Structural stability of Hib capsular polysaccharide in vaccine manufacturing / 中国生物制品学杂志

@#Objective To evaluate the stability of polyribosylribitol phosphate(PRP),the basic structure of capsular polysaccharide of Haemophilus influenzae type b(Hib),in the preparation of Hib conjugate vaccine.Methods The structures of the prepared Hib polysaccharides,polysaccharide derivatives and protein-conjugated polysaccharides were analyzed by nuclear magnetic resonance spectroscopy(NMR).Results The detection results of the prepared Hib polysaccharides,polysaccharide derivatives and protein-conjugated polysaccharides all met the requirements of relevant standards of Chinese Pharmacopoeia(VolumeⅢ,2020 edition),and the NMR spectra showed no significant change.Conclusion The basic structure PRP of the main carbohydrate antigen of Hib conjugate vaccine had no change during the vaccine manufacturing.

Progress on the new techniques and application of clinical lipid measurements / 中华检验医学杂志

New techniques in clinical lipid measurements, such as vertical auto profile, nuclear magnetic resonance spectroscopy, electrospray differential mobility analysis and liquid chromatography-mass spectrometry/mass spectrometry, are becoming increasingly mature. Clinical application of these new techniques significantly promoted the use of new lipid parameters including the particle concentrations of low-density lipoprotein/high-density lipoprotein and other lipoprotein subtype in the risk stratification of atherosclerotic cardiovascular disease and in the efficacy monitoring of lipid-regulating therapy, above progress is helpful on developing new individualized and precise lipid management strategies. This review analyzed and summarized the research progress of the new techniques for lipid measurements in recent years, aiming to provide evidence to develop new ideas for the individualized and accurate lipid management in clinical practice.

Application prospect of nuclear magnetic resonance spectroscopy in clinical laboratory examination / 中华检验医学杂志

Nuclear magnetic resonance spectroscopy (NMRS) is a branch of spectroscopy, which can be used to determine the number, type and relative position of components in the mixture. Due to its high throughput, high sensitivity and high stability, especially its "fingerprint", non-destructive and non-biased detection of metabolites, NMRS has become one of the most commonly used analytical and detection techniques in metabolomics. Based on the research of clinical laboratory application, this review briefly expounds the technical principle of nuclear magnetic resonance spectroscopy, introduces the development and latest research results of nuclear magnetic resonance spectroscopy in biomedical application fields such as blood lipid analysis, tumor detection, prediction of mental and nervous system diseases, infectious diseases, nutrition and health management, and discusses the development prospect of clinical translational medicine.

Identification of the New Psychoactive Substance Eutylone / 法医学杂志

OBJECTIVES@#To establish a method to identify unknown sample based on the combined use of Fourier transform infrared spectroscopy (FTIR), gas chromatography-quadrupole time-of-flight mass spectrometry (GC-QTOF-MS), ultra-high performance liquid chromatography-linear ion trap quadrupole-orbitrap mass spectrometry (UPLC-LTQ-Orbitrap MS) and 1H-nuclear magnetic resonance spectroscopy (1H-NMR) technique.@*METHODS@#The unknown sample was directly analyzed by FTIR. The unknown sample was dissolved in methanol solution containing internal standard SKF525A and the supernatant was detected by GC-QTOF-MS and UPLC-LTQ-Orbitrap MS. The unknown sample was dissolved in methanol-d4 solution for structural analysis of 1H-NMR.@*RESULTS@#The characteristic absorption peaks of FTIR spectra obtained from unknown sample were 1 682 (C=O bond), 1 503, 1 488, 1 436, 1 363, 1 256, 1 092, 1 035, 935, 840 and 800 cm-1, the characteristic fragment ions (m/z) of GC-QTOF-MS were 86.096 4 (base peak), 58.065 1, 149.023 5, 121.028 6 and 65.038 6, the accurate mass [M+H]+ detected by UPLC-LTQ-Orbitrap MS was 236.127 7. The sample was identified as synthetic cathinone new psychoactive substance Eutylone by 1H-NMR.@*CONCLUSIONS@#The method established in this study can be used for structural confirmation of Eutylone.

