Super Antibiotics, Part II. Hyperforin, Mass Spectroscopy (MS) and Gas Chromatography-Mass Spectrometry (GC-MS), Evidence of Permeability of the Blood-Testis Barrier (BTB) and the Blood-Brain Barrier (BBB) to Hyperforin

In the first article of this series, we presented some evidence of hyperforin as an antibiotic, antiprotozoal, antiviral, anticancer, and immunomodulatory substance. In the present article, evidence of the permeability of the blood-testis barrier (BTB) and blood-brain barrier (BBB) to hyperforin and its distribution in other organs of the domestic pig (Sus scrofa domesticus) are revealed. Seven-month-old male boars with a body mass of 100 kg were fed a diet containing hyperforin. Organs were surgically removed under anesthesia. Organs were suitable prepared and extracted, and then analyzed using gas chromatography-mass spectrometry with supersonic molecular beams (GC-MS with SMB). The presence of hyperforin was recorded in all organs and body fluids. Special attention was paid to the evaluation of the presence of hyperforin in the brain and testes of experimental animals. The presence of hyperforin in the brain and testes of experimental animals was established by GC-MS with SMB. The results are of interest because penicillin and numerous other antibiotics cannot pass through the BTB or BBB if healthy or non-inflamed, which limits their use in patients with meningitis and gonorrhea. The findings are also of interest in cases of penicillin- and multi-antibiotic-resistant bacterial infections.

区分动植物油的气相色谱-质谱分析

The discrimination between animal oil and vegetable oil was analyzed by gas chromatography-mass spectroscopy(GC-MS) using esterifying oils under the presence of KOH.Six main fatty acids(myristic acid,palmitoleic acid,palmitic acid,linoleic acid,oleinic acid and stearic acid) were determined.It was found that animal oil and vegetable oil not only has a difference in components and relative content,but also has a difference in unsaturation.According to all of these differences,animal oil can be discriminated from vegetable oil.

Study on GC-MS fingerprint of petroleum ether fraction of Shenqi Jiangtang Granules

Objective To establish gas chromatography-mass spectrometry(GC-MS)fingerprint method for the petroleum ether fraction of Shenqi Jiangtang Granules(SQJTG)and evaluate the product quality.Methods The GC-MS fingerprint of petroleum ether fraction of SQJTG was established by GC-MS,and the chemical components corresponding to the fingerprint peaks were structurally identified on NIST2014.The batch consistency of SQJTG products was evaluated based on the chemical composition of petroleum ether parts by using fingerprint similarity evaluation and Principal components analysis(PCA)technology.At the same time,Hotelling's T2 and DMODX statistics are used to set the control range for the quality of different batches of products.Results Twenty-two components were identified from the petroleum ether part of SQJTG,accounting for 60.94%of the total components separated.The similarity of fingerprints of petroleum ether parts of 24 batches of SQJTG was greater than 0.95.The PCA of 24 batches of samples were all under the control limits of Hotellin’s T2 and DMODX statistics,indicating that the petroleum ether parts of different batches of SQJTG were consistent.Conclusion The developed GC-MS fingerprint method can be used to evaluate the quality of SQJTG.

A Metabolomics Study of the Volatile Oil from Prunella vulgaris L.on Pelvic Inflammatory Disease

Objective Pelvic inflammatory disease(PID)is one of the most common gynaecological diseases.Here,this thesis aims to investigate the therapeutic effects of Prunella vulgaris L.oil on the PID by using metabolomics based on gas chromatographymass spectrometry(GC-MS)to address this challenge.Methods First,measurements of pro-inflammatory cytokines and histological analysis of the uterus were conducted to validate the successful generation of a PID rat model.Furthermore,the volatile oil from Prunella vulgaris L.was administered to treat PID rats.Serum samples were collected before and after treatment and analyzed by GC-MS to generate metabolite profiles for each sample.The information generated from the qualitative and quantitative analysis of these metabolites was applied to distinguish between the PID model and normal control groups.Results Some metabolites,such as acetic acid,succinic acid,glyceric acid,(R*,S*)-3,4-dihydroxybutanoic acid,3-hydroxyphenylacetic acid,D-ribose and myo-inositol showed a higher contribution in the classification model;thus,they can be considered as potential biomarkers.Furthermore,the therapeutic effect of the volatile oil extracted from Prunella vulgaris L.could also be visualized using GC-MS-based metabolomics.Conclusions The results show that metabolomics studies are invaluable for disease diagnosis and therapeutic effect estimation.

