An Improved High Performance Liquid Chromatographic Method for Identification and Quantization of Polyamines as Benzoylated Derivatives

Sethi, Rajat; Chava, Sai Raghuveer; Bashir, Sajid; Castro, Mauro E.

doi:10.4236/ajac.2011.24055

Cited by 15 publications

(15 citation statements)

References 25 publications

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“…The method used was the same as previously described [11]. Here, the solvent system was methanol (solvent A) and water (solvent B) with a flow rate 1.0 ml/min (method I) was throughout the isocratic method and the chromatograph for the experimental group is shown in Figure 2.…”

Section: Hplc Methodsmentioning

confidence: 99%

“…The benzoylated amines were extracted with diethyl ether, which were eluted at room temperature through a 4.6 × 250 mm, 5 µm particle size reverse-phase C 18 column which is detected at 254 nm. The time required for completing one single run was 20 minutes and is shown in An alternative method described by Schotten-Baumann benzoylation procedure (procedure III, cited in [11]) was also used as a comparison to our chromatographic procedure (Figure 4).…”

Section: Hplc Methodsmentioning

confidence: 99%

“…The polyamines were isolated from the homogenized heart tissue through centrifugation and were derivatized as previously described [11]. Benzoylated polyamines were stored at -20˚C in all these previous methods.…”

Section: Benzoylation Proceduresmentioning

confidence: 99%

See 2 more Smart Citations

Role of Polyamines in Ozone Exposed Ischemic-Reperfused Hearts

Sethi¹,

Chava²,

Bashir³

et al. 2011

AJAC

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During the Folin Ciocalteu (F-C) micro-assay for the determination of phenolics in the presence of methanol, fine solids can form. In a previous paper, we hypothesized that the interference from alcohol on the F-C reaction can be minimized depending on the particular procedure used to reach the alkalinity condition. In order to demonstrate our hypothesis we studied, by spectrophotometrically monitoring, the time-behaviour of the reactions carried out in the presence of different methanol concentrations at the same alkalinity condition from two protocols. The results showed that the interfering effect of methanol on the F-C micro-method can be affect and even prevented depending on working conditions. In particular, the formation of fine solids can be delayed, slowed down and prevented depending on the initial carbonate concentration used. We have explained why the initial carbonate concentration, used to reach the final alkalinity condition, plays an important role in the F-C reaction carried out in the presence of methanol. Moreover, the results from real-time monitoring showed that, differently from traditional F-C procedure, our procedure allows us to carry out the F-C micro method in the presence of 6% methanol, as an extreme concentration, reading the absorbance at real time 24 min. The real-time monitoring of absorbance can be considered as a useful means to explore the effect of other parameters on precipitate formation caused by the presence of methanol in the F-C reaction.

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Section: Hplc Methodsmentioning

confidence: 99%

Section: Hplc Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Role of Polyamines in Ozone Exposed Ischemic-Reperfused Hearts

Sethi¹,

Chava²,

Bashir³

et al. 2011

AJAC

Self Cite

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“…Polyamine analysis: The polyamines viz., spermidine, spermine and putrecsine was analyzed using Ultra Fast Liquid Chromatography (UFLC; Shimadzu, Japan) equipped with PDA detector 2800 (Shimadzu; Japan) and C-18 column (Sethi et al, 2011). The mobile phase based on an isocratic program which included 60% methanol as a solvents system was used where samples were run at a flow rate of 0.750 ml min -1 and visualized at 290 nm with a UV-visible detector (Fig 1).…”

Section: Seed Treatmentmentioning

confidence: 99%

Polyamine Modulation by antagonistic bacteria Bacillus subtilis in chickpea during Fusarium oxysporum f. sp. ciceri interaction

