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Article ; Online: Novel dummy molecularly imprinted polymer for simultaneous solid-phase extraction of stanozolol metabolites from urine.

Farag, Yomna G / Hanafi, Rasha S / Hammam, Mennatallah A

2024

Abstract: Stanozolol, a synthetic derivative of testosterone, is one of the common doping drugs among athletes and bodybuilders. It is metabolized to a large extent and metabolites are detected in urine for a longer duration than the parent compound. In this study, ...

Abstract	Stanozolol, a synthetic derivative of testosterone, is one of the common doping drugs among athletes and bodybuilders. It is metabolized to a large extent and metabolites are detected in urine for a longer duration than the parent compound. In this study, a novel dummy molecularly imprinted polymer (DMIP) is developed as a sorbent for solid-phase extraction of stanozolol metabolites from spiked human urine samples. The optimized DMIP is composed of stanozolol as the dummy template, methacrylic acid as the functional monomer, and ethylene glycol dimethacrylate as the cross-linker in a ratio of 1:10:80. The extracted analytes were quantitively determined using a newly developed and validated ultrahigh-performance liquid chromatography tandem mass spectrometry method, where the limits of detection and quantitation were 0.91 and 1.81 ng mL
Language	English
Publishing date	2024-04-25
Publishing country	Germany
Document type	Journal Article
ZDB-ID	201093-8
ISSN	1618-2650 ; 0016-1152 ; 0372-7920
ISSN (online)	1618-2650
ISSN	0016-1152 ; 0372-7920
DOI	10.1007/s00216-024-05285-x
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Article ; Online: Quality-by-design approach for development of aqueous headspace microextraction GC-MS method for targeted metabolomics of small aldehydes in plasma of cardiovascular patients.

Hanafi, Rasha S / Lämmerhofer, Michael

Analytica chimica acta

2022 Volume 1221, Page(s) 340176

Abstract: Lipid peroxidation products, such as short chain aldehydes, are powerful biomarkers of oxidative stress, due to the advantage of long lifetime compared to other metabolites of the lipidome. This work proposes an advanced combined derivatization/solvent- ... ...

Abstract	Lipid peroxidation products, such as short chain aldehydes, are powerful biomarkers of oxidative stress, due to the advantage of long lifetime compared to other metabolites of the lipidome. This work proposes an advanced combined derivatization/solvent-less extraction procedure from plasma followed by rapid Gas Chromatography with Mass Spectrometric detection (GC-MS). A new sample pretreatment protocol is presented which is based on a combination of aldehyde derivatization with methoxyamine under fully aqueous-based conditions of diluted plasma samples followed by headspace solid-phase microextraction (HS-SPME) which is faster compared to methods in the literature serving the same purpose. Being the smallest oximation reagent, methoxyamine derivatization does not require a silylation step of hydroxyl groups as customary and made it possible to have the shortest run times for this series of aldehydes by GC-MS. A Response Surface Methodology (RSM) is employed to optimize the HS-SPME of the aldehyde methoximes to provide insights into the Design Space (DS) of HS-SPME of aldehydes of variable chain lengths and unsaturation. The workflow includes a Quality by Design (QbD) approach for optimization of sample microextraction and derivatization methodology under fully aqueous conditions, in contrast to all reported non-aqueous tedious and long extraction methods in the literature followed by development of a rapid GC-MS assay. The optimal sample preparation obtained from the RSM, and multiple linear regression procedure involved addition of 15 mg methoxyamine (CH
MeSH term(s)	Aldehydes/analysis ; Gas Chromatography-Mass Spectrometry/methods ; Humans ; Malondialdehyde ; Metabolomics ; Solid Phase Microextraction/methods
Chemical Substances	Aldehydes ; Malondialdehyde (4Y8F71G49Q)
Language	English
Publishing date	2022-07-16
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1483436-4
ISSN	1873-4324 ; 0003-2670
ISSN (online)	1873-4324
ISSN	0003-2670
DOI	10.1016/j.aca.2022.340176
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Article ; Online: Quality by design approach for enantioseparation of terbutaline and its sulfate conjugate metabolite for bioanalytical application using supercritical fluid chromatography.

