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1.
Article in Chinese | WPRIM | ID: wpr-920653

ABSTRACT

@#A rapid analytical method for the determination of dezocine and pethidine in hair samples using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established.After cleaned hair was extracted by grinding with methanol and ultrasonic, the final solution was analyzed by UPLC-MS/MS.The targets were gradient eluted on a Waters Acquity BEH C18 (2.1 mm × 100 mm, 1.7 μm) column with 0.1% formic acid-water and methanol as mobile phase at a flow rate of 0.4 mL/min.The ESI+ ion source and multiple reaction monitoring (MRM) were used to select the qualitative and quantitative ion pairs of dezocine and pethidine.Dezocine and pethidine showed good linearity in the range of 0.01-8 ng/mg, with the limit of detection of 0.005 ng/mg and the LOQs of 0.01 ng/mg.The accuracy, precision, matrix effect, extraction recovery, and stability all met the requirements.The established method is simple, rapid, and accurate for the qualitative and quantitative determination of dezocine and pethidine in hair, which can be applied in the case analysis of dezocine and/or pethidine abuse.

2.
China Pharmacy ; (12): 1325-1329, 2021.
Article in Chinese | WPRIM | ID: wpr-877253

ABSTRACT

OBJECTIVE:To establish the metho d for the concentration det ermination of foretinib derivative LWK- 126 in liver microsomes,and to study its metabolism stability in liver microsomes of rats ,Beagle dogs and human. METHODS :In the in vitro incubation system of liver microsomes ,LWK-126 was dissolved in liver microsomal incubation systems of rats ,Beagle dog and human initiated by reduced nicotinamide adenine dinucleotide phosphate solution. After incubation in water at 37 ℃ for 0,5,10,20, 30 and 60 min,the reaction was terminated with acetonitrile containing internal standard (1 μg/mL tolbutamide). UPLC-MS/MS method was applied to determine the concentration of LWK- 126 in the incubation systems. The determination was performed on Waters BEH C 18 column with mobile phase consisted of water (containing 0.1% formic acid )-acetonitrile(containing 0.1% formic acid)by gradient elution at the flow rate of 0.4 mL/min. The column temperature was 30 ℃,and the sample size was 2 μL. The mass spectral analysis was performed in a positive electrospray ionization mode ,and the full MS experiment was run with the selective reaction monitoring mode with a scanning range of m/z 50→1 200. Taking the concentration of LWK- 126 at 0 min as reference,the remaining percentage and the enzyme kinetic parameters were calculated. RESULTS :The linear range of LWK- 126 was 0.05-15 μg/mL(R 2=0.999 2);the lower limit of quantification was 0.05 μg/mL,and the lowest detection limit was 0.01 μg/mL. The precision,accuracy,extraction recovery and matrix effect all met the analysis requirements of biological samples. The remaining percentage of LWK- 126 in liver microsomes of human ,rats and Beagle dogs for 60 min were (33.17±4.52)%,(3.14± 6.73)%,(1.38±5.85)%;t1/2 of them were 33.15,11.76,5.62 min;the clearance rates were 38.45,118.81,245.76 μL(/ min·mg), respectively. CONCLUSIONS :The method for the content ; determination of LWK- 126 in liver microsomes is established successfully. The order of metabolic stability of LWK- 126 in 〔2016〕4015) liver microsomes of different species is human >rats>Beagle dogs.

3.
Article in Chinese | WPRIM | ID: wpr-875679

ABSTRACT

Objective To establish an UPLC-MS/MS method of voriconazole assay in human plasma for clinical therapeutic drug monitoring. Methods The plasma samples were treated with methanol to precipitate protein and the supernatants were assayed by UPLC-MS/MS. The chromatographic column was InertSustain C18 HP (3.0 mm×100 mm, 3 μm) with the mobile phase of water and acetonitrile solution (15:85) at 35 ℃ and 0.3 ml/min flow rate. ESI was used for Mass Spectrum in positive ion MRM mode with target ions m/z: 350.9/282.2(voriconazole) and m/z: 307.0/238.0(fluconazole). Results The linear range of voriconazole was 0.1–20 μg/ml (r=0.999 5). The lower limit of quantitation was 0.1 μg/ml. The extraction recovery was higher than 90%. RSDs of inter-day and intra-day were all lower than 10%. The plasma concentrations measured by this method were in the range of 0.97 to 18.7 μg/ml from 10 patients with hepatic insufficiency who were treated with voriconazole. Conclusion The established method was fast, accurate and sensitive。It can be applied for the therapeutic drug monitoring of voriconazole,and provided a good base for rationalized medication treatment in patients with hepatic insufficiency.

