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Objective: To establish an HPLC fingerprint spectrum of for decoction pieces-, standard decoction- and dispensing granules of Astragali Radix, and evaluate the correlation among them. Methods: HPLC method was used to determine the content of 7-hydroxy- 4'-methoxy isoflavone, calycosin-7-glucoside, calycosin, and ononin in decoction pieces-, standard decoction- and dispensing granules of Astragali Radix. The common pattern of the three fingerprints was established and the correlation was evaluated through the control fingerprint spectrum. Results: The average contents of 7-hydroxy-4'-methoxy isoflavone, calycosin-7-glucoside, calycosin, and ononin in 20 batches of decoction pieces were 0.097, 0.482, 0.142, and 0.237 mg/g, respectively; The average content of these four index components in the standard decoction were 0.116, 0.912, 0.214, and 0.434 mg/g, respectively; The average content of these four index components in the dispensing granules were 0.088,, 0.623,, 0.160, and 0.289 mg/g, respectively. There were six common peaks in all fingerprint spectra of 20 batches of decoction pieces-, standard decoction- and dispensing granules of Astragali Radix. The similarity between dispensing granules and standard decoction was 0.980, indicating that the components difference between them was very small. Conclusion: The study establishes an HPLC fingerprint spectrum, and itwhich reflects the whole picture of multi-component of decoction pieces-, standard decoction- and dispensing granules of Astragali Radix. It provides reference value for the identification and quality control of Astragalus dispensing granules of Astragalus Radix.
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Objective: To optimize the infiltration process of Astragalus (Astragalus membranaceus var. mongholicus) medicinal materials by Box-Behnken response surface method. Methods: Based on the HPLC-DAD-ELSD and response surface design method, the qualified rate of decoction pieces, the content of index components and bending inspection were used as comprehensive inspection indicators, and the three factors of infiltration were selected for response surface experimental design to optimize the infiltration process of Astragalus medicinal materials parameter. Results: The best infiltration process was as following: infiltration temperature was 20 ℃, with water addition of 1:0.988 for 6 h. Under this process, the qualified rate of Astragalus pieces was 95.81%, the content of calycosin-7-glucoside was 0.072%, and the content of astragaloside IV was 0.276 %. Combining fingerprint analysis and heat map analysis, the material basis of A. membranaceus var. mongholicus changed during the infiltration process. The infiltration parameters should be strictly controlled during the infiltration process to ensure uniform quality of the pieces. Conclusion: The optimized Astragalus medicinal material infiltration process is stable and feasible with good reproducibility, which can provide a reference for the mass production process development of Astragalus medicinal slices.
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Objective To investigate the effects of calycosin, formononetin, calycosin-7-glucoside and ononin on PC 12 cells differentiation. Methods PC 12 cells were cultured and treated with different concentrations of nerve growth factor (NGF), calycosin, formononetin, calycosin-7-glucoside and ononin for 5 days, once a day, 3 times in a row. The neurite outgrowth of PC 12 cells was observed and the expression of β III-tubulin were measured by immunofluorescence. Results Compared with the vehicle group, neurite outgrowth and the expression of β III tubulin in PC 12 cells had not promoted by calycosin, formononetin, calycosin-7-glucoside and ononin (0.01-10.00 μmol/L). Conclusion PC 12 cells differentiation could not be induced by calycosin, formononetin, calycosin-7-glucoside and ononin.
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Objective: To establish an HPLC fingerprint of methanol extract of Zhixue Tiaojing Granule (ZTG), and determine the content of paeoniflorin and calycosin-7-glucosid. Methods: HPLC analysis was performed on an Thermo UltiMateTM U3000 LC system and carried out at 30 ℃ on a column of Thermo AcclaimTM 120 C18 (250 mm × 4.6 mm, 5 μm). Gradient elution system was composed of water solution (phase A) and acetonitrile (phase B). Detection was performed at the wavelength of 254 nm, the mobile flow rate was 1 mL/min, the column temperature was 30 ℃, and sample volumn was 10 μL. Results: The HPLC fingerprint for 10 batches of ZTG were obtained and the similarities for 10 batches of samples were all over 0.95. Eleven characteristic peaks were found and peaks were from Astragali Radix (peak 1, 5, 7, 10), Codonopsis Radix (peak 1), Dipsaci Radix (peak 2, 3), Paeoniae Radix Alba (peak 4), Angelicae Sinensis Radix (peak 6), Rubiae Radix et Rhizoma (peak 6, 8, 11), and Agrimoniae Herba (peak 9). Besides, three peaks were assigned include paeoniflorin, calycosin-7-glucosid, and formononetin. The content of paeoniflorin and calycosin-7-glucosid was 1 062 μg/g and 84.08 μg/g. Conclusion: The method can be used for the quality control of ZTG with good precision, accuracy, and reproducibility.