Aboushady, Dina / Hanafi, Rasha S / Parr, Maria K

Journal of chromatography. A

2022 Volume 1676, Page(s) 463285

Abstract: Terbutaline is mainly metabolized by sulfoconjugation stereoselectively, favoring its (S)-(+) enantiomer. Reported chiral separations of Terbutaline enantiomers were achieved by various chromatographic methods. However, the simultaneous enantioseparation ...

Abstract	Terbutaline is mainly metabolized by sulfoconjugation stereoselectively, favoring its (S)-(+) enantiomer. Reported chiral separations of Terbutaline enantiomers were achieved by various chromatographic methods. However, the simultaneous enantioseparation of Terbutaline and the monosulfate conjugate metabolites was never reported. This study aims at shedding light on the influential factors and interactions leading to successful enantioseparation of Terbutaline and its monosulfate conjugate pairs by Supercritical Fluid Chromatography (SFC) for the first time within a Quality by Design framework using Design of Experiments. The effect of molarity of mobile phase additive, mobile phase flow rate, column temperature and back pressure were evaluated. Compared to previous reports, the response surface interestingly revealed the favorability of high temperature and high flow rate up to 2.25 ml/min for resolution of the two pairs of enantiomers on polysaccharide chiral stationary phase CHIRALPAK IC. In addition, a switch in the elution order of Terbutaline and the sulfate conjugate peak pairs was observed upon elimination of the mobile phase additive where the sulfate conjugate underwent intra-molecular ionic interactions and the change in elution order was only due to TER behavior. The multifactorial interactions would not have been detected with the common one-factor-at-a-time approach during method development, demonstrating the superiority and importance of the Analytical Quality by Design frame in enantioseparation.
MeSH term(s)	Chromatography, Supercritical Fluid/methods ; Polysaccharides/chemistry ; Stereoisomerism ; Sulfates ; Terbutaline
Chemical Substances	Polysaccharides ; Sulfates ; Terbutaline (N8ONU3L3PG)
Language	English
Publishing date	2022-06-27
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1171488-8
ISSN	1873-3778 ; 0021-9673
ISSN (online)	1873-3778
ISSN	0021-9673
DOI	10.1016/j.chroma.2022.463285
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Article: Quality by design approach for enantioseparation of terbutaline and its sulfate conjugate metabolite for bioanalytical application using supercritical fluid chromatography

Aboushady, Dina / Hanafi, Rasha S. / Parr, Maria K.

Journal of chromatography. 2022 Aug. 02, v. 1676

2022

Abstract	Terbutaline is mainly metabolized by sulfoconjugation stereoselectively, favoring its (S)-(+) enantiomer. Reported chiral separations of Terbutaline enantiomers were achieved by various chromatographic methods. However, the simultaneous enantioseparation of Terbutaline and the monosulfate conjugate metabolites was never reported. This study aims at shedding light on the influential factors and interactions leading to successful enantioseparation of Terbutaline and its monosulfate conjugate pairs by Supercritical Fluid Chromatography (SFC) for the first time within a Quality by Design framework using Design of Experiments. The effect of molarity of mobile phase additive, mobile phase flow rate, column temperature and back pressure were evaluated. Compared to previous reports, the response surface interestingly revealed the favorability of high temperature and high flow rate up to 2.25 ml/min for resolution of the two pairs of enantiomers on polysaccharide chiral stationary phase CHIRALPAK IC. In addition, a switch in the elution order of Terbutaline and the sulfate conjugate peak pairs was observed upon elimination of the mobile phase additive where the sulfate conjugate underwent intra-molecular ionic interactions and the change in elution order was only due to TER behavior. The multifactorial interactions would not have been detected with the common one-factor-at-a-time approach during method development, demonstrating the superiority and importance of the Analytical Quality by Design frame in enantioseparation.
Keywords	enantiomers ; metabolites ; polysaccharides ; stereoselectivity ; sulfates ; supercritical fluid chromatography ; temperature
Language	English
Dates of publication	2022-0802
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	218139-3
ISSN	0021-9673 ; 0378-4355 ; 0376-737X
ISSN	0021-9673 ; 0378-4355 ; 0376-737X
DOI	10.1016/j.chroma.2022.463285
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Article: Multivariate approach for optimization of galactomannan extraction from seeds of Egyptian Trigonella foenum-graecum with insights on its pharmacological activities