4.
China Pharmacy ; (12): 480-484, 2021.
Article in Chinese | WPRIM | ID: wpr-873490

ABSTRACT

OBJECTIVE:To establish the method for simultaneous determination of 5-fluorouracil(5-FU)and its metabolites 5-fluoro-5,6-dihydrouracil (5-FUH2) in human plasma ,and apply it in the clinic. METHODS :After plasma samples were processed twice by ethyl acetate ,UPLC-MS/MS method was adopted using 5-bromouracil (5-Bru) as internal standard. The determination was performed on Acquity UPLC HSS T 3 column with mobile phase consisted of methanol-water (30 ∶ 70,V/V)at the flow rate of 0.3 mL/min. The column temperature was 20 ℃,and sample size was 5 μL. An electrospray ion source was used to carry out negative ion scanning with multiple reaction monitoring. The capillary voltage was 1.5 kV;the taper hole voltage was 20 V;the desolvent temperature was 450 ℃;the desolvent air flow was 850 L/h;the cone hole gas velocity was 50 L/h. The ion transitions for quantitative analysis were m/z 129.00→41.90(5-FU),m/z 130.87→82.92(5-FUH2),m/z 189.00→42.10(5-Bru), respectively. From Aug. to Oct. 2020,10 patients with advanced colorectal cancer were treated with continuous intravenous drip of 5-FU for 46 hours were collected from Harbin Medicinal University Cancer Hospital. Steady-state plasma concentration of 5-FU and plasma concentration of 5-FUH2 were determined at 18-30 h of continuous intravenous drip. The area under the curve (AUC)for 5-FU and concentration ratio of 5-FUH2/5-FU were calculated. RESULTS :The linear range of 5-FU and 5-FUH2 were 20 to 1 000 μg/mL(R 2>0.990). The quantification limits were 20 ng/mL. RSDs of precision test were all lower than 20%,and relative error ranged ±10%. The extraction recovery and matrix effects didn ’t affect the determination of substance to be measured. Among 10 patients with advanced colorectal cancer ,the steady-state concentration of 5-FU were 180.04-622.83 ng/mL,and AUC of 5-FU ranged from 8.28 to 28.65 mg·h/L. The concentration of 5-FUH2 ranged 336.48-948.43 ng/mL,and concentration ratio of 5-FUH2/ 5-FU ranged 0.93-4.21. AUC of 5-FU in 10 patients had about 3-4 fold individual differences. CONCLUSIONS :The established method has good precision and accuracy ,high sensitivity ,and simple operation. It can be used for plasma monitoring of 5-FU in patients with advanced colorectal cancer.

5.
Article in Chinese | WPRIM | ID: wpr-888060

ABSTRACT

As anti-aging ingredients, β-nicotinamide mononucleotide(NMN) and nicotinamide adenine dinucleotide(NAD~+) have attracted worldwide attention in recent years. After oral administration, NMN can be converted into NAD~+ in vivo and the latter is the actual ingredient which exerts anti-aging effect. In order to explore the "rejuvenating and anti-aging" effect of Dendrobium officinale, which was firstly recorded in Shennong's Herbal Classic of Materia Medica, this study established the quantitative method of UPLC-MS/MS for simultaneous determination of NMN and NAD~+ in D. officinale and the congeneric species for the first time, and 34 batches of samples were detected. UPLC conditions are as follows: ACQUITY UPLC HSS T3 column(2.1 mm × 100 mm, 1.8 μm), gradient elution with acetonitrile-0.1% formic acid in water at the flow rate of 0.3 mL·min~(-1), and column temperature of 40 ℃. MS conditions were scanned electrospray ionization source and multiple reaction monitoring mode. The method was verified by systematic methodology. The mean recoveries of NMN and NAD~+ were 77.58% and 80.70%, respectively, with RSD of 3.6% and 4.3%, separately. All results showed that the content of NMN was higher in D. officinale than in the other congeneric species. Particularly, the content in fresh D. officinale stems was as high as 0.931 9 μg·g~(-1). NAD~+ was only found in D. officinale and the content was three times higher than that of NMN. This may be the reason that D. officinale topped the "nine famous anti-aging herbs". In addition, processing method influences the content of NMN and NAD~+ in Dendrobium. Specifically, the content of NMN and NAD~+ was in the order of fresh Dendrobium stems > dried Dendrobium stem segments > spiral or spring-like dried Dendrobium stems.


Subject(s)
Chromatography, High Pressure Liquid , Chromatography, Liquid , Dendrobium , NAD , Nicotinamide Mononucleotide , Tandem Mass Spectrometry
6.
Article in Chinese | WPRIM | ID: wpr-888054