Application progress of metabolomic analysis based on nuclear magnetic resonance spectroscopy in tumor diagnosis and treatment / 中华检验医学杂志

Nuclear magnetic resonance spectroscopy is one of the main analytical techniques for detecting metabolomics, which has the advantages of simple operation, rapid detection and non-invasive feature. By monitoring the changes of metabolites in the body, it is helpful to deeply understand the mechanism of disease and play a role in the diagnosis and treatment of diseases, but its clinical application has not yet been popularized. In recent years, the application of metabolomics in tumors has increasingly become a research hotspot. Therefore, in order to provide a reference for the research and clinical application of tumor metabolomics, the nuclear magnetic resonance spectroscopy and tumor metabolomics were introduced in this paper, and the application progress of metabolomics analysis based on this technique in early tumor screening, clinical diagnosis and prognosis evaluation were reviewed in this paper.

Identification of Synthetic Cannabinoid New Psychoactive Substances 4F-MDMB-BUTINACA and MDMB-4en-PINACA / 法医学杂志

Objective To establish a method that combines a series of techniques including Fourier transform infrared spectrum （FTIR）, gas chromatography-mass spectrometry （GC-MS）, high resolution mass spectrometry and nuclear magnetic resonance spectroscopy （NMR） for identification of unknown substances. Methods The unknown samples （off-white powder and yellow crystal） seized in the actual cases were detected by FTIR, GC-MS （methanol as solvent）, high resolution mass spectrometry （methanol as solvent） and NMR （deuterated methanol as solvent）. Results The mass spectrum characteristic ions m/z of the main components in the samples measured by GC-MS were 219 （base peak）, 363, 307, 304, 275, 145, 131 and 213 （base peak）, 357, 301, 298, 269, 185, 171, 145 and 131, respectively. The accurate mass numbers [M+H]+ measured by high resolution mass spectrometry were 364.203 61 and 358.212 34, respectively. The unknown samples were identified as synthetic cannabinoid new psychoactive substances 4F-MDMB-BUTINACA and MDMB-4en-PINACA after data consultation and database retrieval and comparison, combined with infrared analysis and mass spectrometry data analysis, and their structures were confirmed by 1H-NMR. Conclusion The established multi-technology joint identification method can be used to identify 4F-MDMB-BUTINACA and MDMB-4en-PINACA in unknown samples. This method is fast, convenient, accurate, reliable and practical, and can provide reference for the identification of cases involving such substances in the future.

Detection of urine metabolites in patients with inherited metabolic diseases by nuclear magnetic resonance spectroscopy / 中华检验医学杂志

Objective:Nuclear magnetic resonance spectroscopy (NMR) was used to detect the species and content of metabolites in urine of patients with inherited metabolic diseases, and to explore the application value of NMR technology in the diagnosis of inherited metabolic diseases.Methods:Urine samples were collected from 20 patients with inherited metabolic diseases diagnosed in Xinhua Hospital, Shanghai Jiaotong University School of Medicine from March to June 2019, including 9 cases of methylmalonic acidemia (MMA). NMR pulse length-based concentration determination and Gas chromatography mass spectrometry (GC/MS) semi-quantitative method were used to detect the composition of metabolites in urine samples of patients with inherited metabolic diseases, and the levels of abnormal metabolites in the two methods were analyzed.Results:NMR technology can detect the levels of characteristic metabolites significantly increased in the urine of patients with MMA, isovalerinemia, glutaric acidemia, propionic acidemia, 3-methylcrotonyl-CoA carboxylase deficiency, ornithine carbamyltransferase deficiency, Citrin deficiency, Canavan disease, tyrosinemia and lysinuria protein intolerance. The average is 8 times of the upper limit of the reference value, and the highest is 545 times. Compared to GC/MS, NMR technology can detect the levels of various metabolites such as organic acids, amino acids and sugars. In 9 cases of untreated MMA,the median levels of methylmalonic acid and 3-hydroxypropionic acid in NMR [1 800 (180-12 000) and 50 (0-270) mmol/mol Cr] were higher than the reference values (0-31, 0-35). The median levels of methylmalonic acid and methylmalonic acid in GC/MS [136.56 (43.79-518.67) and 4.87 (1.52-7.52)] were higher than the reference values (0-4 and 0-0.7).Conclusions:NMR and GC/MS technologies are specific for the diagnosis of organic acidemia. The primary component detected by GC/MS is organic acid. NMR technology can break through this limitation and measure the level of various metabolites in urine, which provides a more theoretical basis for the diagnosis and research of inherited metabolic disease.