Ammar, Mai K. / Handoussa, Heba / Hanafi, Rasha S. / El-Shazly, Mohamed / Gad, Mohamed Z.

Natural product research. 2022 Apr. 18, v. 36, no. 8

2022

Abstract: Response surface methodology (RSM), based on the central composite design (CCD), was used for the systemic optimization of galactomannan (GAL) extraction from Trigonella foenum-graecum. GAL was reported to possess a variety of pharmacological effects and ...

Abstract	Response surface methodology (RSM), based on the central composite design (CCD), was used for the systemic optimization of galactomannan (GAL) extraction from Trigonella foenum-graecum. GAL was reported to possess a variety of pharmacological effects and is commercialized as adjuvant therapy for diabetes, obesity, and hypercholesterolemia. Seven process variables were evaluated (12 experiments in a Plackett-Burman design) to screen the significant factors affecting the extraction yield. The three most significant variables were evaluated in CCD at two levels (twenty experimental designs) to obtain the utmost percentage yield. The yield of GAL extraction was influenced by the volume of the precipitating solvent to the volume of the soaking water and reached a maximum of 10.1% at a ratio of 3.633:1. Exploring the antioxidant, cytotoxic, and anti-microbial activities of GAL revealed cytotoxic activity against LS174-T colorectal cancer cells, weak antioxidant activity, and moderate antimicrobial activity against Candida tropicalis and Micrococcus species.
Keywords	Candida tropicalis ; Micrococcus ; Trigonella foenum-graecum ; adjuvants ; antimicrobial properties ; antioxidant activity ; antioxidants ; colorectal neoplasms ; cytotoxicity ; diabetes ; galactomannans ; hypercholesterolemia ; obesity ; research ; response surface methodology ; solvents ; therapeutics
Language	English
Dates of publication	2022-0418
Size	p. 2125-2128.
Publishing place	Taylor & Francis
Document type	Article
ZDB-ID	2185747-7
ISSN	1478-6427 ; 1478-6419
ISSN (online)	1478-6427
ISSN	1478-6419
DOI	10.1080/14786419.2020.1837817
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Article: Quality-by-Design Is a Tool for Quality Assurance in the Assessment of Enantioseparation of a Model Active Pharmaceutical Ingredient.

Aboushady, Dina / Parr, Maria Kristina / Hanafi, Rasha S

Pharmaceuticals (Basel, Switzerland)

2020 Volume 13, Issue 11

Abstract: The design of experiments (DoE) is one of the quality-by-design tools valued in analytical method development, not only for cost reduction and time effectiveness, but also for enabling analytical method control and understanding via a systematic workflow, ...