ABSTRACT

The effect of intestinal flora changes on the pharmacokinetics of astragaloside Ⅳ in rats with type 2 diabetes mellitus was explored in this study. The rat model in preliminary experiment was established by high-sugar and high-fat diet combined with the intraperitoneal injection of low-dose streptozotocin(STZ). Rats were divided into model group, astragaloside Ⅳ group, berberine group and combination group(five rats in each group). After two weeks of gavage, the rats' feces was taken for 16 S rRNA sequencing of intestinal flora. Pharmacokinetic experiments were performed on astragaloside Ⅳ in the four groups one day after the preliminary experiment. Plasma samples were precipitated in methanol with ginsenoside Rb_1 as an internal standard, and the plasma concentrations of astragaloside Ⅳ at different time points were determined by UPLC-MS/MS. The chromatographic separation was performed on a Waters Acquity UPLC BEH-C_(18) column(2.1 mm×100 mm, 1.7 μm) via gradient elution. The mobile phase was acetonitrile(A) and 5 mmol·L~(-1) ammonium formate solution with 0.2% formic acid(B). The flow rate was 0.4 mL·min~(-1), the injection volume 5 μL and the column temperature 40 ℃. The mass spectrometry was carried out with electrospray ionization source(ESI) in multiple reaction monitoring and positive ion modes. The specificity, linearity range, accuracy, precision, stability and dilution effect of the method all met the requirements for the determination of astragaloside Ⅳ in plasma. Plasma concentration-time curves were plotted and relevant pharmacokinetic parameters were calculated by DAS 3.2.8. The results showed that the concentration of absorbed astragaloside Ⅳ increased within 0-3.95 h and began to decline since 3.95 h. After 36 h, the metabolism was complete. The area under the plasma concentration-time curve(AUC_(0-t)) and the peak concentration(C_(max)) of astragaloside Ⅳ were increased in the three administration groups compared with the model group, but without significant difference, which suggested that the pharmacokinetic characteristics of saponin components would not necessarily change after the drug-induced alteration of intestinal flora.


Subject(s)
Animals , Chromatography, High Pressure Liquid , Chromatography, Liquid , Diabetes Mellitus, Type 2 , Gastrointestinal Microbiome , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Saponins , Tandem Mass Spectrometry , Triterpenes
7.
Article in Chinese | WPRIM | ID: wpr-906764

ABSTRACT

@#A deuterated internal standard quantitative analysis method based on liquid-liquid extraction-ultra performance liquid chromatography-tandem mass spectrometry (LLE-UPLC-MS/MS) for simultaneous determination of 10 illicit drugs in wastewater was established.Wastewater samples were concentrated by liquid-liquid extraction with dichloromethane: ethyl acetate (1∶1), and separated by a linear gradient of 0.1% formic acid-5 mmol/L ammonium formate aqueous solution and acetonitrile on a C18 column. The samples were then detected by ESI positive ion mode and multiple reaction monitoring mode (MRM) for quantitative analysis.All analytes had a good linear relationship (r ≥ 0.995 7) within the range of their respective standard curves; the limit of quantification was 1 ng/L (except amphetamine at 2.5 ng/L); the relative recovery rate ranged from 96.36% to 106.43%, and the intra- and inter-day precisions were less than 4.70%.This method is accurate, reliable and reproducible, and is suitable for the quantitative determination of 10 illicit drugs in wastewater.It is also suitable for wastewater with complex matrixes that affect solid phase extraction and enrichment.It provides a new analytical method for real-time monitoring of drug abuse.

8.
Article in Chinese | WPRIM | ID: wpr-906496

ABSTRACT

Objective:To explore the difference in antibacterial mechanism between <italic>Coptis chinensis</italic> and<italic> </italic>its<italic> </italic>flower stalk based on secondary metabolites and network pharmacology. Method:Based on the ultraperformance liquid chromatography mass spectrometry(UPLC-MS/MS) detection platform,the secondary metabolites database of <italic>C. chinensis</italic> and its flower stalk(MWDB) was built. The common database of metabolites information and the multivariate statistical analysis were used to study the differences of secondary metabolites between <italic>C. chinensis</italic> and its flower stalk and screen out 18 metabolites of<italic> </italic>the<italic> </italic>flower stalk and 11 metabolites of <italic>C. chinensis</italic> with a high content. BATMAN-TCM database was used to obtain the targets of component action,and their corresponding genes were inquired in the UniProt database. GeneCards was retrieved for antimicrobial genes,and the intersection genes of components and antimicrobials were obtained on Venny platform. Through DAVID gene ontology(GO) enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis,the mechanism of its action was predicted,and the results were visualized through histogram and advanced bubble diagram drawn by GraphPad Prism software and OmicShare database. The protein-protein interaction(PPI) network was constructed by STRING, database and the component-target-pathway network was constructed by Cytoscape 3.7.2 software. The antibacterial differences were compared based on the results of network pharmacology analysis. Result:Through network pharmacology,the antibacterial active components of <italic>C. chinensis</italic> were 5 fewer than that of the flower stalk,55 more antibacterial targets than that of the flower stalk; quercetin and berberine were predicted to be the common components of the antagonistic action of <italic>C. chinensis </italic>and the flower stalk. Key genes involved in antimicrobial action were p38 Mitogen-activated protein kinase 14(MAPK14),catalase(CAT); malaria and Toll-like receptor signaling pathway were different key pathways involved in antimicrobial activity. Conclusion:<italic>C. chinensis </italic>and the flower stalk mainly exert the antibacterial effect in a multi-target and multi-pathway manner,which can offer new ideas and clues for the study of antibacterial mechanism of<italic> C. chinensis</italic> and the flower stalk,and provide a new development direction for the comprehensive development and rational application of the flower stalk resources.