Establishment of a cocrystal characterization method for sacubitril valsartan sodium / 药学学报

Sacubitril valsartan sodium (LCZ696) is an ionic cocrystal drug. The purpose of this study was to explore the cocrystal features of LC696 by establishing a variety of characterization methods, and thus provide basic research data for effective quality control. The cocrystal characteristics of LCZ696 and its tablets were identified by applying analytical means including powder X-ray diffraction (PXRD), fourier transform infrared spectroscopy (FTIR), Raman spectra (RM), differential scanning calorimetry (DSC) and solid-state nuclear magnetic resonance spectroscopy (ssNMR). The crystalline water and hygroscopicity of LCZ696 were analyzed by thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), hygroscopicity test and Karl Fischer reaction method. The results show that PXRD, FTIR, DSC and ssNMR can effectively distinguish the features of LCZ696 cocrystal, sacubitril monomer, valsartan monomer, and sacubitril-valsartan (1∶1) mixture. RM can be used as a supplementary approach. Combined with the analysis by TGA, DVS, hygroscopicity test and Karl Fischer reaction method results, LCZ696 contains 2.5 crystalline water molecules and is very hygroscopic; we recommend that LCZ696 be stored in an environment with a relative humidity below 60%. By characterizing the crystal features we can establish quality control measure and evaluate the stability of the drug tablets. This study provides data in support for the establishment of the LCZ696 quality standard.

Zika virus proteins at an atomic scale: how does structural biology help us to understand and develop vaccines and drugs against Zika virus infection?

In Brazil and in other tropical areas Zika virus infection was directly associated with clinical complications as microcephaly in newborn children whose mothers were infected during pregnancy and the Guillain-Barré syndrome in adults. Recently, research has been focused on developing new vaccines and drug candidates against Zika virus infection since none of those are available. In order to contribute to vaccine and drug development efforts, it becomes important the understanding of the molecular basis of the Zika virus recognition, infection and blockade. To this purpose, it is essential the structural determination of the Zika virus proteins. The genome sequencing of the Zika virus identified ten proteins, being three structural (protein E, protein C and protein prM) and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5). Together, these proteins are the main targets for drugs and antibody recognition. Here we examine new discoveries on high-resolution structural biology of Zika virus, observing the interactions and functions of its proteins identified via state-of-art structural methodologies as X-ray crystallography, nuclear magnetic resonance spectroscopy and cryogenic electronic microscopy. The aim of the present study is to contribute to the understanding of the structural basis of Zika virus infection at an atomic level and to point out similarities and differences to others flaviviruses.(AU)

Identification of the New Psychoactive Substance Dibutylone / 法医学杂志

Objective To establish a method to identify unknown samples based on combined use of gas chromatography-mass spectrometry （GC-MS）, high resolution mass spectrometry （HRMS） and nuclear magnetic resonance spectrum （NMR） technique. Methods The unknown samples were dissolved in methanol solution containing internal standard SKF525A and detected by GC-MS and HRMS. The mixed samples were separated and purified by silica gel column chromatography, and then dissolved in methanol-d4 solution for structural analysis of 1H nuclear magnetic resonance spectroscopy （1H NMR）. Results The characteristic fragment ions （m/z） were 86.1 （base peak）, 71.2, 121.1, and 149.0, and the accurate mass number of molecular ion peak was measured by HRMS to be 236.128 89. By combined use of data analysis and database comparison, a new psychoactive substance of the cathinone class, Dibutylone, was detected in the sample, and the sample also contained a small amount of caffeine. The sample was purified, then identified using 1H NMR, and was further confirmed to be Dibutylone. In addition, the GC-MS retention time and characteristic fragment ions of the main components of the sample were consistent with those of Dibutylone reference material. Conclusion The method established in this study can be used for the identification of Dibutylone in mixed samples.

