Free PA contents were determined as described by [40 (link)]. In brief, 100 mg of whole root samples were homogenized in 5% perchloric acid. After centrifugation, 0.5 mL of the supernatant was neutralized with 0.4 mL of 2 M NaOH, then the PAs were derivatized with 10 µL of benzoyl chloride. The benzoylated polyamines were separated by HPLC system (JASCO, HPLC system, Japan) equipped with reverse-phase C18 column (Apex octadecyl, 5 µm; 4.6 mm × 250 mm), eluent was 45% (v/v) acetonitrile/water, with flow rate at 0.5 mL/min monitored with UV detector at 254 nm. Then, 20 µL injection of standards and samples were analyzed. The applied standards were Put, Spd, and Spm in the form of hydrochlorides (Sigma-Aldrich, Germany). The results are the means of three independent biological samples expressed in nmol g−1 fresh weight−1.
Hydrochlorides
Manufactured by Merck Group
Sourced in Germany, United States
Hydrochlorides are a class of chemical compounds that contain a hydrogen chloride (HCl) molecule. They are commonly used as reagents in various laboratory applications, such as pH adjustment, salt formation, and derivatization reactions. Hydrochlorides can be found in a wide range of forms, including powders, solutions, and crystalline solids.
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2 protocols using hydrochlorides
1
Polyamine Quantification in Plant Roots
2
Quantification of Polyamines in Plant Leaves
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Free PA contents were determined as described by Szepesi et al. (2009) . In brief, 200 mg of leaf samples were homogenized in 5% perchloric acid. After centrifugation, 2.5 mL of the supernatant was neutralized with 1 mL of 2 M NaOH, and the PAs were derivatized with 10 mL of benzoyl chloride. The benzoyl-PA derivates were extracted from the aqueous phase with diethyl ether. The organic phase was evaporated to dryness. The PAs were separated by highperformance liquid chromatography (HPLC) with a mixture of acetonitrile/water (45/55, v/v) (Carlo Erba, Rodano, Italy) on a reverse-phase column (Apex octadecyl, 5 mm; 250 mm  4.6 mm) at 25 C (JASCO HPLC System, Tokyo, Japan), and monitored with UV detector at 254 nm. The applied standards were Put, Spd, and Spm in the form of hydrochlorides (Sigma-Aldrich, St. Louis MO, USA).
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