9.
Article in Chinese | WPRIM | ID: wpr-906435

ABSTRACT

Objective:To compare the regulatory effects of <italic>Polygala tenuifolia</italic> and licorice-simmered <italic>P. tenuifolia</italic> on learning and memory, hypothalamus-pituitary-adrenal axis (HPA axis) function and neurotransmitters in rats with heart-kidney imbalance insomnia. Method:The rat model of insomnia induced by multi-factor stimulation was established. After the model being made, the administration groups were given the extracts of <italic>P. tenuifolia</italic> and licorice-simmered <italic>P. tenuifolia</italic> by gavage (dose of 8 g·kg<sup>-1</sup>·d<sup>-1</sup>), while the normal group and model group were given the same volume of normal saline for 7 days. Morris water maze was used to detect the changes of learning and memory ability of rats in each group. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortisol (CORT) in serum of rats from each group. The contents of 5-hydroxytryptamine (5-HT), dopamine (DA), <italic>γ</italic>-aminobutyric acid (GABA) and glutamic acid (Glu) in the hypothalamus of rats were determined simultaneously by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Result:Compared with the normal group, the spatial learning and memory ability of rats in the model group<italic> </italic>was decreased, the times and time of staying in target quadrant were significantly reduced (<italic>P<</italic>0.05, <italic>P<</italic>0.01), the levels of CORT, CRH and ACTH in serum were significantly increased (<italic>P<</italic>0.01), the contents of GABA, DA, 5-HT in hypothalamus tissue were significantly decreased (<italic>P<</italic>0.01), and the content of Glu was significantly increased (<italic>P<</italic>0.01). Compared with the model group, the spatial learning and memory ability of rats in the <italic>P. tenuifolia</italic> group and licorice-simmered <italic>P. tenuifolia </italic>group<italic> </italic>were increased, the times and time of staying in the target quadrant were significantly increased (<italic>P<</italic>0.05, <italic>P<</italic>0.01), the levels of CORT, CRH and ACTH in serum were significantly decreased (<italic>P<</italic>0.05, <italic>P<</italic>0.01), the contents of GABA, DA and 5-HT in hypothalamus tissue were significantly increased (<italic>P<</italic>0.05, <italic>P<</italic>0.01), and the content of Glu was significantly decreased (<italic>P<</italic>0.05). The recovery degree of each index in the licorice-simmered <italic>P. tenuifolia</italic> group was better than that in the <italic>P. tenuifolia</italic> group. Conclusion:Both <italic>P. tenuifolia</italic> and licorice-simmered <italic>P. tenuifolia</italic> can improve the learning and memory ability, improve the function of HPA axis, regulate the level of central neurotransmitters, and have the effect of calming the mind and improving the intelligence of rats with heart-kidney imbalance insomnia. The effect of licorice-simmered <italic>P. tenuifolia</italic> is better than that of <italic>P. tenuifolia.</italic>

10.
Article in Chinese | WPRIM | ID: wpr-906093

ABSTRACT

Objective:To establish a method for the determination of artemisinin and arteannuin B in different <italic>Artemisia annua</italic> germplasms, compare the differences of the two compounds among different <italic>A. annua</italic> germplasm under the condition of hydroponic homogenization and explore the key factors affecting contents of principal compounds in different<italic> A. annua</italic> germplasms. Method:Seedlings from different <italic>A. annua</italic> germplasms were arranged randomly and fed in a hydroponic cultivation system. Contents of artemisinin and arteannuin B were detected by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) with multi-reaction monitoring mode and ACQUITY UPLC<sup>®</sup> BEH C<sub>18</sub> column (2.1 mm×100 mm, 1.7 μm), mobile phase was water-acetonitrile (95∶5, containing 0.1% formic acid, A) and acetonitrile-water (95∶5, containing 0.1% formic acid, B) for gradient elution (0-3.5 min, 25%-1%A; 3.5-3.6 min, 1%-25%A; 3.6-5.0 min, 25%A), the flow rate was set at 0.4 mL·min<sup>-1</sup>. The content differences of artemisinin and arteannuin B in different provenances of <italic>A. annua</italic> were detected and analyzed statistically. Result:The established method had high sensitivity and good separation. A significant difference of artemisinin and arteannuin B contents was observed in different germplasms under the same culture conditions, that is, under the constant temperature of 25 ℃ in hydroponics. The provenance with higher artemisinin content was Yunnan, and the content was 3 810.597 μg·g<sup>-1</sup>. The highest strain of arteannuin B was Shanxi provenance germplasm with the content of 1 691.747 μg·g<sup>-1</sup>. According to the content of artemisinin, the provenances were arranged as follows:Yunnan, Hainan, Hubei, Guangxi, Zhejiang, Shanxi, Beijing, Shandong, Heilongjiang, and Gansu province germplasms. Correlation analysis showed that there was a significant negative correlation between artemisinin content and latitude of <italic>A. annua</italic>, but there was no significant correlation between contents of artemisinin and arteannuin B and longitude. Conclusion:The contents of artemisinin and arteannuin B among different <italic>A. annua</italic> germplasms were significantly different under the same culture environment, and the dominant factors affecting biosynthesis and accumulation of artemisinin and arteannuin B in <italic>A. annua</italic> may be the genetic background, suggesting that germplasm improvement is the key factor to improve the medicinal quality of <italic>A. annua</italic> in subsequent cultivation.