Metabolomics Changes of Serum in Rats Dying from Untypical Electric Injury / 法医学杂志

Objective To study the differential metabolites of serum in rats dying from untypical electric injury by 1H nuclear magnetic resonance （1 NMR）-based metabolomics methods, in order to provide clues for identification of death from antemortem untypical electric injury and instant postmortem electric injury. Methods Models of rats dying from untypical electric injury, instant postmortem electric injury, mechanical asphyxia, mechanical injury, and high temperature injury were established. The rats in control group were executed without any treatment. The serums of rats from every group were detected by 1H NMR-based metabolomics technology to screen differential metabolites. Results The rats dying from untypical electric injury group was compared with those from mechanical asphyxia group, mechanical injury group, high temperature injury group, and control group, respectively. Four chemical shift points with diagnostic value, and their corresponding metabolites were screened. These chemical shift points contained many small molecules, such as alcohols, phenols, sugars, amino acids, etc. The death from untypical electric injury group was compared with those from instant postmortem electric injury group and control group, and then eight chemical shift points with diagnostic value and their corresponding metabolites were screened. These chemical shift points contained small molecules, such as sugars, amino acids, esters, nucleic acids, etc. Conclusion The 1H NMR-based metabolomics technology can identify differential metabolites of serum in rats dying from untypical electric injury, therefore it may provide a basis for the diagnosis of death from untypical electric injury and the identification of antemortem electric injury and instant postmortem electric injury.

Structure of the Conjugated Linoleic Acid-Paclitaxel Conjugate Self-Assembled Nanoparticles / 中国药学杂志

OBJECTIVE: To prepare, the conjugated linoleic acid-paclitaxel conjugate self-assembled nanoparticles (CLA-PTX NPs) by nanoprecipitation. METHODS: The Dynamic light scattering, nuclear magnetic resonance spectroscopy, raman spectroscopy, fourier transform infrared spectroscopy and nitrogen element distribution of CLA-PTX NPs were studied. RESULTS: The hydroxyl groups (C-4 and C-10 of PTX) and the acetyl groups (C-1 and C-7 of PTX) were on the surface of CLA-PTX NPs, CLA carbon chain, the benzene ring (C-2 and C-3' of PTX) and the amide bond (C-3' of PTX) were inside the CLA-PTX NPs. CONCLUSION: It is speculated that the self-assembly of CLA-PTX is that the non-polar CLA carbon chain spontaneously aggregates inward due to hydrophobic interaction, and the hydrophilic oxygen-containing groups of PTX (hydroxyl group and carbonyl group) are on the surface of the nanoparticle to form nanoparticles.

Applications of nuclear magnetic resonance spectroscopy in the screening of new psychoactive substances / 中国药科大学学报

@#GC-MS and LC-MS are the main techniques used for the structural identification of new psychoactive substances at present. However, they are hard to give accurate structure information because of the hardly available corresponding reference standards and the quickly changing status of these compounds. This leads tremendous obstacle on the rapid identification of new psychoactive substances. Nuclear magnetic resonance spectroscopy is one of the most effective methods for structures identification. Therefore, NMR is especially suitable for the analysis and identification of new psychoactive substances even with rapid structural changes. This article summarizes the NMR applications for the structural analysis of new psychoactive substances including synthetic cannabinoids, synthetic cathinones, piperazines, phenethylamines, ketamine & phencyclidine-type substances, and fentanyls. It is found that the NMR signals of the main frame structure of each kind of the new psychoactive substances are basically the same. Hence, these frame structure NMR signals can provide scientific evidence for the rapid identification of new psychoactive substances. This article also look ahead the prospect for the application of LC-NMR and DOSY in new psychoactive substances, which provides new ideas for the screening of new psychoactive substances.