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Objective: A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed to study the pharmacokinetic behaviors of astragaloside, calycosin-7-glucoside, and formononetin in rat plasma after oral administration of Qili Qiangxin Capsule. Methods: SD rats were given 1.3 g/kg Qili Qiangxin Capsule suspension by gavage. Blood samples were collected from inner canthus at different time points. The concentration of the components was determined by HPLC-MS/MS. The chromatographic column was Agilent Zobrax XDB-C18 (50 mm × 4.6 mm, 3.5 μm); The mobile phase was 0.1% formic acid-water (A) and methanol solution containing 0.1% formic acid (B), gradient elution; And the scanning mode was multi-reflection monitoring (MRM) with positive ion mode. The pharmacokinetic parameters were calculated by DAS 3.0 software fitting. Results: The pharmacokinetic parameters tmax of astragaloside, calycosin-7-glucoside and formononetin in rat plasma were (0.67 ± 0.00), (0.67 ± 0.06), and (0.17 ± 0.00) h, Cmax were (120.46 ± 45.03), (31.49 ± 12.00), and (5.17 ± 1.13) ng/mL, AUC0—t were (320.92 ± 79.10), (103.26 ± 47.63), and (14.70 ± 5.48) h•ng/mL, t1/2 were (5.74 ± 0.78), (6.05 ± 3.34), and (5.07 ± 2.13) h, respectively. Conclusion: The method is proved to be specific and repeatable, which is suitable for determination of astragaloside, calycosin-7-glucoside, and formononetin in rat plasma for pharmacokinetic study.
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Objective: To establish HPLC method for the simultaneous determination of saikosaponin a, naringin, paeoniflorin, calycosin-7-glucoside, tanshinone IIA, cinnamaldehyde, schisandrin, syringin, berberine hydrochloride, chrysophanol and hesperidin in Yigan Yiqi Jieyu Granules (YYJG), and conduct a quality assessment using principal component analysis. Methods: The chromatographic separation was achieved on an Caprisil AQ-C18 (150 mm × 4.6 mm, 5.0 μm) column with mobile phase consisted of 0.1% phosphate-acetonitrile for gradient elution, at the flow rate of 0.8 mL/min; The column temperature was 45 ℃. The results of the content were then combined with the principal component analysis to achieve the scientific assessment of the different batches of drugs. Results: The content of saikosaponina, naringin, paeoniflorin, calycosin-7-glucoside, tanshinone IIA, cinnamaldehyde, schisandrin, syringin, berberine hydrochloride, chrysophanol and hesperidin in YYJG had good linear relationship in the ranges of 1.6-80.0, 14-700, 10-500, 1.6-80.0, 1.6-80.0, 2.4-120.0, 1.2-60.0, 1.2-60.0, 8.0-400.0, 2.0-100.0, and 2.0-100.0 μg/mL, respectively; The average sample recovery rate range were 98.3%, 99.2%, 98.8%, 99.3%, 101.9%, 97.5%, 99.8%, 101.7%, 101.1%, 102.5%, and 100.9% (RSD < 2.0%); The content of 11 active ingredients in 16 batches of samples respectively were 0.233-0.322, 3.007-3.142, 2.201-2.273, 0.320-0.355, 0.317-0.399, 0.451-0.523, 0.265-0.297, 0.209-0.226, 1.848-1.873, 0.380-0.425, and 0.615-0.647 mg/g, respectively. Conclusion: The established method is simple, accurate and reproducible, and can provide the reference for the quality control of YYJG.