11.
Article in Chinese | WPRIM | ID: wpr-905997

ABSTRACT

Objective:To establish an ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) for determining the plasma concentrations of 10 active ingredients in Wujiwan at different time points after oral administration, and to compare the pharmacokinetic characteristics between normal rats and rats with chronic visceral hypersensitive irritable bowel syndrome (CVH-IBS). Method:CVH-IBS rat model was prepared by the neonatal rat colon percutaneous transluminal coronary angioplasty (PTCA) balloon stimulation method. After intragastric administration of Wujiwan (0.245 g·kg<sup>-1</sup>), blood was collected from the jugular vein at different time points, and the plasma concentrations of 10 active ingredients (berberine hydrochloride, palmatine hydrochloride, coptisine hydrochloride, jatrorrhizine hydrochloride, epiberberine, dihydroberberine, evodiamine, evodine, paeoniflorin, albiflorin) in Wujiwan was detected simultaneously by UPLC-MS/MS, the pharmacokinetic parameters of each component in normal rats and CVH-IBS rats were calculated. Result:The established UPLC-MS/MS could sensitively and accurately detect the plasma concentrations of 10 active ingredients of Wujiwan in rats. Compared with the normal group, the absorption rates of these 10 active ingredients of Wujiwan in the blood of CVH-IBS rats all decreased to a certain extent, and the peak time (<italic>t</italic><sub>max</sub>) was prolonged. Among them, the <italic>t</italic><sub>max</sub> of berberine hydrochloride and jatrorrhizine hydrochloride were significantly prolonged from 54 minute and 39 minute to 90 minute, respectively (<italic>P</italic><0.05, <italic>P</italic><0.01). Area under the plasma concentration-time curve (AUC<sub>0-</sub><italic><sub>t</sub></italic>) of each component increased, and evodiamine and paeoniflorin were significantly different (<italic>P</italic><0.05,<italic> P</italic><0.01). The clearance rates (CL/<italic>F</italic>) of these 10 active ingredients were all decreased, among which berberine hydrochloride, palmatine hydrochloride and evodiamine had significant differences (<italic>P</italic><0.05, <italic>P</italic><0.01). Conclusion:There are significant differences in the pharmacokinetic behavior of the active ingredients in Wujiwan between normal rats and CVH-IBS rats, which may be related to the destruction of microstructure of intestinal epithelial cells and the change of activity of liver enzymes under the pathological state of IBS.

12.
Article in Chinese | WPRIM | ID: wpr-905908

ABSTRACT

Objective:To establish ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for simultaneous determination of six hepatotoxic pyrrolizidine alkaloids in Verbenae Herba, and to carry out preliminary risk assessment according to the research results. Method:An ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm) was used for analysis with 0.05% formic acid and 2.5 mmol·L<sup>-1</sup> ammonium formate in water (A)-0.05% formic acid and 2.5 mmol·L<sup>-1</sup> ammonium formate in acetonitrile (B) as mobile phase for gradient elution (0-12 min, 3%-8%B; 12-25 min, 8%-15%B; 25-26 min, 15%-3%B; 26-30 min, 3%B), the flow rate was 0.3 mL·min<sup>-1</sup>, the column temperature was 40 ℃, and the injection volume was 1 μL. MS system was operated by electrospray ionization (ESI) in the positive ion mode with multiple reaction monitoring mode. MS parameters of triple quadrupole and six analytes were optimized for qualitative and quantitative analysis. According to the determination results, the risk assessment was carried out by using margin of exposure (MOE) combined with transfer rate of hot water extraction. Result:Based on the instrument precision, linear range, repeatability, stability, recovery and other methodological validations, the results were in conformity with relevant standards of quantitative analysis. The linear ranges of intermedine, lycopsamine, intermedine <italic>N</italic>-oxide, lycopsamine<italic> N</italic>-oxide, echimidine<italic> N</italic>-oxide and echimidine were good (<italic>r</italic>≥0.999 0) between peak area and mass concentration in the ranges of 0.984-49.20, 0.994-49.70, 1.012-50.60, 1.032-51.60, 1.004-50.20, 1.016-50.80 µg·L<sup>-1</sup>, respectively. The average recoveries of these six analytes were 87.2%-94.2% with relative standard deviation (RSD)<4.0%. Their MOE values were >10 000. Conclusion:The UPLC-MS/MS established in this study is stable and feasible, which can provide scientific basis for the quality control and safety evaluation of hepatotoxic pyrrolizidine alkaloids in Verbenae Herba.

