Modernization of Chinese herbal compound and the high performance liquid chromatography tandem mass spectrometry (HPLC-MS)

Chinese herbal compound is playing an important role on curing human diseases.And it has been a trend that Chinese herbal compound is being used all over the world in 21 century.However,our Chinese herbal compound is facing serious challenge for the lack of canonical system of quality criterion for Chinese herbal compound so it has been a urgent problem to set up the quality control standards and reveal therapeutic basis of Chinese herbal compound.In order to give full play to the advantages of Chinese herbal compound,modern scientific and technological is used to research of Chinese herbal compound,especially the high performance liquid chromatography tandem mass spectrometry(HPLC-MS),because it is high sensitive,rapid,and obtain more information.It is very necessary that HPLC-MS is uesed to elucidate the effective components of basic substances of Chinese Herbal Compound,and endow traditional Chinese medicine with modern scientific connotation.

Quantitative‑Profling Method of Serum Steroid Hormones by Hydroxylamine‑Derivatization HPLC-MS

A sensitive and rapid high performance liquid chromatography-mass spectrometry(HPLC-MS)method was developed and validated for simultaneous quantifcation of ten steroid hormones,including estrogens,androgens,progesterones,and corticosteroids four classes of steroids.The following ten steroid hormones were analyzed:progesterone,21-deoxycortisol,estrone,4-androstenedione,testosterone,dihydro-testosterone,androstenone,dehydroepiandrosterone,corticosterone and cortisone.Stable deuterated isotopes were used as internal standards for quantifcation.Sample preparation with and without derivatization were performed after liquid-liquid extraction,and the corresponding results were compared according to sensitivity and selectivity.Hydroxylamine derivatization was found to improve the ionization efciency of the analytes for electrospray ionization MS analysis.The gradient of mobile phase and experimental parameters for HPLC separation were optimized.The lower limits of quantifcation were in the range of 0.05-5 ng mL^(−1) with wide linear range for the ten steroid hormones.The intra-day precision<11.1%and recovery of 84.5-120% with negligible matrix efect were achieved,where within the acceptance limits of the FDA guideline.Total HPLC-MS analysis time was 6 min.This method enables simultaneous quantifcation of steroids in human serum.It will be helpful for the serum steroid profling in order to understand various endocrinology diseases.

HPLC-MS/MS Determination of Oleandrin and Adynerin in Blood with Solid Phase Supported Liquid-Liquid Extraction

[Objectives]To optimize the determination method of oleandrin and adynerin in blood. [Methods]High performance liquid chromatography-mass spectrometry( HPLC-MS/MS) was applied to determine oleandrin and adynerin in blood. The blood sample was dispersed and fixed on a solid phase supported liquid-liquid extraction column and eluted with ethyl acetate. The resulting eluent was used for chromatographic separation with Kinetex C_(18) column as the separation column and gradient elution was performed using 10 mmol/L ammonium formate solution containing 0. 1%( volume fraction) formic acid and acetonitrile as the mobile phase. In the tandem mass spectrometry analysis,the detection was carried out using the electrospray positive ion source multiple reaction monitoring mode. [Results] The mass concentration of oleandrin and adynerin showed linear relationship in the range of 2-100 μg/L. The limit of detection( 3 S/N) of the method was 0. 5 μg/L.A blank sample was used as the substrate for the spike recovery test. The recovery rate was in the range of 90. 0%-98. 0%,and the relative standard deviation( RSD) of the measured values( n = 6) was in the range of 2. 1%-7. 3%. [Conclusions]The method established in this experiment has the benefits of simple pretreatment,good recovery,high sensitivity and strong specificity,and is expected to provide an ideal method for the determination of such drugs in blood.

Mass Spectrometric Structure Elucidation of the Trivalent and Pentavalent Nitrogen Contaminants of Pholcodine in the Cough Relief Medical Form Tuxidrin