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Objective: To establish the HPLC fingerprint and determine main components of Qishen Granules (QG), so as to provide a scientific basis for its quality control. Methods: HPLC analysis was performed on an Agilent Eclipse plus C18 column (250 mm × 4.6 mm, 5 μm). The gradient elution was performed by the mobile phase consisting of acetonitrile-0.1% formic acid and 0.1% formic acid aqueous with the flow rate of 1.0 mL/min, the detection wavelength was set at 254 nm, and the column temperature was 30 ℃. Fingerprints of ten batches of QG were determined, and the similarities among fingerprints were evaluated. Attributing analysis of the common peaks was achieved by comparing the retention times with the chromatograms of single constituent drugs, and identifications of common peaks were performed on LC-Q TOF-MS and nine components were further confirmed by the reference substances, the content of the nine compounds was subsequently analyzed by HPLC. Results: The similarities of 10 batches of QG were all greater than 0.991. There were 18 common peaks marked in total, peaks 1, 2, 8, 14 and 18 from Astragalus membranaceus var. mongholicus, peaks 3, 4, 5, 6, 7, 9, 10 and 11 from Lonicera japonica, peaks 12, 13, 15 and 16 from Salvia miltiorrhiza, and peaks 17 and 18 from Glycyrrhiza uralensis. Based on the identification of the common peaks, nine components such as chlorogenic acid (peaks 4), calycosin-7-glucoside (peaks 8), isochlorogenic acid B (peaks 9), isochlorogenic acid A (peaks 10), ononin (peaks 14), salvianolic acid B (peaks 15), salvianolic acid A (peaks 16), glycyrrhizic acid (peaks 17), and formononetin (peaks 18) were identified and quantified. The content of the nine components was determined as 6.676-10.213, 0.628-0.963, 1.018-1.886, 1.082-1.972, 0.477-0.790, 11.327-17.788, 0.519-0.908, 2.000-3.638, and 0.010-0.016 mg/g, respectively. Conclusion: The method established in this study shows good characteristics, specificity, and repeatability, which can provide scientific basis for the quality control of QG.
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Objective:To analyze and identify the brain and blood absorption components of rats after intragastric administration of Buyang Huanwu Tang(BYHWT). Method:The brain tissue,plasma of normal rats and the cerebral ischemia-reperfusion rats were analyzed by UPLC-Q-TOF-MS/MS.The prototype components in BYHWT were identified according to retention time,accurate relative molecular weight,primary and secondary mass spectrometry data. Result:After the administration of BYHWT,five compounds were found to enter the normal brain tissue through the blood-brain barrier and identified as calycosin-7-glucoside,albiflorin,formononetin-7-O-β-D-glucoside-6″-O-acetyl,safflower yellow A and astragaloside A;two compounds penetrated the blood-brain barrier and entered modeling brain tissue,and they were identified as calycosin-7-glucoside and formononetin-7-O-β-D-glucoside-6″-O-acetyl;seven compounds entered normal plasma and were identified as calycosin-7-glucoside,albiflorin,hydroxysafflor yellow A,et al;three compounds entered model plasma and identified as calycosin-7-O-β-D-glucoside-6″-O-acetyl,6″-O-acetyl-(6αR,11αR)-9,10-dimetho-xypterocarpan-3-O-β-D-glucoside and formononetin-7-O-β-D-glucoside-6″-O-acetyl. Conclusion:BYHWT has different pharmacological material basis in normal and cerebral ischemia-reperfusion rats.
