone (POQ) (Potter et al., 2015 (link)), carboxyprimaquine-5,7-orthoquinone (cPOQ) (Khan et al., 2021 (link)), PQ-N-CG (Fasinu et al., 2016 (link)), cPQ (McChesney and Sarangan, 1984 (link)), 4-methylprimaquine-5,6-orthoquinone (4-MePOQ), and deuterated primaquine (d3-PQ) (Khan et al., 2022b (link)). The racemic PQ, PQ enantiomers and placebo capsules used for the clinical study were prepared with complete characterization, purity, and stability, by ElSohly Laboratories, Inc (ELI), Oxford, MS. Each capsule contained the desired specific amount of PQ base as the diphosphate salt. Structure of all analytes are shown in Figure 1 .
Primaquine diphosphate
Manufactured by Merck Group
Sourced in United States
Primaquine diphosphate is a chemical compound used in laboratory research. It is a salt formed by the combination of primaquine and phosphoric acid. Primaquine diphosphate is commonly utilized in scientific investigations, but its specific applications and intended uses are not included in this factual description.
Automatically generated - may contain errors
Lab products found in correlation
Acetonitrile, by Thermo Fisher Scientific (3 mentions)
Milli q system, by Merck Group (2 mentions)
Formic acid, by Merck Group (2 mentions)
Methanol, by Thermo Fisher Scientific (2 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
Quinacrine dihydrochloride, by Merck Group (1 mentions)
Mefloquine hydrochloride, by Merck Group (1 mentions)
Calcimycin, by Merck Group (1 mentions)
Oseltamivir carboxylate, by MedChemExpress (1 mentions)
Amodiaquin dihydrochloride dihydrate, by Merck Group (1 mentions)
Mefloquine, by Merck Group (1 mentions)
5 n n dimethyl amiloride hydrochloride, by Merck Group (1 mentions)
Bafilomycin a, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Capsaicin, by Merck Group (1 mentions)
Chloroquine diphosphate salt, by Merck Group (1 mentions)
Sulfasalazine, by Merck Group (1 mentions)
Thapsigargin, by Merck Group (1 mentions)
N acetyl l cysteine, by Merck Group (1 mentions)
Doxorubicin hydrochloride, by Merck Group (1 mentions)
10 protocols using primaquine diphosphate
1
Quantification of Primaquine Metabolites
2
Antiviral Compound Screening Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Oseltamivir carboxylate was purchased from MedChemexpress CO., Ltd, (Monmouth Junction, NJ). Lysosomotrophic agents amodiaquin dihydrochloride dihydrate, bafilomycin A from Streptomyces griseus, chloroquine diphosphate salt, quinacrine dihydrochloride, quinidine anhydrous, Mefloquine hydrochloride, and primaquine diphosphate were purchased from Sigma-Aldrich (St-Louis, MO). Quinine sulfate was obtained from Fisher (Fair Lawn, NJ). Calcium modulators calcimycin (A23187), capsaicin, 5-(N,N-Dimethyl) amiloride hydrochloride, and verapamil hydrochoride were purchased from Sigma-Aldrich whereas TMB-8 hydrochloride was purchased from Calbiochem (Mississauga, ON). Mefloquine, calcimycin, bafilomycin A, and capsaicin were dissolved in DMSO (Sigma-Aldrich) in order to have a final solvent concentration of less than 0.1%, whereas other compounds were dissolved in culture medium. At this concentration, DMSO showed no apparent toxicity in MDCK cells (less than 1%, data not shown).
3
Purchasing Chemicals for Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primaquine diphosphate (PQ), diethyldithiocarbamate (DDTC), doxorubicin hydrochloride, sulfasalazine (SASP), N-acetyl-L-cysteine (NAC), and thapsigargin (Tg) were purchased from Sigma-Aldrich (Saint Louis, MO, USA).
4
Drug Reference Sample Preparation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Standards and samples of each drug were prepared using the diluents listed in Table 2 . The reference drugs were obtained from Sigma-Aldrich, Castle Hill, Australia (amoxicillin, artesunate, chloroquine diphosphate, doxycycline, primaquine diphosphate, sulphadoxine & pyrimethamine), Sigma Chemicals, Perth, Western Australia (quinine hydrochloride), Sigma Chemicals, St Louis, MO, USA (amodiaquine dihydrochloride dihydrate), AAPIN Chemicals Ltd Abingdon, UK (artemether) and Shaani Sciphar, Biotechnology, Chime Co. Ltd., China (lumefantrine). Standard concentrations were prepared at 1 mg/mL. All stock solutions were stored at 4°C. Chloroquine and amodiaquine were analysed for individual tablet content. Six individual tablets from each batch were weighed, dissolved and analysed individually. For the other drugs, 3–20 tablets from each batch were weighed to obtain the average tablet weight (Table 2 ). The tablets were then crushed to fine powder. Each sample was prepared by dissolving the amount of powder equivalent to the average weight of one tablet. The solutions were sonicated for 10 min × 2 and then equilibrated to room temperature for 30 minutes. A known volume of the clear solution of the first dilution was assayed or further dilutions were done if required to remain within the assay limit (Table 2 ).