In the paper “Supercritical Fluid Chromatography-Mass Spectrometry (SFC-MS) and MALDI-TOF-MS of Heterocyclic Compounds with Trivalent and Pentavalent Nitrogen in Cough Relief Medical Forms Tuxi and Cosylan” [1], the presence of morphine and other degradation products of pholcodine in cough relief medical forms of Tuxi are discussed. Tuxiis recalled from the Norwegian market by Weifa pharmaceutical company, and hence it no longer presents problems to users and health authorities there;however, the medical form Tuxidrin, which contains a significant amount of pholcodine as the active pharmacological ingredient, is still marketed. In the present paper, Tuxidrin is analyzed to determine the presence of degradation products of pholcodine. The degradation of pholcodine to morphine has been discussed previously as a factor in the development of addiction to narcotics in young persons. The structures of the contaminants in Tuxidrin, such as oxides of pholcodine, are elucidated in the present paper. The toxicity and pharmacology of oxides of alkaloids have generally not been well studied, and very little is known about the toxicity and pharmacology of the degradation (oxidation) products of pholcodine: the N-oxide and the N, N'-dioxide of pholcodine. According to Brondz and Brondz[1], the N-oxide and possibly also the N, N'-dioxide are less toxic than the original alkaloids and possess greater pharmacological activity, and hence they may be a source of useful new semisynthetic drugs. The question of possible addiction to pholcodine oxides has not been studied, and the potential of these substances to provoke allergies is unclear. The recall of Tuxi from the Norwegian marketis mainly based on the fact that pholcodine causes significantly increased levels of IgE antibodies in sensitized patients. Tuxidrin contains pholcodine and has the same negative effect as Tuxi, namely provoking allergies or even anaphylactic shock. From this point of view, Tuxidrin has no advantage over Tuxi. These two medical forms only differ in one respect: Tuxidrin requires a prescription (prescription duty medicine), but Tuxi doesnot (prescription free medicine). This aspect is also discussed in the present paper.

液相色谱-串联质谱法测定水环境中的十氯酮

建立了分析测定水环境中十氯酮的液相色谱-串联质谱法。水样经液液萃取、净化后，采用 Eclipse plus C18柱（100 mm×2.1 mm，3.5μm）分离，乙腈和水为流动相进行梯度洗脱，在电喷雾负离子多反应监测模式下进行检测，同位素内标法定量。结果表明：采用液相色谱-质谱联用技术，证实了十氯酮在甲醇中以半缩醛的形式存在，而在丙酮/乙腈中以偕二醇的形式存在。由于十氯酮极性较强，在净化时难以洗脱，并且不耐酸，所以不能与其他有机氯农药一起分析。十氯酮在5～100μg / L 范围有良好的线性关系，相关系数 r2=0.999，检出限及定量限分别为0.70 ng / L 和2.8 ng / L；在5、40和100 ng / L 3个浓度添加水平的平均回收率为95.1％～98.9％，相对标准偏差为3.85％～4.72％。本方法具有良好的灵敏度、回收率和重现性，适用于水环境中十氯酮的测定。

高效液相色谱-串联质谱法测定水产品中甲氧苄啶的残留

建立水产品中甲氧苄啶药物残留量测定的高效液相色谱-串联质谱方法。样品选择乙腈提取，用正己烷脱脂，固相萃取柱净化，采用LC-MS/MS选择反应监测（SRM）正离子模式测定进行定性、定量分析。结果表明：甲氧苄啶药物的检出限（LOD）为0.5μg/kg，定量限（LOQ）为1.0μg/kg，检测结果的相对标准偏差为1.78%-5.08%，加标回收率达到77.1%-93.5%。该方法具有比较高的重现性和选择性，在水产品中甲氧苄啶的残留测定中具有很好的应用前景。

生物表面活性剂鼠李糖脂的纯化与表征

生物表面活性剂鼠李糖脂是微生物在一定条件下产生的次级代谢产物,其分子具有极性亲水基团和非极性亲油基团结构,通常表现出很高的表面活性和界面优先分配能力。可靠的分离提纯方法和成分鉴定手段是鼠李糖脂生产工艺成功的重要保证。实验通过好氧发酵培养铜绿假单胞菌CCTCC AB93066、酸沉降分离得到鼠李糖脂后,利用柱色谱提纯技术得到纯化的鼠李糖脂的单糖脂和二糖脂,最后采用高效液相色谱-质谱联用法进行成分鉴定。结果显示这两种鼠李糖脂均含有3种主要成分,其中单糖脂的主要成分为RhaC10C10、RhaC10C12-H2、RhaC10C12,二糖脂的主要成分为Rha2C10C10、Rha2C10C12-H2、Rha2C10C12。该研究结果表明,铜绿假单胞菌CCTCC AB93066是一种良好的鼠李糖脂产生菌;酸沉降-柱色谱技术可以用于鼠李糖脂的深度提纯,且有较好的效果;而高效液相色谱-质谱联用技术对鼠李糖脂成分鉴定具有灵敏度高和准确性好等优点,是一种较为可靠的检测方法。

Pharmacokinetics of oxiracetam and its degraded substance (HOPAA) after oral and intravenous administration in rats