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Objective: To establish HPLC coupled with wavelength switching and gradient elution method (HPLC-DVD) for simultaneous determination of ten main components (calycosin-7-glucoside, ruscogenin, amygdalin, ginsenoside Rb1, ginsenoside Re, ferulic acid, crocin I, salvianolic acid B, acetyl-11-keto-β-boswellic acid, and tanshinone IIA) in Shuangshenlong Capsule (SSLC). Methods: The chromatographic separation was achieved on Hypersil ODS C18 (150 mm × 4.0 mm, 3 μm) column with methanol- water (8∶2) T (A)-0.1% phosphoric acid solution (B) as mobile phases for gradient elution, at the flow rate of 0.6 mL/min; The detection wavelength was set at 260 nm for α-calycosin-7-glucoside, 280 nm for ruscogenin, 210 nm for amygdalin, 203 nm for ginsenoside Rb1 and ginsenoside Re, 320 nm for ferulic acid, 440 nm for crocin I, 286 nm for salvianolic acid B, 250 nm for acetyl-11-keto-β-boswellic acid, and 270 nm for tanshinone IIA. The volume of sample injection was 10 μL. Results :The ten active components were well separated and showed good linearity between mass concentration and peak area, such as calycosin-7-glucoside 3.88—69.86 mg/L (r = 0.999 2), ruscogenin 22.1—397.8 mg/L (r = 0.999 1), amygdalin 37.43—673.5 mg/L (r = 0.999 4), ginsenoside Rb1 45.15—812.72 mg/L (r = 0.999 6), ginsenoside Re 4.55—81.95 mg/L (r = 0.999 5), ferulic acid 3.06—55.15 mg/L (r = 0.999 4), crocin I 1.93—34.76 mg/L (r = 0.999 5), salvianolic acid B 15.68—282.15 mg/L (r = 0.999 6), acetyl-11-keto-β-boswellic acid 11.31—203.58 mg/L (r = 0.999 1), and tanshinone IIA 1.89—34.16 mg/L (r = 0.999 6). The precision was good, and RSD was not more than 1.27%. The repeatability was good, and RSD was not more than 1.28%. The stability was good in 8 h, and RSD was not more than 0.96%. The average recoveries and corresponding RSD values were 99.61% (1.21%), 100.11% (0.76%), 101.52% (0.62%), 101.22% (1.03%), 100.83% (1.14%), 98.94% (0.53%), 101.04% (1.09%), 100.05% (1.25%), 99.81% (0.68%), and 101.94% (1.31%), respectively. The contents of nine batches of calycosin-7-glucoside, ruscogenin, amygdalin, ginsenoside Rb1, ginsenoside Re, ferulic acid, crocin I, salvianolic acid B, acetyl-11-keto-β-boswellic acid, and tanshinone IIA were 0.142—0.158, 0.747—0.764, 1.578—1.619, 2.163—2.185, 0.235—0.251, 0.557—0.580, 0.105—0.122, 0.311—0.328, 0.605—0.624, 0.062—0.079 mg/capsule, respectively. Conclusion: HPLC coupled with wavelength switching and gradient elution method has been established for simultaneous determination of ten components in SSLC. The method is simple, quick, accurate, and it can be used for content determination and quality control of SSLC.
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Objective This study aimed to establish an integral quality control method for Naoxintong capsules based on the theory of quality markers (Q-marker) in traditional Chinese medicine via multivariable statistical calculation and "active compounds-targets" network construction. Methods Multi-statistical methods were firstly carried out to explore the specific components from different batches of Naoxintong Capsules. The "Q-markers-targets-pathways-diseases" network was constructed using the Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) online analysis tool, and biological information annotations were made. Moreover, a simultaneously determination method for Q-markers was established by using ultra-high performance liquid chromatography (UPLC). Results Nine compounds were chose as the Naoxintong Q-markers capsules including mulberroside A, hydroxysafflor yellow A, paeoniflorin, ferulic acid, calycosin-7-glucoside, rosemary acid, salvianolic acid B, formononetin, and tanshinone IIA. To verify the prediction, target network construction analysis was performed and indicated that the nine Q-markers mainly targeted five proteins which were PTGIS, PTGS2, CHRNA10, ENPP1, and ADORA2A. Methodological validation showed that the established UPLC method had good linear relationship (r<0.999), and all RSD values were lower than 3% in the repeatability, stability, and precision test. Conclusion Based on the multivariate statistics, network pharmacology analysis, and UPLC multi-component quantitative analysis, a multi-angle analysis method for effective components can be established for the integral quality control of Naoxintong Capsules.
