The established method described by Agarwal et al. was used [69 (link)]. Briefly, this method uses a microplate assay based on the reaction of p-dimethylaminocinnamaldehyde (Sigma-Aldrich, Taufkirchen, Germany) with PAH. The results thus obtained are in good agreement with HPLC-based methods. For a more detailed description of the quantitation method, we refer to the in-depth explanations in our previous study [32 (link)].
P dimethylaminocinnamaldehyde
Manufactured by Merck Group
Sourced in Germany, United States
P-dimethylaminocinnamaldehyde is a chemical compound used as a reagent in analytical chemistry procedures. It is a yellow crystalline solid that is soluble in organic solvents. The compound is commonly used as a test for the detection and identification of certain organic compounds.
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Lab products found in correlation
Gallic acid, by Merck Group (2 mentions)
Catechin, by Merck Group (2 mentions)
Sodium chloride (nacl), by Merck Group (2 mentions)
Sodium carbonate, by Merck Group (2 mentions)
Hydrochloric acid (hcl), by Chron Chemicals (1 mentions)
Sodium hydroxide (naoh), by Chron Chemicals (1 mentions)
Acetone, by Chron Chemicals (1 mentions)
Acetic acid, by Chron Chemicals (1 mentions)
Oxalic acid, by Sangon (1 mentions)
Aluminum trichloride, by Chron Chemicals (1 mentions)
Lutein, by Solarbio (1 mentions)
Authentic ascorbic acid, by Solarbio (1 mentions)
Sulfatase, by Merck Group (1 mentions)
Deae sephadex a 25, by Merck Group (1 mentions)
Procyanidin b2, by Merck Group (1 mentions)
2 4 6 tris 2 pyridyl s triazine, by Merck Group (1 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid, by Sangon (1 mentions)
β carotene, by Solarbio (1 mentions)
Ortho nitrophenyl β d galactopyranoside, by Merck Group (1 mentions)
Quercetin, by Solarbio (1 mentions)
8 protocols using p dimethylaminocinnamaldehyde
1
Microplate Assay for PAH Quantitation
2
Microplate Assay for PAH Quantification
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Essentially, the method described by Agarwal et al. was applied (Agarwal 2002 (link)). This microplate assay based on the reaction of p-dimethylaminocinnamaldehyde (Sigma–Aldrich, Taufkirchen, Germany) with PAH gives results that are in very good agreement with HPLC-based methods. For plasma, the precision was 2.5 and 3.2%, and the accuracy was 98 and 100% for solutions containing 12 μg or 24 μg PAH per mL. For urine, the precision was 2.8 and 4.1% and the accuracy was 100 and 97% for solutions containing 30 μg or 60 μg PAH per mL.
3
Phytochemical Analysis of Plant Extracts
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Analytical grade of sodium hypochlorite, oxalic acid, Folin–Ciocalteu, sodium carbonate, gallic acid, sodium salt trihydrate, and 2,2-azinobis(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) were obtained from Sangon Biotech Co., Ltd. (shanghai, China). Analytical grade of ethanol, acetone, acetic acid, hydrochloric acid, aluminum trichloride, ferrous sulfate (FeSO4·7H2O), pyridine, and sodium hydroxide were purchased from Chengdu Kelong Chemical Co., Ltd. (Chengdu, China). The standards of chlorophylls (a and b), carotenoids (neoxanthin, violaxanthin, lutein, and β-carotene), soluble sugars (glucose, fructose, and sucrose), authentic ascorbic acid, and quercetin were obtained from Solarbio Science & Technology Co., Ltd. (Beijing, China). High-performance liquid chromatography (HPLC) grade of p-dimethylaminocinnamaldehyde, 2,4,6-tris(2-pyridyl)-s-triazine, ortho-nitrophenyl β-d -galactopyranoside, sulfatase, and DEAE-sephadex A-25, as well as Procyanidin B2 standards were purchased from Sigma Chemical Co. (Saint Louis, USA). HPLC grade of isopropanol, acetonitrile, and methyl alcohol were purchased from Tedia Company, Inc. (Fairfield, USA).
4
PAH Quantitation via Microplate Assay
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For quantitation of PAH, we utilized the established method as described by Agarwal et al. [79 (link)]; briefly, this refers to the use of a microplate assay relying on the reaction of p-dimethylaminocinnamaldehyde (Sigma-Aldrich, Taufkirchen, Germany) with PAH, which provides results that conform to a large degree with HPLC-based methods. A comprehensive description of the method mentioned above is supplied in our previous study [49 (link)].