The pharmacokinetics of oxiracetam and its degraded substance(4-hydroxy-2-oxo-1-pyrrolidine acetic acid,HOPAA)after oral and intravenous administration in rats were studied using an established UPLC-MS/MS method.Three groups of rats after an overnight fasted received 10 g/kg(n=6)oxiracetam suspensions orally,and 2 g/kg(n=6)normal or degraded oxiracetam injections intravenously via a caudal tail vein,respectively.Before the pharmacokinetic experiment,a simple safety evaluation testwas conducted on the degraded oxiracetam injections containing 16.16% HOPAA in mice.There was no mortality by a single intravenous dose of 2 g/kg of degraded oxiracetam injections within twoweeks,demonstrating that HOPAA was non-toxic in mice.Following intravenous administration of the normal injections,the plasma concentration-time curves of oxiracetam and HOPAA both showed a rapid elimination phase.The values of t_(1/2)were 3.1±1.5 h for oxiracetamand 0.8±0.2 h for HOPAA,andthemean residencetimes(MRT)were 1.2±0.1h and 0.8±0.1h,respectively.Oxiracetam and HOPAA after intravenous administration of the degraded oxiracetam injections presented elimination patterns similar to those observed in the normal injections.Oral pharmacokinetic results showed that the Tmax was less than 1.5 h for the two analytes,and both had a longer t_(1/2) and MRT than those of intravenous administration.Contents of HOPAA in three groupswere calculated based on AUC_(0-t) values of the two analytes.The quantitative change of HOPAA in vivo was also evaluated by comparing the plasma concentrations of HOPAA and oxiracetamat the same time for every group.Additionally,the values of absolute bioavailability of oxiracetam were about 8.0%and 7.4%calculated by the normal or degraded oxiracetam injections,whichwere far less than the value of 75%reported in literature,indicating the necessity of further study.

Association of 2-methoxyestradiol levels with the occurrence and development of endometrial cancer in humans

Objective The aim of the study was to determine the association of urinary levels of estradiol(E_(2))and 2-methoxyestradiol(2-MeOE_(2))with the occurrence and development of endometrial cancer.Methods In this case-control study,24-h urine specimens were collected from 28 postmenopausal patients with endometrial cancer and 28 postmenopausal healthy female controls.The concentration of 2-MeOE_(2) was determined using liquid chromatography-mass spectrometry with hollow fiber liquid-phase microextraction.The concentration of E_(2) was determined using an enzyme-linked immunosorbent assay.Results Estrogen levels were different between the patients with endometrial cancer and controls.The relative quantity of E_(2) in the case group was higher than that in the control group(P<0.05),whereas that of 2-MeOE_(2) was lower in the case group than that in the control group(P<0.05).The ratio of E_(2)-to-2-MeOE_(2) in the case group was significantly higher than that in the control group(P<0.05).Conclusion The results of this study indicate an imbalance of estrogen metabolites in endometrial carcinogenesis.Reduced 2-MeOE_(2) levels and elevated E_(2)-to-2-MeOE_(2) ratio may be used as potential biomarkers for the risk assessment of estrogen-induced endometrial cancer.

Analysis of Serum Metabolic Profile by Ultra-performance Liquid Chromatography-mass Spectrometry for Biomarkers Discovery： Application in a Pilot Study to Discriminate Patients with Tuberculosis

Background：Tuberculosis （TB） is a chronic wasting inflammatory disease characterized by multisystem involvement,which can cause metabolic derangements in afflicted patients.Metabolic signatures have been exploited in the study of several diseases.However,the serum that is successfully used in TB diagnosis on the basis of metabolic profiling is not by much.Methods：Orthogonal partial least-squares discriminant analysis was capable of distinguishing TB patients from both healthy subjects and patients with conditions other than TB.Therefore,TB-specific metabolic profiling was established.Clusters of potential biomarkers for differentiating TB active from non-TB diseases were identified using Mann-Whitney U-test.Multiple logistic regression analysis of metabolites was calculated to determine the suitable biomarker group that allows the efficient differentiation of patients with TB active from the control subjects.Results：From among 271 participants,12 metabolites were found to contribute to the distinction between the TB active group and the control groups.These metabolites were mainly involved in the metabolic pathways of the following three biomolecules：Fatty acids,amino acids,and lipids.The receiver operating characteristic curves of3D,7D,and 11D-phytanic acid,behenic acid,and threoninyl-γ-glutamate exhibited excellent efficiency with area under the curve （AUC） values of 0.904 （95% confidence interval [CI]：0.863-0.944）,0.93 （95% CI：0.893-0.966）,and 0.964 （95% CI：0.941-0.988）,respectively.The largest and smallest resulting AUCs were 0.964 and 0.720,indicating that these biomarkers may be involved in the disease mechanisms.The combination of lysophosphatidylcholine （18∶0）,behenic acid,threoninyl-γ-glutamate,and presqualene diphosphate was used to represent the most suitable biomarker group for the differentiation of patients with TB active from the control subjects,with an AUC value of 0.991.Conclusion：The metabolic analysis results identified new serum biomarkers that can distinguish TB from non-TB diseases.The metabolomics-based analysis provides specific insights into the biology of TB and may offer new avenues for TB diagnosis.