13.
China Pharmacy ; (12): 2478-2484, 2021.
Article in Chinese | WPRIM | ID: wpr-887428

ABSTRACT

OBJECTIVE:To compare the c hemical components differences of Inula japonica before and after honey-frying. METHODS:UPLC-MS/MS method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C 18 column with mobile phase consisted of 0.1% formic acid-acetonitrile (gradient elution )at the flow rate of 0.3 mL/min. The column temperature was set at 30 ℃,and sample size was 5 µL. The electrospray ion source was scanned by positive ion mode. The first order mass spectrometry scanning range was m/z 70-1 050,the second order mass spectrometry scanning range was m/z 50-1 050, and the normalized collision energy was 40,60 eV ;mass spectrum type was the peak figure ,the flow rate of sheath gas was 35 arb,the auxiliary airflow speed was 10 arb,the spray voltage was 3.80 kV,the S-lens voltage was 50 V,the heating temperature was 350 ℃,and the capillary temperature was 350 ℃. The components were identified by Qual Browser 4.1.39.1 software, referring to the online high-resolution database mzCloud and local database OTCML of high-resolution mass spectrometry of TCM , and combined with relevant literature. The principal component analysis (PCA)and orthogonal partial least squared-discriminant analysis(OPLS-DA)of I. japonica before and after honey-fried were performed by using SIMCA 14.1 statistical software ,and variable importance projection (VIP)value greater than 1 was used as the standard to screen the differential components before and after honey-frying. RESULTS :A total of 29 common chemical components were identified from I. japonica and honey-fried I. japonica,including 5 phenolic acids as 1-caffeoylquinic acid ,chlorogenic acid and 3,5-dicaffeoylquinic acid ,12 flavonoids as quercetin,luteolin and evamectin ,as well as 12 sesquiterpene lactones as 1-O-acetylinula diester ,inula bicolor lactone B and 1-O-acetyl-6-O-isobutyryl inulin. The results of PCA showed that I. japonica and honey-fried I. japonica were located on both sides of the score diagram respectively. The results of OPLS-DA showed that the VIP values of 7 components were greater than 1,which were peak 19(britanin),peak 6(quercetagitrin),peak 1(1-caffeoylquinic acid ),peak 21(vitexicarpin),peak 20(tomentosin), peak 13(spinacetin)and peak 3(daphnetin). CONCLUSIONS :After honey-fried ,the content of chemical components of I. japonica changed and decreased to a certain extent. Britanin ,quercetagitrin,1-caffeoylquinic acid ,tomentosin,vitexicarpin, spinacetin and daphnetin may be the differential components of I. japonica and honey-fried I. japonica .

14.
Acta Pharmaceutica Sinica ; (12): 1439-1443, 2021.
Article in Chinese | WPRIM | ID: wpr-887081

ABSTRACT

An UPLC-MS/MS method was established for the quantification of the genotoxic impurities bis(2-chloroethyl)amine hydrochloride and 1-(3-chloropropyl)-4-(3-chlorophenyl)piperazine hydrochloride in trazodone hydrochloride. The chromatographic separation of the two genotoxic impurities was performed on Waters ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm) at 20 ℃. A mixture of 5 mmol·L-1 ammonium hydrogen carbonate aqueous solution and acetonitrile at a flow rate of 0.3 mL·min-1 in gradient elution mode was employed as mobile phase. The UPLC-MS/MS was equipped with electrospray ionization in positive ionization mode and adopted multiple reaction monitoring mode. We found that the calibration curves of the two genotoxic impurities were linear in the range of 0.1-10 ng·mL-1. The limit of detection was 0.10 ng·mL-1 for bis(2-chloroethyl)amine hydrochloride and the average recovery was 101.53% (RSD = 4.06%). The limit of detection was 0.01 ng·mL-1 for 1-(3-chloropropyl)-4-(3-chlorophenyl)piperazine hydrochloride and the average recovery was 97.95% (RSD = 1.27%). The sample solution was stable for 24 h. No bis(2-chloroethyl)amine hydrochloride was detected in the samples, and the content of 1-(3-chloropropyl)-4-(3-chlorophenyl)piperazine hydrochloride in the samples was within the limit. This research provides a method to improve the quality control standards of trazadone.