Effects of Fuke Qianjin Formula on hormones and their receptors and metabonomics study in uterine fibroids model rats

Objective To investigate the effects of different fractions from Fuke Qianjin Formula(妇科千金方,FKQJF)on uterine leiomyoma(UL)to determine the best fraction.Methods FKQJF was extracted and isolated to obtain polysaccharides(FKP),flavonoids(FKF),and grease(FKG).140 female SPF SD rats were divided into 14 groups[model(MOD),normal control(NC),Gouliuqing(GLQ),Mifepristone(MFST),FKQJF,low,medium,and high dose of polysaccharides(l-FKP,m-FKP,and h-FKP),low,medium,and high dose of flavonoids(l-FKF,m-FKF,and h-FKF),low,medium,and high dose of grease(l-FKG,m-FKG,and h-FKG)],and uterine fibroids model rats were treated with drugs for four weeks.Serum levels of estrogen and progesterone were measured using enzyme-linked immunoassay assay(ELISA)kits.The expression of estrogen receptor(ER-α,ER-β)and progesterone receptor(PR)in the uterus was observed using immunohistochemistry(IHC).Serum metabolite profiles and FKG were analyzed using gas chromatography-mass spectrometry(GC-MS).Results FKQJF,h-FKF,m-FKG,and h-FKG significantly downregulated the estrogen level in the uterine fibroid model rats(P<0.01).FKQJF,h-FKF,and h-FKG significantly reduced the level of progesterone in the uterine fibroid model rats(P<0.01).The levels of ER-α,ER-β,and PR in uterine fibroid model rats were significantly decreased by FKQJF and h-FKG(P<0.01).The levels of ER-α,ER-β,and PR in the fibroid model rats were decreased by m-FKG(P<0.05).Additionally,serum metabolism results revealed that h-FKG and FKQJF could regulate related endogenous metabolites and make the pathological indices of uterine fibroids in rats close to the normal group.Forty-six components were identified in the oil,accounting for 91.97%of the total oil components.Conclusion FKQJF and h-FKG showed a significant anti-myoma activity and significantly improved the pathological state of the uterus in rats with hysteromyoma.The mechanism of action may be related to the regulation of estrogen progesterone and its receptor in uterine fibroid model animals.These findings proved the effect of FKQJF on uterine leiomyoma and provided an experimental basis for its clinical research and application.

Exploring Chemovar-Specific Cannabis Extracts Quantification and Evaluation of Cytotoxic Compounds for Targeting Glioblastoma Multiforme

Glioblastoma Multiforme (GBM) represents one of the most aggressive and metastatic brain tumors, with a dismal success rate of less than three percent after five years, particularly in tumors with active immune checkpoints. This necessitates the development of targeted endogenous agents for precise GBM treatment. Previous experiments utilizing Chemovar Specific Cannabis Extractions (CSCEs), fractionated with polar solvents and quantified using Liquid and Gas Column Chromatography combined with Mass Spectrometry (LC/GCMS), have shown reduced viability and motility in human GBM cell lines. However, the complexity of the botanical substance has hindered the personalization of standard cannabis medicines for GBM due to unknown synergistic effects of multiple compounds. To address this limitation, our study focuses on exposing AM251 cells to chemovar fractions extracted using a non-polar solvent, thereby isolating a broader spectrum of constituents. By employing LC/GCMS in conjunction with Nuclear Magnetic Resonance (NMR), we have identified and quantified nine* compounds present in the non-polar CSCE that exhibit significant efficacy (0.1 μM) in inducing cytotoxicity* in GBM tumor cells. Conversely, the polar fraction in our experiment did not demonstrate efficacy against UM251 cells. The quantification of individual compounds within a cannabis extraction that selectively induces cell death in brain tumors holds promise for guiding future research and facilitating the development of a standardized CSCE for GBM therapy.