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Objective To establish an HPLC-DAD method for the simultaneous determination of amygdalin, oleanolic acid, ursolic acid, agrimol B, liquiritin, calycosin-7-glucoside, ruscogenin, gallic acid, psoralen, and shionone in Yifei Qinghua Granules (YQG). Methods The chromatographic separation was achieved on an Waters Symmetry-C18 (250 mm × 4.6 mm, 5.0 μm) column with mobile phase consisted of (1.0 g potassium phosphate monobasic in 1 000 mL water, the pH value was adjusted to 3.5 with phosphate)- (acetonitrile-methanol 1:1) for gradient elution, at the flow rate of 0.8 mL/min; The column temperature was 40 ℃. Results The linear ranges of amygdalin, oleanolic acid, ursolic acid, agrimol B, liquiritin, calycosin-7-glucoside, ruscogenin, gallic acid, psoralen, and shionone were 0.2-2.0 μg/mL (r = 0.999 4), 0.4-4.0 μg/mL (r = 0.999 4), 0.3-3.0 μg/mL (r = 0.999 6), 0.1-1.0 μg/mL (r = 0.999 3), 0.5-5.0 μg/mL (r = 0.999 1), 0.15-1.50 μg/mL (r = 0.999 2), 0.25-2.50 μg/mL (r = 0.999 5), 0.6-6.0 μg/mL (r = 0.999 1), 0.45-4.50 μg/mL (r = 0.999 3), and 0.12-1.20 μg/mL (r = 0.999 4), respectively. These ten components were well resolved. Their average recoveries (n = 6) respectively were 98.7% (RSD = 0.7%), 98.0% (RSD = 1.3%), 99.6% (RSD = 1.4%), 98.0% (RSD = 1.4%), 99.1% (RSD = 1.1%), 99.4% (RSD = 0.8%), 98.8% (RSD = 0.9%), 101.1% (RSD = 0.5%), 100.6% (RSD = 0.5%), and 101.7% (RSD = 0.8%). The content of ten batches of the amygdalin, oleanolic acid, ursolic acid, agrimol B, liquiritin, calycosin-7-glucoside, ruscogenin, gallic acid, psoralen, and shionone was 0.104-0.123, 0.600-0.621, 0.501-0.523, 0.100-0.121, 0.103-0.122, 0.231-0.260, 0.043-0.065, 0.055-0.069, 0.061-0.079, 0.031-0.043 mg/g, respectively. Conclusion The method is accurate, sensitive, credible, and repeatable, which can be applied to the quality control of YQG.
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Objective To establish a method for content determination of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province.Methods The contents of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province were determined by ultraviolet spectroscopy with calycosin-7-glucoside and glucose as reference substance, and the wavelength was set at 260 nm and 484 nm.Results The contents were from 2.82 mg/g to 6.79 mg/g for total flavones and from 106.14 mg/g to 746.40 mg/g for total polysaccharides in Hedysari Radix in different producing areas in Gansu Province. The recoveries of total flavones and total polysaccharides were 97.96％ and 102.90％, respectively.Conclusion There was difference in contents of total flavones and polysaccharides of Hedysari Radix in different producing areas in Gansu Province, and the method of using ultraviolet spectroscopy is simple, reproducible, accurate and reliable, which can be preferably used as the method for content determination of total flavones and polysaccharides in Hedysari Radix.
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Objective: To establish an HPLC-MS/MS method for the simultaneous determination of ferulic acid, tetrahydroxystilbene glucoside, puerarin, calycosin-7-glucoside, naringin, protocatechuic aldehyde, salvianolic acid B, icariin, tanshinone IIA, and synephrine in Xintong Oral Liquid (XOL). Methods: The analysis was performed on Phenomenex Luna-C18 column (250 mm × 4.6 mm, 5 μm) by gradient elution of acetonitrile-mehanol-0.02 mol/L ammonium acetate. The flow rate was 0.9 mL/min. MS conditions: electrospray ionization (ESI) and multiple reaction monitoring (MRM) were adopted, the detected peak areas of ion pairs were used for quantitative determination. Results: The linear ranges of ferulic acid, tetrahydroxystilbene glucoside, puerarin, calycosin-7-glucoside, naringin, protocatechuic aldehyde, salvianolic acid B, icariin, tanshinone IIA, and synephrine were 15.22-15 220 (r = 0.999 3), 19.52-19 520 (r = 0.999 4), 25.41-25 410 (r = 0.999 5), 35.27-35 270 (r = 0.999 2), 30.28-30 280 (r = 0.999 4), 50.11-50 110 (r = 0.999 3), 20.33-20 330 (r =0.999 2), 25.22-25 220 (r = 0.999 6), 25.36-25 360 (r = 0.999 3), and 30.29-30 290 ng/mL (r = 0.999 2). The average recoveries (n = 6) were 98.15% (RSD = 1.04%), 101.84% (RSD = 0.98%), 99.86% (RSD = 0.75%), 101.08% (RSD = 0.87%), 100.52% (RSD =1.31%), 100.67% (RSD = 1.27%), 100.46% (RSD =1.64%), 99.43% (RSD =1.48%), 100.81% (RSD = 0.67%), and 98.37% (RSD =1.16%), respectively. The contents of 10 batches of the 10 active components were 0.294-0.319, 0.640-0.665, 4.671-4.699, 0.244-0.264, 2.211-2.231, 0.180-0.201, 0.306-0.324, 1.540-1.564, 0.504-0.522, and 0.809-0.829 mg/mL. Conclusion: This method is simple and rapid, and can be used for the quality control of XOL with satisfactory separation and repeatability.