Suthar

Patel

Singh

2019

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Chickpea (Cicer arietinum L.) is pivotal source of protein for vegetarian diet, however, its productivity is adversely affected by wilt disease. Non pathogenic rhizospheric microorganism’s leads to induce resistance and are found to be effective in management of this disease. The polyamines (PAs) content and its metabolism are the key in plant microbial interaction, so the alteration in PAs viz. spermidine (SPD), spermine (SPM) and putresine (PTR) in chickpea by Bacillus substilis isolate K18 (BS-K18) effective antagonist (75%) of Fusarium oxysporum f. sp. ciceri (Foc) and having PGPR traits was analyzed under Foc stress. The higher PAs content was reported in resistant variety (WR-315) compared to susceptible variety (JG-62). The PTR was dominant PA present in chickpea, further overall root tissue reported higher PA content as compared to leaves tissue. The PA content was constitutively improved by B. subtilis seed treatment in resistant and susceptible varieties. The Foc stress leads to induction of PA content in leaves and root tissue, where its content was higher in resistant variety as compared to susceptible variety. The BS-K18 seed treatment under Foc stress leads to induction of PA content as compared to both treatments alone, the SPD and SPM were more induced in leaves and root tissue of susceptible variety whereas PTR was more induces in resistant variety. Overall, polyamines were induced up to 3 DAT then after decline suggest their early role in plant defence mechanism, further PTR was found to be dominating polyamine during chickpea-Foc interaction under BS-K18 treatment. The Bacillus subtilis seed treatment leads to improve wilt tolerance in susceptible var. JG-62 through modulation of PAs, the same mechanism also helped to enhanced effectiveness of resistant var. WR-315.

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“…The content of polyamines in the synaptoplasma was measured according to Sethi et al [29] with slight modifications. First, 350 -350 µL of the synaptoplasma was deproteinized with an equal amount of 0.6 N HC1O 4 , and the mixture was centrifuged at 15,000 × g for 20 min.…”

Section: Measurement Of Polyamines By Hplc Analysismentioning

confidence: 99%

IL-10 Gene Knockout Reduces the Expression of mGlu Receptor 1a/b and Decreases the Glutamate-Dependent Production of Nitric Oxide

Koriauli¹,

Barbakadze²,

Natsvlishvili³

et al. 2014

JBiSE

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IL-10 provides trophic and survival effects directly on neurons, promotes axonal outgrowth, and stimulates neuroregeneration. In this study, we analyzed the activities of arginase and nitric oxide synthase (NOS) in synaptoneurosomes derived from brain cortex of C57BL/6 IL-10 gene-knockout (KO) and wild-type (Wt) mice and determined that the synaptoneurosomes derived from KO mice present lower arginase II activity and lower spermine content than those derived from Wt mice, whereas the basal NOS activity in the KO synaptoneurosomes was higher than that observed in the control synaptoneurosomes. Moreover, our results indicate that the plasma membranes isolated from the KO mice brain exhibit significantly lower spermine-induced enhancement of [ 3 H] MK-801 binding than the plasma membranes from the brain of Wt mice. Glutamate increases the production of nitric oxide (NO) in Wt synaptoneurosomes in a dose-dependent manner, whereas in the KO synaptoneurosomes, this amino acid does not affect the synthesis of NO. The glutamate-dependent acceleration of NO synthesis in Wt synaptoneurosomes was abrogated by LY367385, an antagonist of mGluR1a/b. The western blot analysis of the synaptoneurosomal proteins demonstrates that the expression of the subunits of NMDAR (NMDAR2A and NMDAR2B), the level of NMDAR-bound nNOS and the expression of iNOS are not changed in KO mice and that only the level of mGluR1a/b is markedly reduced in the synaptoneurosomes of KO mice. We conclude that a neuroprotective and neuroregenerative property of IL-10, in addition to its effects on polyamine metabolism and the spermine-dependent modulation of NMDAR, may involve the regulation of mGluR1a/b expression.

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An Improved High Performance Liquid Chromatographic Method for Identification and Quantization of Polyamines as Benzoylated Derivatives

Cited by 15 publications

References 25 publications

Role of Polyamines in Ozone Exposed Ischemic-Reperfused Hearts

Role of Polyamines in Ozone Exposed Ischemic-Reperfused Hearts

Polyamine Modulation by antagonistic bacteria Bacillus subtilis in chickpea during Fusarium oxysporum f. sp. ciceri interaction

IL-10 Gene Knockout Reduces the Expression of mGlu Receptor 1a/b and Decreases the Glutamate-Dependent Production of Nitric Oxide

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