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ObjectiveIn order to explore the changes of chemical constituents in Plantaginis Semen before and after stir-frying， ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MSE） was used to rapidly identify and semi-quantitatively analyze the differential components in Plantaginis Semen processed at different stir-frying time. MethodWaters ACQUITY UPLC BEH C18 column （2.1 mm×100 mm， 1.8 μm） was employed with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-1 min， 5%-10%B； 1-2 min， 10%-15%B； 2-10 min， 15%-20%B； 10-12 min， 20%-40%B； 12-13 min， 40%-100%B； 13-14 min， 100%-5%B； 14-15 min， 5%B）， the flow rate was 0.3 mL·min-1， the column temperature was 40 ℃， and the injection volume was 3 μL. Electrospray ionization （ESI） was applied for mass spectrometric analysis under positive and negative ion modes， and the scanning range was m/z 50-1 500. MarkerLynx 4.1 software was used to find the differential compounds， and the intensity of each ion peak in samples with different stir-frying time was compared to study the content variations of these compounds. ResultA total of 20 components with potential significant differences were found， among which 17 were identified and 3 were unknown， mainly including phenylethanoid glycosides， iridoid glycosides， alkaloids and others. After processing， the peak intensities of 7 compounds， such as sucrose， geniposidic acid， verbascoside and plantagoguanidinic acid A， in Plantaginis Semen decreased. The peak intensities of orobanchoside， dianthoside and plantain D increased first and then decreased during the stir-frying process. The peak intensities of 10 compounds （decaffeoylacteoside， calceolarioside A， isoacteoside， etc.） increased， and 9 of them were newly generated components. ConclusionThe content and composition of the chemical components in Plantaginis Semen changed significantly after stir-frying， which may be related to the reduction of laxative effect and the enhancement of antidiarrheal and diuretic activities of Plantaginis Semen after stir-frying.
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Objective:In order to systematically clarify the chemical composition of Jiechangyan Qixiao granules， the main chemical components in this preparation were rapidly identified and assigned by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MS<sup>E</sup>）. Method:ACQUITY UPLC BEH C<sub>18</sub> column （2.1 mm×100 mm， 1.8 μm） was employed for UPLC analysis with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-2 min， 5%B； 2-16 min， 5%-21%B； 16-30 min， 21%-95%B； 30-33 min， 95%B； 33-34 min， 95%-5%B； 34-37 min， 5%B）. The flow rate was 0.3 mL·min<sup>-1</sup>， the column temperature was 30 ℃， and the volume of sample injection was 2 μL. Electrospray ionization （ESI） was applied for scanning under positive and negative ion modes with the scanning range of <italic>m</italic>/<italic>z</italic> 60-1 200. MS<sup>E</sup> mode was used to collect mass spectral data. The ion peaks were identified by comparing with the information of control substances， literature references and self-built database. Result:A total of 102 chemical components were separated and identified in Jiechangyan Qixiao granules， including organic acids， flavonoids and its glycosides， triterpenes， phenylethanoid glycosides， tannins， iridoid glycosides and other components， among which flavonoids and its glycosides were from Drynariae Rhizoma and Crataegi Fructus， phenylethanoid glycosides and iridoid glycosides were from Plantaginis Semen， triterpenoids and tannins were from Crataegi Fructus and Chebulae Fructus. Among the identified chemical constituents， there were 28 from Drynariae Rhizoma， 31 from Plantaginis Semen， 53 from Chebulae Fructus and 58 ingredients from Crataegi Fructus. Conclusion:The established UPLC-Q-TOF/MS<sup>E</sup> can comprehensively and rapidly analyze the chemical constituents in Jiechangyan Qixiao granules， and preliminarily elucidates the chemical composition profile of this granules， which can lay a foundation for further research on the pharmacodynamic material basis and quality control of Jiechangyan Qixiao granules.