Simultaneous determination of 14β-lactam antibiotics in cosmetic products by liquid chromatography tandem mass spectrometry method

In this study,a simple and rapid high-performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS） method was established and validated to determine the 14β-lactam antibiotics in cosmetic products,including 1（ceftazidime）,2（cefaclor）, 3（cefdinir）,4（ampicillin）,5（cefalexin）,6（ceftezole）,7（cefotaxim）,8（cefradine）,9（cefuroxime）,10（cephazoline）,11 （cefathiamidine）,12（cefoperazone）,13（cafalotin）,14（piperacillin）.

Simultaneous determination of seven prohibited substances in cosmetic products by liquid chromatography-tandem mass spectrometry

In this paper,we developed and validated a simple,sensitive,and selective high-performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS） method to identify and measure the following prohibited substances that may be found in cosmetic products：minoxidil,hydrocortisone, spironolactone,estrone,canrenone,triamcinolone acetonide and progesterone.Chromatographic separation was performed on a Waters Symmetry C18（100 mm×2.1 mm,3.5μm particle size） with a gradient elution system composed of 0.2%（v/v） formic acid aqueous solution and methanol containing 0.2%（v/v） formic acid at a flow rate of 0.3 mL/min.The substances were detected using a triple quadrupole mass spectrometer in the multiple reaction monitoring mode with an electrospray ionization source.All of the calibration curves showed good linearity（r 〉 0.999） within the tested concentration ranges.The limit of detection was 〈25 pg.The relative standard deviations for intraday precision for each of the prohibited substances were 〈3.5%at two concentration levels（2μg/g,10μg/g）. The relative recovery rate for each of the prohibited substances ranged from 91.8%to 111%at three concentration levels（0.1μg/g,2μg/g,10μg/g）,including the limit of quantification.In conclusion,we have developed and validated a method that can identify seven prohibited substances in cosmetic products.

Photocatalytic degradation of methyl orange using ZnO/TiO_(2) composites

ZnO/TiO_(2)composites were synthesized by using the solvothermal method and ultrasonic precipitation followed by heat treatment in order to investigate their photocatalytic degradation of methyl orange(MO)in aqueous suspension under UV irradiation.The composition and surface structure of the catalyst were characterized by X-ray diffraction(XRD),field emission scanning electron microscope(FE-SEM),and transmission electron microscopy(TEM).The degradation efficiencies of MO at various pH values were obtained.The highest degradation efficiencies were obtained before 30 min and after 60 min at pH 11.0 and pH 2.0,respectively.A sample analysis was conducted using liquid chromatography coupled with electrospray ionization ion-trap mass spectrometry.Six intermediates were found during the photocatalytic degradation process of quinonoid MO.The degradation pathway of quinonoid MO was also proposed.

Evaluation of the hepatoprotective and antioxidant activities of Rubus parvifolius L.

The hepatoprotective and antioxidant activities of the n-butanol extract of Rubus parvifolius L. (RPL), a widely used medicinal plant, were evaluated. Results demonstrated that RPL extract possessed pronounced hepatoprotective effects against carbon tetrachloride (CCl4)-induced hepatic injury in mice, which was at least partially attributed to its strong antioxidant capacity. Treatment with RPL extract markedly attenuated the increases in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels caused by CCl4 intoxication. It also significantly prevented the decrease in superoxide dismutase (SOD) activity and the increase in malondialdehyde (MDA) content of liver tissue. Meanwhile, histopathological changes of hepatic damage were also remarkably ameliorated. Phytochemical analysis based on high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) revealed the presence of various phenolic compounds, including caffeic acid conjugates, ellagic acid glycosides, and flavonol glycosides, which might be responsible for the hepatoprotective and antioxidant activities of RPL.