5
Synthesis of PEG-Caprolactone Polymers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All chemicals and solvents were of reagent grade and used without additional purification unless otherwise stated. Dichloromethane was acquired from Sigma-Aldrich (St. Louis, MI, USA) and dried using phosphorus pentoxide. Toluene was dried by distillation with sodium and was obtained from Fischer Chemical (Waltham, MA, USA). Poly(ethylene glycol) methyl ether (MePEG, Mn = 5000), ε-caprolactone (CL), and a 2 M solution of hydrogen chloride in diethyl ether were acquired from Sigma-Aldrich (St. Louis, MI, USA). CL was dried with calcium hydride and distilled prior to use. Me-PEG (Mn = 5000) was dried twice by azeotropic distillation in toluene that was distilled off completely. Dimethylformamide (DMF) and acetonitrile (ACN) were acquired from Fisher Chemical (Waltham, MA, USA) and Honeywell (Charlotte, NC, USA), respectively.
Primaquine diphosphate 98% was obtained from Sigma-Aldrich (St. Louis, MI, USA). The gold complexes AuSe and AuS were synthesized and characterized as previously described [14 (link)].
Primaquine diphosphate 98% was obtained from Sigma-Aldrich (St. Louis, MI, USA). The gold complexes AuSe and AuS were synthesized and characterized as previously described [14 (link)].
6
Primaquine Metabolite Synthesis and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Acetonitrile and methanol of HPLC-grade were obtained from Fisher Scientific (Fair Lawn, NJ, USA). Water was purified in a Milli-Q system (Millipore, Bedford, MA, USA). Primaquine diphosphate and formic acid were purchased from Sigma (St Louis, MO, USA). Carboxyprimaquine (cPQ) was prepared using the procedure reported by McChesney and Sarangan [32 (link)]. Primaquine alcohol, 2-hydroxyprimaquine, 3-hydroxyprimaquine, 4-hydroxyprimaquine, 5-hydroxyprimaquine, primaquine-5,6-ortho-quinone, primaquine N-carbamoyl glucuronide, primaquine methyl carbamate, carboxy primaquine lactam, carboxy primaquine methyl ester and primaquine N-acetate were synthesized at National Center for Natural Products Research (NCNPR) and their identity and purity were confirmed by spectral data (IR, NMR and High-Resolution MS) and comparison of their physical data (m.p.) with published values [20 (link), 21 (link)].
7
Primaquine Pharmacokinetics in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Swiss Webster mice were administered 10 mg/kg (primaquine base) of either: (i) primaquine diphosphate (Sigma-Aldrich) solution in water (“Pq,” n = 7 mice per timepoint); (ii) primaquine-loaded galactose-containing nanoemulsion (“NE-Pq,” n = 5 mice per timepoint); or (iii) primaquine-loaded galactose-containing nanochitosan (“NQ-Pq,” n = 7 mice per timepoint). All formulations were given by gavage with a final volume of 200 µL. At different intervals (0.5, 1, 2, 4, 8, and 24 hours), animals were subjected to anesthesia with a mixture of xylazine (10 mg/kg) and ketamine (100 mg/kg). Blood was collected by cardiac puncture, plasma was separated by centrifugation, RBCs were washed three times in saline, and plasma and RBC were stored at −20°C. After the animal was exsanguinated, the liver, lungs, and spleen were collected and frozen in liquid nitrogen until processed for primaquine quantification.
8
Cryopreserved Human Hepatocyte Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cryopreserved primary human hepatocytes were purchased from vendors permitted to sell products derived from human organs procured in the United States by federally designated Organ Procurement Organizations. Vendors included CellzDirect (Lot Hu1434) and Celsis In vitro Technologies (Lot NON). Human hepatocyte culture medium was high glucose Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% (v/v) fetal bovine serum (FBS), 1% (v/v) ITSTM (BD Biosciences), 7 ng/ml glucagon, 40 ng/ml dexamethasone, 15 mM HEPES, and 1% (v/v) penicillin-streptomycin. FIB-3T3 murine embryonic fibroblasts (gift of Howard Green, Harvard Medical School) were cultured at <18 passages in fibroblast medium comprising of DMEM with high glucose, 10% (v/v) bovine serum, and 1% (v/v) penicillin-streptomycin. Polyethylene glycol diacrylate (Mn 3.4 kDa) was purchased from Laysan Bio Inc and used without further purification. Ammonium persulfate (APS), N,N,N′,N′-tetramethylethylenediamine (TEMED), and primaquine diphosphate were obtained from Sigma. primaquine diphosphate was dissolved at 0.5 mg/mL in deionized water, and stored as single-use aliquots at −20 °C.
9
Quantification of Primaquine and Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
primaquine diphosphate ((4-N-(6-methoxyquinolin-8-yl)pentane-1,4-diamine;phosphoric acid; MW = 455); (primaquine; MW = 259)) and 8-aminoquinoline (quinolin-8-amine; MW = 144), internal standard (IS), were from Sigma-Aldrich (St. Louis, MO, USA). 5,6-Orthoquinone primaquine dihydrobromide (8-((5-aminopentan-2-yl)quinoline-5,6-dione dihydrobromide; MW = 259.31) was from Toronto Research Chemicals (Canada). primaquine phosphate was from the Government Pharmaceutical Organization (Thailand). HPLC-grade methanol, acetonitrile, and formic acid were from Sigma-Aldrich (St. Louis, MO, USA). Water was purified in a Milli-Q system (Millipore, Bedford, MA, USA).
10
In vitro Antimalarial Drug Screening
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!