5
Antimicrobial Textile Evaluation
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Twisted multifilament cotton (TMC) thread
(Simco) and piping white glazed cotton (PWGC) thread (Simco) were
purchased from a local craft store. Sodium chloride (NaCl), hydrogen
peroxide (H2O2), sodium carbonate (Na2CO3), acetic acid, glycerol, urea, and glucose were procured
from Merck, India. Hydrochloric acid (HCl), n-heptane,
and potassium hydroxide were obtained from Loba Chemie and SRL, India.l -proline β-naphthylamide (PRO) and p-dimethylaminocinnamaldehyde (DCA) were purchased from Sigma-Aldrich,
India, and Loba Chemie, India. Calcium hydroxide, bovine serum albumin
(BSA), and lactic acid were procured from Titan Biotech, HiMedia,
and Finar Limited, India. Sanitary napkins and tampons (Bella) were
procured from a local pharmacy. C. albicans (ATCC 24433) was taken from the culture collection at Mycology Laboratory,
Department of Microbiology, Kasturba Medical College, Manipal.
(Simco) and piping white glazed cotton (PWGC) thread (Simco) were
purchased from a local craft store. Sodium chloride (NaCl), hydrogen
peroxide (H2O2), sodium carbonate (Na2CO3), acetic acid, glycerol, urea, and glucose were procured
from Merck, India. Hydrochloric acid (HCl), n-heptane,
and potassium hydroxide were obtained from Loba Chemie and SRL, India.
India, and Loba Chemie, India. Calcium hydroxide, bovine serum albumin
(BSA), and lactic acid were procured from Titan Biotech, HiMedia,
and Finar Limited, India. Sanitary napkins and tampons (Bella) were
procured from a local pharmacy. C. albicans (ATCC 24433) was taken from the culture collection at Mycology Laboratory,
Department of Microbiology, Kasturba Medical College, Manipal.
6
Quantifying Phenolic Compounds Analysis
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Fluorescein, Folin–Ciocalteu reagent, gallic acid, p-dimethylaminocinnamaldehyde (DMACA), quercetin, Trolox and (+)-catechin were purchased from Fluka/Sigma-Aldrich (Madrid, Spain). Cyanidin-3-O-rutinoside was acquired from PhytoLab (Vestenbergsgreuth, Germany). 2,2′-Azobis (2-methylpropionamidine) dihydrochloride was purchased from Acros Organics (Geel, Belgium). Standard compounds were individually dissolved in acetone/Milli-Q water/acetic acid (70/29.5/0.5; v/v/v) and stored at −20 °C. All standard stock solutions were newly prepared every three months.
7
Antioxidant Assays and Phytochemical Analysis
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Folin-Ciocalteu reagent, sodium carbonate, sodium nitrate, aluminium chloride, sodium hydroxide, potassium chloride, sodium acetate, vanillin, p-dimethylaminocinnamaldehyde (DMAC), iron chloride hexahydrate, 2,4,6 tripyridyl-striazine (TPTZ), 2,2-diphenyl-1-picryl-hydrazyl (DPPH), 2,2'-azino-bis-[3-ethyl-benzothiazoline]-6-sulfonic acid (ABTS), 6-hydroxy-2,5,7,8-tetramethylchroman-2carboxylic acid (Trolox), sodium phosphate monobasic, sodium phosphate dibasic, sodium chloride, potassium persulfate, catechin and gallic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). For gallic acid, catechin, epicatechin, ellagic acid, protocatechuic acid, vanillin, chlorogenic acid, all standards were HPLC grade and were purchased from Sigma-Aldrich (St. Louis, MO, USA). HPLC-grade acetonitrile was purchased from Tedia (Fairfield, OH, USA). HPLC-grade formic acid was purchased from Sigma-Aldrich (St. Louis, MO, USA). All other reagents were analytical grade.
8
Extraction and Detection of Citrus Phytochemicals
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Citrus leaves were ground in liquid nitrogen and DNA and RNA were extracted using the DNeasy Plant Mini Kit and the RNeasy Plant Mini Kit (QIAGEN) according to the manufacturer's instructions. Fruit juice was obtained using a conventional citrus squeezer and pH was measured in three replicates with a standard combination Ag/AgCl pH electrode. The presence of anthocyanins in young leaves and flowers was determined by visual inspection and confirmed measuring the absorbance at 530 nM of acidified methanol extracts. The presence of proanthocyanidins in seeds was determined by visual inspection and confirmed by staining for 30 min with 0.3% (w/v) DMACA (p-Dimethylaminocinnamaldehyde, Sigma) in methanol and 6 M HCl (1:1, v/v) followed by several washing steps with 70% ethanol.
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