Determination of vasoactive intestinal peptide in rat brain by high-performance capillary electrophoresis

Soucheleau, Jérôme; Denoroy, Luc

doi:10.1016/0021-9673(92)87122-o

Cited by 18 publications

(8 citation statements)

References 23 publications

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“…Gradient RP-HPLC separation was performed on a Brownlee Aquapore RP300 C8 column (Applied Biosystems, Foster City, CA, USA) as previously described [22]. The mobile phase for gradient elution were composed of 0.05 O/ o TFA (phase A) and 0.0450/0 TFA-80% acetonitrile (phase B).…”

Section: Rp-hplcmentioning

confidence: 99%

“…The tissue samples were weighed, placed into 10 Volumes of 0.1 M hydrochloric acid and homogenized in a Potter-Elvehjem glass homogenizer. The homogenates were centrifuged (4000 g, 20 min, +4"C) and the supernatants were collected and submitted to solidphase extraction using disposable octadecysilyl cartridges (Sep-PakR, Waters; Milford, MA, USA), as previously described [22]. The peptide-rich eMuent was evaporated to dryness and the residue was reconstituted into 1 mL of 0.05% TFA, filtered through a 0.45 pm Durapore filter (Millipore) and injected into the HPLC system.…”

Section: Tissue Homogenization and Peptide Extractionmentioning

confidence: 99%

“…Capillary zone electrophoresis (CZE) could be such a method, since it allows the resolution of peptides according to their charge-to-mass ratio [ Despite its numerous advantages, CZE has seldom been used to determine neuropeptides in tissues or in biological fluids. We have recently reported that a commercial capillary electrophoresis instrument with UV detection can be used to detect a neuropeptide in the rat cerebral cortex [22]. In this previous work the vasoactive intestinal peptide was determined using a multi-dimensional approach including solid-phase extraction, reversedphase high-performance liquid chromatography (RP-HPLC) and finally CZE.…”

Section: Introductionmentioning

confidence: 99%

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Detection of neurotensin in tissues by capillary zone electrophoresis

Denoroy

1994

Electrophoresis

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An electrophoretic method for detecting neurotensin in tissues was developed. Homogenates of rat duodenum and adrenal glands were first extracted by solid-phase extraction and purified by reversed-phase high-performance liquid chromatography. The neurotensin-rich fraction was further analyzed by capillary zone electrophoresis (CZE) using a commercial instrument with UV detection. A minor peak was detected as neurotensin and its identity was further confirmed by performing several CZE analyses at different pH values. Such an approach could be used to analyze with good molecular specificity the neuropeptides in some human tissues.

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Section: Rp-hplcmentioning

confidence: 99%

Section: Tissue Homogenization and Peptide Extractionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Detection of neurotensin in tissues by capillary zone electrophoresis

Denoroy

1994

Electrophoresis

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“…The broad applicability of ACE is illustrated by a variety of studies on biomolecular interactions, involving peptide-drug [34], protein-polysaccharides [35][36][37], peptide-polymers [38], drug-CDs [39,40], and protein-nucleic acid [41,42]. Although several reports have been published regarding the analysis of dendrimers [43][44][45][46][47][48] or VIP [49,50] using CE, to date, no work has reported the use of ACE to study the molecular interactions between this peptide and PAMAM dendrimers.…”

Section: Introductionmentioning

confidence: 99%

Determination of binding constants of vasoactive intestinal peptide to poly(amidoamine) dendrimers designed for drug delivery using ACE

et al. 2007

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The purpose of the present paper was to study at physiological pH the affinity between vasoactive intestinal peptide (VIP) and four poly(amidoamine) dendrimers (PAMAMs) designed for drug delivery. Therefore, a fast and reproducible CE method was first developed to analyze the strongly basic peptide. To allow an accurate determination of binding constant (K) values, the ability to suppress peptide adsorption onto the silica capillary of nonpermanent coatings (poly(ethylene oxide) (PEO), low and medium relative molecular masses poly(diallyldimethylammonium chloride) (PDDA)) or poly(acrylamide) permanent coating (PAA) was evaluated. Very good intraday repeatability of VIP migration times and peak areas (0.1-0.6 and 2.9-4.9% RSD, respectively) was obtained using two of the investigated coatings (PEO and PDDA with medium molecular mass). ACE combined with these dynamic coatings was then employed to evaluate K between VIP and two amine-terminated PAMAM dendrimers of generation 2 and 5 (G2.NH2, G5.NH2) and two carboxyl-terminated PAMAM derivatives of generation 2 and 5 (G2.COOH, G5.COOH). Binding constant of (6.7 +/- 1.1) x 10(4)/M could be determined for the couple VIP/G5.NH2, while no affinity was evidenced between VIP and all other dendrimers investigated. These results suggest that G5.NH2 might be an interesting carrier for the delivery of VIP.

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“…As recently demonstrated, capillary electrophoresis (CE) is a powerful tool for the separation of biologically active materials, such as peptides, 6,7 proteins 8,9 and nucleic acids or nucleotides. 10±12 Due to the rapid separation, high resolution (several million theoretical plates) and high sensitivity, it is thought that CE might be suitable for a more detailed analysis of the very heterogeneously distributed serum lipoproteins.…”

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confidence: 99%

Capillary isotachophoretic analysis of serum lipoproteins using a carrier ampholyte as spacer ion

Inano

Tezuka²,

Miida³

et al. 2000

Ann Clin Biochem

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SUMMARY.We have developed a novel analytical method for serum lipoproteins using a commercially available capillary electrophoresis apparatus, BioFocus 3000 (Bio-Rad Laboratories Co Ltd, USA). The analytical principle is isotachophoresis (ITP), using a carrier ampholyte, BioLyte 7/9, as a spacer ion. The method allows a much higher resolution of lipoproteins than of amino acid mixtures. Serum lipoproteins are normally separated into 13±15 peaks, including some shoulder peaks. The reproducibility of repeated analysis within a day was relatively good with the coef®cient of variation within the range 0´9±1´1%. VLDL, LDL and HDL prepared by discontinuous density ultracentrifugation could be further separated by capillary ITP. This high-resolving ability of our method enabled detection of small amounts of abnormal lipoprotein species. For example, small dense LDL, which is thought to be an atherogenic lipoprotein, could be detected within the LDL group peak. Moreover, an abnormal HDL, apolipoprotein E-rich HDL, was also detected by a single analysis. These ®ndings suggest that our capillary ITP method is a useful means for detailed analysis of lipoproteins and thus for clinical diagnosis of hyperlipoproteinaemic subjects.

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Determination of vasoactive intestinal peptide in rat brain by high-performance capillary electrophoresis

Cited by 18 publications

References 23 publications

Detection of neurotensin in tissues by capillary zone electrophoresis

Detection of neurotensin in tissues by capillary zone electrophoresis

Determination of binding constants of vasoactive intestinal peptide to poly(amidoamine) dendrimers designed for drug delivery using ACE

Capillary isotachophoretic analysis of serum lipoproteins using a carrier ampholyte as spacer ion

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