15.
Acta Pharmaceutica Sinica ; (12): 2389-2393, 2021.
Article in Chinese | WPRIM | ID: wpr-886967

ABSTRACT

A fast and sensitive UPLC-MS/MS method was established for the direct quantification of insulin aspart in human plasma. The plasma samples were extracted by solid phase extraction (SPE), an ESI ion source was used and operated in the positive ion mode with multiple reaction monitoring (MRM). Bovine insulin was chosen as internal standard and the chromatographic separation of insulin aspart was performed on Waters ACQUITY UPLC CSH C18 column (50 mm×2.1 mm, 1.7 μm). A mixture of acetic acid aqueous solution and acetonitrile with acetic acid at a flow rate of 0.6 mL·min-1 in gradient elution mode was employed as mobile phase. We found that the method was validated over the range of 0.200-10.0 ng·mL-1 for insulin aspart and showed excellent linearity. The intra-and inter-assay accuracy and precision were below 14.5% and the recovery was 36.7%-41.7% over the three concentration levels evaluated. The UPLC-MS/MS method was selective, accurate, sensitive and robust, and the method was successfully applied in supporting the pharmacokinetic research of two insulin aspart injections (Test Product and NovoRapid®) in heathy male subjects. This clinical trial was approved according to the Ethics Committee of West China Hospital, Sichuan University (2017 Clinical Trial (Western Medicine) Approval (148)).

16.
Article in Chinese | WPRIM | ID: wpr-886879

ABSTRACT

Objective To develop an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to simultaneously detect the contents of gallic acid, syringin, phellodendrine, aesculin and rhein in the gallnut lotion. Methods An UPLC- MS/MS method was established. Separation was performed on an Agilent Poroshell 120 EC-C18(2.1 mm×150 mm, 2.7 μm)with a gradient mobile phase system of 0.2% formic water-acetonitrile solution. The flow rate was 0.3 ml/min. The temperature of column was 30 ℃. The injection volume was 2 μl. The MS detection was in dynamic MRM mode. Results gallic acid, syringin, phellodendrine, aesculin and rhein were successfully separated using this method, with good linear relationship as the ranges of 153.8-15380、10.31-1031、5.265-526.5、50.70-5070、1.054-105.4 ng/ml, respectively. The precision, repeatability, stability and recovery were good. Conclusion This UPLC-MS/MS method is stable, rapid, and reproducible., It is suitable for detecting the contents of gallic acid, syringin, phellodendrine, esculetin and in the gallnut lotion.

17.
Article in Chinese | WPRIM | ID: wpr-886875

ABSTRACT

Objective To establish an assay method for diphenhydramine hydrochloride and caffeine in rat plasma by UPLC-MS/MS for pharmacokinetic study. Methods The chromatographic separation was performed on an ACE 3 C18-PFP (3.0 mm×150 mm, 3 μm) by isocratic elution with the mobile phase of water containing 0.1% formic acid and acetonitrile (62:38, V/V). MS condition was optimized in the positive ion detection mode by multiple reaction monitoring (MRM), along with the Agilent JetStream electrospray source interface (AJS-ESI). The precursors to the product ion transitions were 256.2→167.0 (m/z) for diphenhydramine hydrochloride, 262.0→167.0 (m/z) for the internal standard (IS) diphenhydramine-D6, 195.0→138.0 (m/z) for caffeine and 204.0→116.2 (m/z) for the IS caffeine-D9. Results The calibration curve was linear in the range of 1-1×103 ng/ml for diphenhydramine hydrochloride in rat plasma (r=0.999 6), and in the range of 15-1.5×105 ng/ml for caffeine in rat plasma, (r=0.999 9). The intra-day and inter-day precision and accuracy were good (RSD<10%, RE<±10%). Pharmacokinetic studies showed that metabolic characteristics of diphenhydramine hydrochloride 10-30 mg/kg and caffeine 24-72 mg/kg were linear after intragastric administration. The two components were metabolized in rats with gender difference, the cmax and the AUC of diphenhydramine hydrochloride and caffeine were greater in female than those in males. Conclusion This method is accurate, rapid and sensitive. It can be used for the determination of diphenhydramine hydrochloride and caffeine in rat plasma collected for pharmacokinetic study. The results of pharmacokinetic studies in rats provide reliable data support for the clinical application of the compound preparation.

18.
China Pharmacy ; (12): 2248-2253, 2021.
Article in Chinese | WPRIM | ID: wpr-886807

ABSTRACT

OBJECTIVE :To est ablish the method for the determination of 20(S)-protopanaxadiol(PPD)concentration in human plasma. METHODS :Plasma samples were precipitated with acetonitrile and determined by UPLC-MS/MS ,using finandrogen as internal standard. The determination was performed on Waters ACQUITY UPLC HSS T 3 column with mobile phase consisted of 5 mmol/L ammonium bicarbonate aqueous solution-acetonitrile (gradient elution )at the flow rate of 0.4 mL/min. The column temperature was set at 40 ℃,and sample size was 10 μL. The ion source was electrospray ion source,and negative ion scanning was carried out with multiple reaction monitoring mode . The ion pairs used for quantitative analysis were m/z 459.40→ 375.20(PPD)and m/z 371.30→315.30(internal standard ). At the same time ,the method was applied to the determination of clinical samples. RESULTS :The linear range of PPD was 0.25-30.00 ng/mL(r=0.999 2),and the limit of quantitation was 0.25 ng/mL. RSDs of intra-batch and inter-batch were all lower than 10%,and relative errors (RE)were -14.61%-12.69%. Extraction method and matrix effect did not affect the quantitative determination of PPD. In ginsenoside CK 100 mg group ,ginsenoside CK 200 mg group and ginsenoside CK 300 mg group ,mean cmax of patients with rheumatoid arthritis after oral administration of corresponding drugs were 18.06,30.03,27.00 ng/mL;median tmax were 12.0,6.0,12.0 h;mean AUC 0-t were 622.52,668.15, 1 155.97 ng·h/mL. CONCLUTIONS :The method for the determination of PPD concentration in human plasma is successfully established. The method is sensitive ,accurate, kq1907011) stable,easy to operate and less plasma consumption. It can be used for the quantitative determination of clinical samples.