Link between sortase A function and cariogenicity of Streptococcus mutans: a preliminary metabolomics analysis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study intends to explore the mechanism underlying the support of sortase A (SrtA) of the cariogenicity of Streptococcus mutans (S. mutans).</p><p><b>METHODS</b>We performed a metabonomics study based on ¹H nuclear magnetic resonance spectroscopy (NMR), in which we compared the extracellular metabolites of wild-type S. mutans UA159 with those of its SrtA-deficient strain. Metabolite differences among strains were identified using a combination of principal component analysis and orthogonality partial least square discriminant analysis.</p><p><b>RESULTS</b>Several differences corresponding mostly to unknown metabolites were identified. Some amino acids such as leucine and valine (δ 0.92×10⁻⁶-1.20×10⁻⁶), lactic acid ( δ1.28×10⁻⁶), oxoglutaric acid (δ 3.00×10⁻⁶), and glycine (δ 3.60×10⁻⁶) differed among strains.</p><p><b>CONCLUSIONS</b>This work establishes the feasibility of using ¹H NMR-based metabonomics to provide leads for research into molecular factors that promote caries. The database of microbial metabolites should be also improved in further studies.</p>

Study on the Hepatoprotective Effects of Hugan Tablets Based on Serum and Liver Metabonomics / 中国药房

OBJECTIVE:To elucidate the efficacy and mechanism of Hugan tablets in hepatoprotective effects from perspective of metabolic pathways. METHODS:36 male rats were randomly divided into normal group (0.5%sodium carboxymethyl cellu-lose),model group(0.5%sodium carboxymethyl cellulose)and Hugan tablets group(1.7 g/kg),12 in each group,intragastrically administrated once a day,for 9 d. After 1 h of last administration,rats in model group and Hugan tablets group were intraperitone-ally injected 50%CCl4 peanut oil solution 1 mL/kg to induce liver injury. After 24 h of modeling,malondialdehyde(MDA),super-oxide dismutase(SOD),glutathione peroxidase(GSH-Px)levels in liver tissue of rats were detected. Nuclear magnetic resonance spectroscopy(1H-NMR)metabolomics technique was adopted to establish the serum and liver metabolite profiles of rats,and the ef-fects of Hugan tablets on changes of metabolic profile and potential biomarkers in serum and liver of rats with CCl4-induced acute liver injury were analyzed. RESULTS:Compared with normal group,MDA level in liver tissue of rats in model group was signifi-cantly increased(P<0.05),SOD and GSH-Px levels were significantly reduced(P<0.05). Both body physiology and material me-tabolism of rats were obviously changed,and levels of 11 metabolic potential biomarkers in serum and 14 metabolic potential bio-markers in liver were significantly increased/decreased (P<0.05). Compared with model group,MDA level in liver tissue in Hugan tablets group was significantly reduced(P<0.05),SOD and GSH-Px levels were significantly increased(P<0.05). Serum and liver metabolism tended to be normal,6 metabolic potential biomarkers(isoleucine,leucine,3-hydroxybutyrate,acetone,ace-toacetate,choline) in serum and 8 metabolic potential biomarkers (3-hydroxybutyrate,alanine,glutamate,pyruvate,succinate, choline,lactate,glucose)in liver got significant callback(P<0.05). CONCLUSIONS:The hepatoprotective mechanism of Hugan tablets may be associated with antioxidative stress and regula-tion of lipid metabolism,glucose metabolism and amino acid metabolism.

Historical story on natural medicine chemistry: Application of UV, IR, MS, and NMR spectra in structure elucidation of natural products / 中草药

The study on natural product chemistry plays an important role in drug development, and the structure elucidation is one of the vital tasks in the natural product chemistry research. This paper summarized the application of UV, IR, MS, and NMR spectra in the structure elucidation of natural products with 123 papers cited. This article is one of the series of historical stories on natural product chemistry published in this journal, which are reviewed and summerized. The developing future is looked forward.

Distinguishing Tendon and Ligament Fibroblasts Based on ¹H Nuclear Magnetic Resonance Spectroscopy

Tendon and ligament (T/L) have been known to be obviously different from each other in tissue level. However, due to the overlapping gene markers, distinction in cellular level has not been clearly verified yet. Recently, the use of nuclear magnetic resonance (NMR) spectroscopy has shown the potential to detect biological markers in cellular level. Therefore, in this study we applied a non-invasive technique based on NMR spectroscopy to establish biomarkers to distinguish between T/L fibroblasts. In addition the cellular morphologies and gene expression patterns were also investigated for comparison through optical microscopy and real-time polymerase chain reaction (PCR). No difference was observed from morphology and real-time PCR results, either as expected. However, we found clear differences in their metabolomic spectra using ¹H NMR spectroscopy. The calculated integral values of fatty acids (with chemical shifts at ~0.9, 1.26, 1.59, 2.05, 2.25, and 2.81 ppm), lactate (~1.33 ppm), and leucine (~2.72 ppm) were significantly different between the two types of fibroblasts. To be specific tendon group exhibited higher level of the metabolite than ligament group. In conclusion, in-cell metabolomic evaluation by NMR technique used in this study is believed to provide a promising tool in distinguishing cell types, especially T/L cells, which cannot be classified by conventional biological assays.