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Objective: To realize the classification and identification of Cynomorii Herba from different producing areas based on fourier transform infrared spectroscopy (FTIR) and chemometrics. Method: FTIR spectrum data of 106 batches of Cynomorii Herba from 12 cities in 5 provinces were collected by transmission method and preprocessed. The FTIR fingerprints of Cynomorii Herba were established, and spectrum analysis was performed. The FTIR similarities of Cynomorii Herba from different producing areas were calculated by correlation coefficient method. The first derivative (1D) spectrum of average FTIR of Cynomorii Herba from different producing areas were obtained. The soft independent modeling of class analog (SIMCA) model based on principal component analysis (PCA) was established by the preprocessed 1D spectrum data. The orthogonal partial least squares (OPLS) model was established by top 6 principal components. Result: The FTIR fingerprint trend and main absorption peaks of Cynomorii Herba from different producing areas were basically the same,and 16 common characteristic absorption peaks were recognized. Similarity and 1D spectrum of FTIR fingerprint of Cynomorii Herba from different producing areas showed significant and unique characteristics. The established SIMCA model can realize the classification and identification of Cynomorii Herba from different provinces,while OPLS model can realize accurate classification and identification of Cynomorii Herba in different cities. The classification and identification of Cynomorii Herba from 12 city producing areas showed obvious geographical clustering characteristics. Conclusion: The established method based on FTIR and chemometrics can realize the classification and identification of Cynomorii Herba from 12 cities.
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Ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to establish the chromatography fingerprint for aerial parts of Angelica sinenis(AAS) from 10 different places. Acetyl-phenyl-hydrazine(APH) was used to duplicate the mouse model of blood deficiency. Then partial least squares regression was used to investigate the spectrum-effect relationship between the relative contents and the data of enriching blood pharmacodynamics efficacy. The results showed that the three groups of high, medium and low doses of AAS had certain enriching blood activities(P<0.05), and the high dose group had the best effect(P<0.01). The contribution degree of the AAS to enriching blood activities of each common peaks were determined by PLS regression coefficient. Among them, 7 common peaks, including P17(unknown), P18(unknown), P19(unknown), P28(alisol B 23-acetate or its isomer), N5(luteolin), N11(1-caffeoylquinicacid,1-O-caffeoylquinic acid) and N14(unknown), contributed significantly to the effect of enriching blood activities. This paper dealed with the investigation on the spectrum-effect relationship between enriching blood activities and LC-MS chromatography fingerprint of AAS, and determination of the effective compositions of AAS with enriching blood activities. It provided theoretical foundation for the comprehensive development and utilization of AAS.
Subject(s)
Animals , Mice , Angelica , Chemistry , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacology , Mass Spectrometry , Medicine, Chinese Traditional , Plant Components, Aerial , ChemistryABSTRACT
Ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(LC-MS) was used to establish the chromatography fingerprint for fresh(FRAS) and dry(RAS) roots of Angelica sinensis from 10 different places. The rat model of blood deficiency was established by acetyl-phenyl-hydrazine(APH) and cyclophosphamide(CTX). Then grey relational analysis(GRA) and partial least squares regression(PLS) were used to investigate the spectrum-effect relationship between the relative contents and the data of enriching blood pharmacodynamics efficacy. The results showed that the FRAS and RAS had certain enriching blood activities(P<0.05). The contribution degree of the FRAS and RAS to enriching blood activities of each common peaks were determined by regression coefficient. Among them, 4 common peaks contributed significantly to the effect of enriching blood activities, P1(unknown), P2(unknown), P7(ferulic acid), and P11(senkyunolide A) respectively. This paper investigated the spectrum-effect relationship between enriching blood activities and LC-MS chromatography fingerprint of RAS and FRAS, and determined the effective compositions of RAS and FRAS with enriching blood activities. It lays a theoretical foundation for the comprehensive development and utilization of A. sinensis.