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Objective: To develop and validate an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC- MS/MS) method for the simultaneous determination of 11 components (calycosin, calycosin-7-glucoside, ginsenoside Rg1, ginsenoside Re, formononetin, astragaloside IV, ginsenoside Rf, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, and ginsenoside Rb1) in Kang'ai Injection (KI). Methods: Multiple reaction monitoring (MRM) with a polarity-switching electrospray ionization (ESI) source between positive and negative mode was used for the quantification. The 11 components were separated within 7.0 min on a Phenomenex Luna C18 column (100 mm × 3.0 mm, 2.6 μm) using a mobile phase consisted of methanol, acetonitrile, 5 mmol/L ammonium acetic-0.05 mmol/L sodium acetate water solution with gradient elution. Results: The linear relationships showed well in the area of determination. The average recovery varied between 95.2% and 104.4% with RSD ≤ 4.64%. The precision, repeatability, and stability of the method were good for the determination of 11 components. The content ranges of calycosin, calycosin-7-glucoside, ginsenoside Rg1, ginsenoside Re, formononetin, astragaloside IV, ginsenoside Rf, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, and ginsenoside Rb1 in four batches of KI were 0.004-0.006 μg/mL, 0.002-0.003 μg/mL, 125.75-148.00 μg/mL, 51.75-77.00 μg/mL, 0.010-0.013 μg/mL, 51.50-87.75 μg/mL, 27.83-30.73 μg/mL, 4.23-5.15 μg/mL, 8.40-13.35 μg/mL, 17.33-27.68 μg/mL, and 9.03-11.00 μg/mL, respectively. Conclusion: A new rapid and reliable UPLC-MS/MS method was developed and validated to evaluate the quality of KI for the first time.
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Objective: To develop a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the simultaneous determination of eight components (puerarin, tanshinone IIA, stibene glucoside, paeoniflorin, ginsenoside Re, ginsenoside Rg1, calycosin-7-glucoside, and baicalin) in Shenwu Jiannao Capsules (SJC). Methods: Chromatographic column: YMC-Triart C18, mobile phase: methanol-water (including 0.2% formic acid), and gradient elution, flow rate: 0.6 mL/min, column temperature: 40℃. MS condition: electrospray ionization (ESI) and multiple reaction monitoring (MRM) were adopted, the detected peak areas of ion pairs were used for quantitative determination. Results: There was a good linearity between the absorption peak area and the concentration for puerarin, tanshinone IIA, stibene glucoside, paeoniflorin, ginsenoside Re, ginsenoside Rg1, calycosin-7- glucoside, and baicalin in the ranges of 20.31-20 310, 25.66-25 660, 20.45-20 450, 50.79-50 790, 50.57-50 570, 50.38-50 380, 10.32-10 320, and 25.74-25 740 ng/mL, respectively (r ≥ 0.999 0). The precision was good and RSD was less than 2.0%. The repeatability was good and RSD was less than 2.0%. The stability was good in 24 h. The average recoveries were ranged from 97.98% to 102.48% (RSD ≤ 1.6%). The contents of puerarin, tanshinone IIA, stibene glucoside, paeoniflorin, ginsenoside Re, ginsenoside Rg1, calycosin-7-glucoside, and baicalin in six batches of samples were in the ranges of 1.843-1.860, 1.618-1.629, 2.116-2.129, 2.537-2.547, 0.034-0.041, 0.048-0.055, 0.551-0.564, and 2.333-2.346 mg/g, respectively. Conclusion: The method is rapid, simple, repeatable, and can be used to determine the multi components in different batches of SJC simultaneously.