Metabolic Profile of Dyskinetic Cerebral Palsy Based on Metabonomics / 中国康复理论与实践

Objective To analyze the metabolic profile of children with dyskinetic cerebral palsy by metabolomics, and its abnormal metabolic pathway. Methods The serum of 10 children with dyskinetic cerebral palsy (patient group) and 7 healthy children (control group) aged 6 to 12 years were collected at clinic from May to August, 2014. The serum samples were tested by the nuclear magnetic resonance spectrometer and the spectroscopies were discriminated by partial least squares-discriminant analysis. According to the human metabolome database, the final metabolites disturbed would be figured out. Results 15 chemical shifts were defined, and 6 of them, including 2.04 ppm, 2.12 ppm, 3.00 ppm, 3.24 ppm, 3.76 ppm, 6.50 ppm, were significantly different between 2 groups (P<0.05). The KEGG Pathway Database showed that the levels of taurine, fumarate, oxaloacete, pyruvate, citrate, aspartate, succinate, malate, cysteine decreased, and the levels of glutamate, 2-oxoglutarate, glutamine, leucine, alanine increased. The abnormal metabolism was found in taurine metabolism, glutamine me-tabolism and energy metabolism pathways. Conclusion Based on metabolomics, the metabolic profile of children with dyskinetic cerebral palsy was discriminated out successfully. The further research can focus on the small molecules found out.

The grading diagnosis of the magnetic resonance imaging and magnetic resonance spectroscopy and gross motor rating scale for children with cerebral palsy / 国际儿科学杂志

Objective To investigate the grading diagnostic value of magnetic resonance spectroscopy and magnetic resonance imaging and gross motor rating scale for children with cerebral palsy.Methods Forty cases of healthy children under 1.0 ～ 2.0 years old and 42 cases of cerebral palsy children under 1.1 ～ 2.0 years old were enrolled in the study,and we compared the inspection reports on MRI and MRS statistical analysis.According to the results of MRI for dividing cerebral palsy,and MRS indexing of NAA/Cr,CHO/Cr,LAC/Cr levels were compared;different types of cerebral palsy by GMFM scores were examined.Results Nuclear magnetic resonance(MRI) and MRS could effectively distinguish typical pathological changes in the brain of children with cerebral palsy,the cerebral NAA,Cr,Cho could appear significant formant when the children detected by MRS,could be quantitatively detected with MRS.The levels of NAA/Cr、CHO/Cr、LAC/Cr of different MRS and MRI degree was different,and the difference of moderate CP and severe CP was significant (P ＜ 0.05).Compared with MRS,the accordant rates of the MRI was not ideal,But gross motor rating scale score suggested that there were differences between the cerebral palsy of the MRI and MRS degree (P ＜ 0.05).Conclusion The magnetic resonance imaging and magnetic resonance spectroscopy and gross motor rating scale can effectively improve the dividing diagnosis of cerebral palsy,and this method for different types of cerebral palsy in children has some guiding value.

The application perspective of nuclear magnetic resonance spectroscopy in laboratory medicine / 中华检验医学杂志

Molecular biomarkers could change associated with disease processes.So,the detection of metabolic markers becomes the key to early diagnosis,treatment and prognosis evaluation disease.But the detection of abundant metabolites in body becomes a crux of laboratory medicine at the same time.The recent advances in nuclear magnetic resonance (NMR) spectroscopy reveal the unequivocal value of NMR in metabolomics platforms.NMR spectroscopy has inherently distinct capabilities to identify and quantitatively measure a large amount of low concentration metabolites in a non-destructive manner.The implementation of NMR technology into a multidisciplinary approach to biomarker identification will improve the auxiliary diagnostic ability of laboratory medicine for the disease.