Abstract	The design of experiments (DoE) is one of the quality-by-design tools valued in analytical method development, not only for cost reduction and time effectiveness, but also for enabling analytical method control and understanding via a systematic workflow, leading to analytical methods with built-in quality. This work aimed at using DoE to enhance method understanding for a developed UHPLC enantioseparation of terbutaline (TER), a model chiral drug, and to define quality assurance parameters associated with using chiral mobile phase additives (CMPA). Within a response surface methodology workflow, the effect of different factors on both chiral resolution and retention was screened and optimized using Plackett-Burman and central composite designs, respectively, followed by multivariate mathematical modeling. This study was able to delimit method robustness and elucidate enantiorecognition mechanisms involved in interactions of TER with the chiral modifiers. Among many CMPAs, successful TER enantioresolution was achieved using hydroxypropyl β-cyclodextrin (HP-β-CD) added to the mobile phase as 5.4 mM HP-β-CD in 52.25 mM ammonium acetate. Yet, limited method robustness was observed upon switching between the different tested CMPA, concluding that quality can only be assured with specific minimal pre-run conditioning time with the CMPA, namely 16-column volume (60 min at 0.1 mL/min). For enantiorecognition understanding, computational molecular modeling revealed hydrogen bonding as the main binding interaction, in addition to dipole-dipole inside the CD cavity for the
Language	English
Publishing date	2020-11-04
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2193542-7
ISSN	1424-8247
ISSN	1424-8247
DOI	10.3390/ph13110364
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Article: Point-of-care testing and optimization of sample treatment for fluorometric determination of hydrogen sulphide in plasma of cardiovascular patients.

Alyan, Ahmed K / Hanafi, Rasha S / Gad, Mohamed Z

Journal of advanced research

2019 Volume 27, Page(s) 1–10

Abstract: Introduction: ...

Abstract	Introduction:
Language	English
Publishing date	2019-11-28
Publishing country	Egypt
Document type	Journal Article ; Review
ZDB-ID	2541849-X
ISSN	2090-1224 ; 2090-1232
ISSN (online)	2090-1224
ISSN	2090-1232
DOI	10.1016/j.jare.2019.11.010
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Article: Niazimicin: A thiocarbamate glycoside from Moringa oleifera Lam. seeds with a novel neuroprotective activity

Abdelsayed, Eman M. / Medhat, Dalia / Mandour, Yasmine M. / Hanafi, Rasha S. / Motaal, Amira Abdel

Journal of food biochemistry. 2021 Dec., v. 45, no. 12

2021

Abstract: Moringa oleifera (MO) known as the miracle tree is a famous nutritional source in many countries. In this study, the neuroprotective activity of MO seeds was investigated. Fractions of the 70% ethanol seed extract of MO were injected at a dose of 250 mg ... ...

Abstract	Moringa oleifera (MO) known as the miracle tree is a famous nutritional source in many countries. In this study, the neuroprotective activity of MO seeds was investigated. Fractions of the 70% ethanol seed extract of MO were injected at a dose of 250 mg kg⁻¹ day⁻¹ to albino rats for 15 days, after‐which induction of dementia was done using 100 mg/kg AlCl₃ over 30 days. Results revealed that all fractions ameliorated the effects of AlCl₃ where methylene chloride and ethyl acetate fractions, containing the major bioactive compound niazimicin (NZ), showed the best activities. Biological investigations proved NZ to be a highly potent neuroprotective drug lead as a first report, by causing a decrease in the levels of malondialdehyde, cholinesterase, nitric oxide (NO) and amyloid β by 47%, 34%, 53% and 59%, respectively, and increasing glutathione levels by 54%. Molecular docking studies suggested NZ neuroprotective effects to be mediated by inhibition of caspase‐3 and inducible nitric oxide synthase enzymes. PRACTICAL APPLICATIONS: The current findings present the neuroprotective effect of Moringa oleifera seeds consumed as a food supplement and in daily diet. In addition, niazimicin is a promising lead for the development of novel agents against Alzheimer’s disease as seen by the reported results.
Keywords	Moringa oleifera ; albino ; amyloid ; bioactive compounds ; caspase-3 ; cholinesterase ; dietary nutrient sources ; dietary supplements ; ethanol ; ethyl acetate ; glutathione ; glycosides ; inducible nitric oxide synthase ; malondialdehyde ; methylene chloride ; neuroprotective effect ; nitric oxide ; seed extracts ; trees
Language	English
Dates of publication	2021-12
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	433846-7
ISSN	1745-4514 ; 0145-8884
ISSN (online)	1745-4514
ISSN	0145-8884
DOI	10.1111/jfbc.13992
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Article ; Online: An intronic DHCR7 genetic polymorphism associates with vitamin D serum level and incidence of acute coronary syndrome.