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Objective:To establish an HPLC method for the determination of calycosin-7-glucoside, genistein, formononetin and medicarpin in Radix Hedysari. Methods: The sample was refluxed with methanol in a water bath. The HPLC was performed on an SunFire C18 column(250 mm × 4. 6 mm,5 μm) with acetonitrile-0. 2% H3 PO4 as the mobile phase by gradient elution. The flow rate was 1. 0 ml·min-1 and the column temperature was at 35℃. The detection wavelength was 240 nm and the sample size was 20 μl. Results:The linear range of calycosin-7-glucoside was 0.035-1.042 μg(r=0.999 6), that of genistein was 0.027-0.821 μg(r=0. 999 7), that of formononetin was 0. 031-0. 941 μg(r=0. 999 9) and that of medicarpin was 0. 025-0. 745 μg(r=0. 999 6). The average recovery of calycosin-7-glucoside, genistein, formononetin and medicarpin was 100. 32%(RSD=1. 87%), 99. 3%(RSD=1. 76%), 100. 5%(RSD=1. 48%) and 99. 2%(RSD=1. 45%)(n=6), respectively. Conclusion:The method shows short analyt-ic time, good stability and promising operation accuracy, which provides the reference for the quality control of Radix Hedysari.
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OBJECTIVE:To establish a method for simultaneous determination of the confents of calycosin-7-glucoside,calyco-sin and formononetin in Shenqi granule. METHODS:HPLC was performed on the column of Welch Ultimate XB-C18 with mobile phase of acetonitrile-0.2% Formic acid solution (gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 250 nm, column temperature was 35 ℃,and injection volume was 10 μl. RESULTS:The linear range of sample quantity was 0.008-0.32 μg for calycosin-7-glucoside(r=0.999 9),0.005-0.20 μg for calycosin(r=0.999 7)and 0.007-0.28 μg for formononetin(r=0.999 8);RSDs of precsion,stability and reproducibility tests were no more than 1.59%;recoveries were 98.17%-99.17%(RSD=0.67%, n=6),98.03%-99.66%(RSD=0.57%,n=6) and 98.16%-99.12%(RSD=0.43%,n=6). CONCLUSIONS:The method is sim-ple,accurate and reliable,and can be used for the content determination of Shenqi granule ofiso flavones.
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Objective: To establish a UPLC-MS/MS method for simultaneously determining ten components (chromogenic acid, liquiritin apioside, calycosin-7-glucoside, cynaroside, liquiritin, ferulic acid, isoliquiritoside, harpagoside, liquiritigenin, and cinnamic acid) in Tongsaimai Pellets. Methods: A method for the simultaneous determination of the ten components was established by UPLC-MS/MS. The analysis was performed on a Waters Acquity BEH C18 column (100 mm × 2.1 mm, 1.7 μm) with the mixture of acetonitrile and 0.1% formic acid/water as mobile phase, and the gradient elution at a flow rate of 0.3 mL/min. The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source combined with multiple reaction monitoring (MRM) mode. Results: The linear relationships between the concentration and peak areas of the ten components were chromogenic acid Y = 294.09 X + 17624; liquiritin apioside Y = 19.296 X + 748.42; calycosin-7-glucoside Y = 670.8 X + 11121; cynaroside Y = 490.45 X + 2938.3; liquiritin Y = 33.489 X + 555; ferulic acid Y = 127.76 X + 1343.5; isoliquiritoside Y = 57.87 X + 804.81; harpagoside Y = 13.125 X-20.365; liquiritigenin Y = 29.057 X + 367.71; cinnamic acid Y = 72.867 X + 1301.5 (r > 0.9990); The precisions, repeatabilities, and stabilities of the method were satisfactory, and the average recoveries were between 96.00% and 104.67% with the relative standard deviations no more than 3%. Conclusion: The established method is specific, rapid, and accurate for the determination of the ten effective components in Tongsaimai Pellets.
ABSTRACT
Objective: To establish a quality evaluation method of Shenqi Sherong Pill. Methods: Four types with 10 marker components were determined by HPLC. The dissolution tests were carried out with oaring method, and the accumulative release percentage was calculated. Results: The linear ranges of protocatechuic aldehyde, echinacoside, calycosin-7-glucoside, acteoside, rosmarinic acid, salvianolic acid B, calycosin, formononetin, cryptotanshinone, and tanshinone IIA were 4.14-62.1, 303.8-455.8, 1.0-150.0, 28.2-422.4, 19.0-570.0, 253.2-7596.0, 6.86-205.8, 46.4-1392.0, 54.0-1620.0, and 10.6-318.0 μg. The f2 similarity factor method indicated that the dissolution of Shenqi Sherong Pill with different lot numbers had similarity in the same media. Conclusion: The method is sensitive, stable, practical, and suitable for the quality evaluation of Shenqi Sherong Pills.