Effects of Protein on the Detection of Chlorobenzenes in Fish and Their Elimination by Modified QuEChERS Method

Molecular fluorescence spectrometry, resonance Rayleigh scattering and gas chromatography-mass spectroscopy（GC-MS） were used to study the effect of pure fish serum albumin（FSA） as the model protein on the extraction of three chlorobenzenes（CBs： 1,3-DCB, 1,2-DCB and 1,2,4-TCB; DCB=dichlorobenzene; TCB=trichlorobenzene） in fish samples. The results show that there was a strong binding effect between CB and FSA. In an aqueous solution of 90%（volume fraction） acetone, a slow but full protein denaturation might take place, which would cause the unfolding of protein and the releasing of CBs. Based on these results, a QuEChERS（quick, easy, cheap, effective, rugged and safe） method was modified by replacing the traditional acetonitrile with acetone aqueous solution in the present work. This modified QuEChERS method was applied in the determination of CBs in fish samples. The spiked recoveries and the limits of detection were 80.4%--118.3% and 2.4--7.3 ng/g, respectively. This paper proposes a new strategy by slowing down the protein denaturation and releasing bound organic compounds to enhance the extraction efficiency of CBs in fish samples.

Disubstituted piperazine analogues of trifluoromethylphenylpiperazine and methylenedioxybenzylpiperazine: analytical differentiation and serotonin receptor binding studies

A series of N,N-disubstituted piperazines were synthesized containing the structural elements of both methylenedioxybenzylpiperazine (MDBP) and trifluoromethylphenylpiperazine (TFMPP)in a single molecule.These six potential designer drug molecules having a regioisomeric relationship were compared in gas chromatography-mass spectrometry (GC-MS),gas chromatography-infrared spectroscopy and serotonin receptor affinity studies.These compounds were separated by capillary gas chromatography on an Rxi(R)-17Sil MS stationary phase film and the elution order appears to be determined by the position of aromatic ring substitution.The majority of electron ionization mass spectral fragment ions occur via processes initiated by one of the two nitrogen atoms of the piperazine ring.The major electron ionization mass spectrometry (El-MS) fragment ions observed in all six of these regioisomeric substances occur at m/z =364,229,163 and 135.The relative intensity of the various fragment ions is also equivalent in each of the six El-MS spectra.The vapour phase infrared spectra provide a number of absorption bands to differentiate among the six individual compounds on this regioisomeric set.Thus,the mass spectra place these compounds into a single group and the vapour phase infrared spectra differentiate among the six regioisomeric possibilities.All of the TFMPP-MDBP regioisomers displayed significant binding to 5-HT2B receptors and in contrast to 3-TFMPP,most of these TFMPP-MDBP isomers did not show significant binding at 5-HT1 receptor subtypes.Only the 3-TFMPP-3,4-MDBP (Compound 5) isomer displayed affinity comparable to 3-TFMPP at 5-HT1A receptors (Ki =188 nmol/L).

GC-MS and FT-IR analysis of a coastal medicinal plant-Hyptis suaveolens(L.)Poit

Objective:To investigate the bioactive components of a coastal medicinal plant,Hyptis suaveolens(L.)Poit.(H.suaveolens)leaves using fourier transform-infrared spectroscopy and gas chromatography-mass spectrometer(GC-MS).Methods:The chemical compositions of the ethanol extract of whole plant of H.suaveolens was investigated using PerkinElmer GC-MS,while the mass spectra of the compounds found in the extract was matched with the National Institute of Standard and Technology library.Results:The results of fourier transform-infrared spectroscopy analysis confirmed the presence of secondary alcohols,phenols,alkanes,alkynes,aromatics,nitro compounds and aliphatic compounds.GC-MS analysis of the ethanolic extract revealed the existence of 30 phytochemical compounds.5,5-Dimethylimidazolidin-2,4-diamine(20.35%)was found to be the major compound.Conclusions:The results of this study offer a platform to use H.suaveolens leaves as herbal alternative for various diseases.