Elbehairy, Mariam M / Abdelnasser, Hala Y / Hanafi, Rasha S / Hassanein, Sally I / Gad, Mohamed Z

Steroids

2021 Volume 169, Page(s) 108825

Abstract: Introduction: Vitamin D deficiency has been linked to cardiovascular pathologies including acute coronary syndrome (ACS). Polymorphisms in vitamin D associated genes have been confounding to vitamin D serum levels and pathological predispositions. 7- ... ...

Abstract	Introduction: Vitamin D deficiency has been linked to cardiovascular pathologies including acute coronary syndrome (ACS). Polymorphisms in vitamin D associated genes have been confounding to vitamin D serum levels and pathological predispositions. 7-hydrocholesterol is a common precursor in cholesterol and vitamin D synthesis. DHCR7/NADSYN1 genetic locus expresses 7-hydrocholesterol reductase (DHCR7), an enzyme that recruits 7-hydrocholesterol in cholesterol biosynthesis, and NAD synthetase 1 (NADSYN1), which participates in the hydroxylation of 25 hydroxyvitamin D. Aim: This study aims to correlate two polymorphisms in the DHCR7/NADSYN1 genetic locus with levels of circulatory vitamin D and the presentation of ACS in an Egyptian population. Methods: In a case control study, 189 ACS patients and 106 healthy control subjects were genotyped for SNPs rs11606033 of the DHCR7 gene and rs2276360 of the NADSYN1 gene using the amplification-refractory mutation system (ARMS). The levels of 25(OH)D2 and 25(OH)D3 were measured using an in-house developed and validated ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) based protocol. Results: ACS patients have significantly lower levels of circulating vitamin D in comparison to healthy controls. Allele A of the DHCR7 polymorphism was found to correlate with serum vitamin D deficiency and incidence of ACS classes: NSTEMI, STEMI and unstable angina, when compared to allele G. On the other hand, the NADSYN1 polymorphism rs2276360 correlated with serum 25(OH)D3 deficiency. Yet, no significant correlation was found with incidences of ACS. Conclusion: We conclude that rs11606033, which is an intronic SNP between exon 4 and exon 5 of the DHCR7 gene, influences vitamin D serum abundance and more importantly ACS incidence.
MeSH term(s)	Acute Coronary Syndrome ; Case-Control Studies ; Humans ; Incidence ; Male ; Middle Aged
Language	English
Publishing date	2021-03-17
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80312-1
ISSN	1878-5867 ; 0039-128X
ISSN (online)	1878-5867
ISSN	0039-128X
DOI	10.1016/j.steroids.2021.108825
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Article ; Online: Niazimicin: A thiocarbamate glycoside from Moringa oleifera Lam. seeds with a novel neuroprotective activity.

Abdelsayed, Eman M / Medhat, Dalia / Mandour, Yasmine M / Hanafi, Rasha S / Motaal, Amira Abdel

Journal of food biochemistry

2021 Volume 45, Issue 12, Page(s) e13992

Abstract	Moringa oleifera (MO) known as the miracle tree is a famous nutritional source in many countries. In this study, the neuroprotective activity of MO seeds was investigated. Fractions of the 70% ethanol seed extract of MO were injected at a dose of 250 mg kg
MeSH term(s)	Amyloid beta-Peptides ; Animals ; Glycosides/pharmacology ; Molecular Docking Simulation ; Moringa oleifera ; Plant Extracts/pharmacology ; Rats ; Thiocarbamates
Chemical Substances	Amyloid beta-Peptides ; Glycosides ; Plant Extracts ; Thiocarbamates
Language	English
Publishing date	2021-11-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	433846-7
ISSN	1745-4514 ; 0145-8884
ISSN (online)	1745-4514
ISSN	0145-8884
DOI	10.1111/jfbc.13992
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