19.
Organ Transplantation ; (6): 595-2021.
Article in Chinese | WPRIM | ID: wpr-886789

ABSTRACT

Objective To establish a detection system of ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for everolimus concentration in whole blood of liver transplant recipients. Methods The proteins of samples were precipitated with methanol and zinc sulfate, and everolimus-D4 was used as the internal standard. Phenomenex Kinetex PFP column was used. The mobile phase A was water (containing 2 mmol/Lammonium formate and 0.1% formic acid), and the mobile phase B was methanol (containing 2 mmol/L ammonium formate and 0.1% formic acid). The gradient elution was performed with the flow rate of 1 mL/min, the column temperature of 50 ℃ and the injection volume of 1 μL. The multi-reaction monitoring mode was used to quantitatively analyze with electrospray positive ionization. The UPLC-MS/MS detection system required only 100 μL of whole blood, and could achieve a sufficient lower limit of quantification without complicated sample preparation. The total running time was within 4.5 min. Linear regression (1/x2) analysis was performed using peak area of everolimus / peak area of everolimus-D4 (y) and concentration of everolimus/concentration of everolimus-D4 (x) to calculate the calibration function and analyze its accuracy and linear relationship. UPLC-MS/MS was used to detect the trough blood concentration of everolimus in blood samples of 5 recipients after liver transplantation. Results The accuracy of quality control was within 15%, and the linear relationship of everolimus was good in the blood concentration range of 1-100 ng /mL(R2 > 0.990). Trough blood concentration of everolimus measured in blood samples of 5 liver transplant recipients ranged from 3.77 to 9.27 ng/mL. Conclusions The detection system of UPLC-MS/MS in this study is suitable for monitoring the concentration of everolimus in whole blood of liver transplant recipients because of its high accuracy, simple sample processing method and short detection time.

20.
China Pharmacy ; (12): 1728-1733, 2021.
Article in Chinese | WPRIM | ID: wpr-882144

ABSTRACT

OBJECTIVE:To establish the method for determining protein binding rate of dipeptidyl peptidase- 4 inhibitor LGT- 6 in different species of plasma ,and to compare their difference. METHODS :By equilibrium dialysis ,LGT-6(3,30,300,3 000 nmol/L)was equilibrated in rat ,monkey and human plasma (i. e. internal dialysis solution )for 48 h,using phosphate buffer as the external dialysis solution. The concentration of LGT- 6 in internal and external dialysis solution was determined by UPLC-MS/MS using tolbutamide as internal standard ,and the plasma protein binding rate was calculated. The determination was performed on ACQUITY UPLC HSS T 3 column with water (containing 0.01% formic acid )-acetonitrile(containing 0.01% formic acid )as mobile phase at the flow rate of 0.6 mL/min. The column temperature was 40 ℃,and the sample size was 2 μL. The ion source was electrospray ion source ,and the multiple ion monitoring mode was used to carry out positive ionization scanning. The ion pairs for quantitative analysis were m/z 487.0→434.3(LGT-6),m/z 271.1→172.0(internal standard ),respectively. RESULTS :At the concentrations of 3,30,300,and 3 000 nmol/L,the protein binding rates of LGT- 6 in rat plasma were (96.25±0.97)%,(84.16± 1.24)%,(78.25±0.61)%,(66.63±0.95)%;the protein protein binding rates in monkey plasma were (98.54±0.58)%,(87.27± 1.01)%,(79.35±0.86)%,(66.69±0.54)%;the protein binding rates in human plasma were (99.40±1.03)%,(84.48± 1.15)%,(77.62±0.77)%,(66.93±0.48)%. At the same concentration ,the protein binding rates of LGT- 6 in rat ,monkey and human plasma had no significant difference (P>0.05). In the same species of plasma ,there were significant differences in the plasma protein binding rates of different concentration of LGT-6 among those groups (P<0.05),and it decreased with 才〔2016〕4015) the increase of drug concentration. CONCLUSIONS : The method for the determination of plasma protein binding rate of LGT-6 is successfully established. The data revealed that the protein binding rate of LGT- 6 is concentration-dependent , there was no obvious spec ies difference on protein binding rates of LGT- 6 in rat ,monkey and human plasma under the same concentration.

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