Subject(s)
Animals , Rats , Angelica sinensis , Chemistry , Chromatography, Liquid , Drugs, Chinese Herbal , Pharmacology , Mass Spectrometry , Phytochemicals , Pharmacology , Plant Roots , ChemistryABSTRACT
Objective To explore the relationship between the volatile components in Angelicae Sinensis Radix from different regions of Gansu Province and its growing environment with metabolomics based on GC-MS. Methods The GC-MS method was used for detecting the volatile components in Angelicae Sinensis Radix from 31 different regions in Gansu province, and principal component analysis (PCA) and partial least squares (PLS) methods were used for analyzing and evaluating its relationship with the growing environment. Results The results of PCA showed that the volatile components in Angelicae Sinensis Radix from different regions in Gansu province were related to the altitude and the soil types. The PLS method could divide 31 samples of Angelicae Sinensis Radix from different regions in Gansu Province into three groups according to the difference of altitude. There were significant differences in the volatile components in the samples taken at different altitude regions. After analyzing linear loading plots from PCA and PLS, 11 charateristic components were screened out, including 7 compounds were identified by the retrieval of NIST11 database. Conclusion The volatile components in Angelicae Sinensis Radix from different regions in Gansu Province are closely related to the altitude and the soil type.
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<p><b>OBJECTIVE</b>To investigate the expression level of WT1 gene in bone marrow of patients with acute myeloid leukemia (AML) and its relationship with prognosis.</p><p><b>METHODS</b>The copy numbers of WT1 and internal reference gene in bone marrow samples from 75 newly diagnosed AML patients were detected by using real-time quantitative PCR. The gene WT1 expression level was determined by the ratio of the copy numbers of WT1 to reference gene. And the clinical characteristics, the complete remission (CR) rate after induction chemotherapy, 2-year overall survival (OS) rate and event-free survival (EFS) rate were calculated and analysed.</p><p><b>RESULTS</b>The expression level of WT1 did not significantly correlate with common clinical parameters such as age, sex, molecular abnormality, FAB classification and risk stratification. The CR rate in the high WT1 expression group before treatment was 65.4%, which was lower than that of 93.9% in the low expression group (χ2=8.25, P<0.01). The 2-year overall survival rate and event-free survival rate of the two groups were statistically significantly different (P<0.05), and the OS and EFS rates in high WT1 expression group were lower than those in low expression group. After the induction chamotheropy for about 1, 3 month and 6 months, the 2-year OS rate significantly increased in patients with decrease of WT1 gene expression level by one log or more (P<0.05).</p><p><b>CONCLUSION</b>The expression level of WT1 gene in bone marrow may be an effective marker to evaluate therapy efficacy and prognosis for AML patients (non APL).</p>
Subject(s)
Humans , Bone Marrow , Metabolism , Disease-Free Survival , Genes, Wilms Tumor , Induction Chemotherapy , Leukemia, Myeloid, Acute , Diagnosis , Genetics , Prognosis , Real-Time Polymerase Chain Reaction , Remission Induction , Survival Rate , WT1 Proteins , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the features of immunophenotypes and the characteristics of molecular biology and cellular genetics of AML patients with CD7 and CD4 expression.</p><p><b>METHODS</b>The immunophenotypical markers of AML cells were detected by multiple parameter flow cytometry; the expression of WT1, MDK, ETO, PML-RaRa and BCR-ABL were detected by RT-PCR; and cellular features were analyzed by R-band in 304 patients. The patients were divided into three groups according to their immunophenotypes: AML with CD7 expression (CD7 group), AML with CD4 expression(CD4 group) and AML without CD7 and CD4 expression (common AML group).</p><p><b>RESULTS</b>The expression rate and level of HLA-DR in CD7 group were higher than those in the common AML group, and the expression rate of CD33 and CD34 was higher than that in the other two groups. The expression rate and level of CD15, CD64 in the CD4 group were higher than those in the other 2 groups, and the expression rate and level of CD33 were higher than those in the common AML group. WT1 expression in the CD7 group was lower than that in the common AML group. PML-RaRa was not detected in the CD7 group. AML with co-expression of CD4 or CD7 showed more normal karyotype. (15;17) was not found in AML with CD7 expression.</p><p><b>CONCLUSION</b>AML cells with CD7 expression originate from precursor cells and are blocked in the early phase of hematological development; AML cells with CD4 expression originate from more mature stage of hematological devevelopment and with CD33, CD64 and CD15 high expression; AML cells with CD7 and CD4 expression are characterized by no-specific change of cellular genetics. According to the expression level and intesity of CD4 and CD7, and together with other specific lineage markers, the MRD in AML patients can be quantitatively detected.</p>