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OBJECTIVE To develop an HPLC method for the simultaneous dete rmination of morroniside ，loganin，paeoniflorin， salvianolic acid B and icariin in Shenfukang Ⅱ capsule. METHODS The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of acetonitrile- 0.1％ phosphate acid （gradient elution ）at the flow rate of 1 mL/min. The column temperature was 30 ℃，and detection wavelength was set at 240 nm. The sample size was 10 μL. RESULTS The linear range of morroniside，loganin，paeoniflorin，salvianolic acid B and icariin were 4.80-240.00，4.84-242.00，7.00-350.00，4.72-236.00 and 5.18-259.00 μg/mL（r≥0.999 8），respectively. RSDs of precision ，stability and reproducibility tests were all lower than 3％（n＝6）. Average recoveries were 97.22％-101.36％ with the RSDs of 1.19％-2.43％（n＝6）. The contents of above 5 components in 5 batches of samples were 2.019 3-2.360 0，1.624 2-1.847 1，5.637 7-6.828 0，5.015 9-5.717 0 and 1.208 8-1.754 6 mg/g，respectively. CONCLUSIONS The method is simple ，accurate and reproducible. It can improve the quality control level of Shenfukang Ⅱ capsule.
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Objective:Through comprehensive evaluation and analysis of the quality of Liuwei Dihuang （LWDH） preparations from different manufacturers and combining factors such as production technology， the key factors in the quality control of LWDH preparations are explored to provide a reference for improving the quality control level of LWDH preparations. Method:Morroniside， loganin and paeonol as quality control markers of LWDH products were determined by high performance liquid chromatography （HPLC）， the mobile phase was acetonitrile （A） -0.3% phosphoric acid aqueous solution （B） for gradient elution （0-5 min， 5%-8%A； 5-20 min， 8%A； 20-35 min， 8%-20%A； 35-45 min， 20%-60%A； 45-55 min， 60%A）， the detection wavelength of paeonol was at 274 nm， and the detection wavelengths of morroniside and loganin were at 240 nm. The quality characteristics of LWDH preparations with different dosage forms （big candied pills， water-honeyed pills， concentrated pills， hard capsules and soft capsules） from different manufacturers were analyzed. Combined these results with their actual production processes， the key-points of quality control in the whole production process were discussed. Result:The contents of three index ingredients in 128 batches of LWDH preparations were all in conformity with the standards of the 2015 edition of <italic>Chinese Pharmacopoeia</italic>， however， the content limit of some dosage forms in the current standard was unreasonable. For example， although the daily dose of crude drugs for big candied pills were almost twice the dose of water- honeyed pills （15.00， 8.57 g， respectively）， they got exactly the same daily limits of the contents for both the quality markers. What′s more， these two formulations had the same process， so the differences between the process obviously could not be the reason of these differences. Conclusion:It is recommended that for the products with different dosage forms should have a similar content limits， if there are no obvious distinctions between their production processes. Which may benefit the quality control of the products with multi-dosage forms. The research on the quality standards of proprietary Chinese medicines should deeply study the existing characteristics of the quality standards， and fully respect the laws of the quality attributes of traditional Chinese medicines and the rules of the production process of Chinese patent medicines.
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OBJECTIVE：To estab lish a me thod for simultaneous determination of morroniside ，loganin，echinacoside and acteoside in Huanshao capsules. METHODS ：HPLC method was adopted. The determination was performed on Zhongpuhong RD-C18 column with mobile phase consisted of acetonitrile- 0.1％ formic acid solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm （morroniside，loganin） and 330 nm （echinacoside，acteoside）. The column temperature was set at 35 ℃，and sample size was 10 μL. RESULTS：The linear range were 5.29-105.80 μg/mL for morroniside， 4.49-89.88 mg/L for loganin ，16.26-325.25 mg/L for echinacoside and 16.31-326.25 mg/L for acteoside ，r values were 0.999 9. RSDs of precision ，stability （24 h），reproducibility and durability tests were all lower than 2.0％ . The recoveries were 94.34％ -96.23％（RSD＝0.81％ ，n＝6），97.04％ -98.89％（RSD＝0.73％ ，n＝6），96.23％ -98.08％（RSD＝0.82％ ，n＝6）， 95.40％-98.47％（RSD＝1.23％，n＝6），respectively. The contents of above 4 components in 11 batches of Huanshao Capsules were 0.190-0.704，0.439-0.857，2.723-4.475 and 0.589-1.035 mg/g，respectively. CONCLUSIONS ：Established method is specific ， precise and can be used for content determination of 4 components in Huanshao capsules.
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Objective: To isolate and identify the terpenoids from the aerial parts of Gendarussa vulgaris. Methods: The 95% EtOH extract of the aerial parts of G. vulgaris were isolated and purified by silica gel, Sephadex LH-20, reversed-phase ODS, macroporous adsorption resin AB-8 and semi-preparative high performance liquid chromatography. The compound structures were identified by physicochemical properties and spectroscopic data. Results: Ten terpenoids were identified as gvterpennoid A (1), 4,4,14α- trimethylpregn-8-en-3β,20α-diol (2), betulone (3), ergosterol endoperoxide (4), ursolic acid (5), oleanolic acid (6), 3β-hydroxyl- 11α,12α-epoxy olean-28,13β-lactone (7), sweroside (8), loganin (9), and dehydromorroniaglycone (10). Conclusion: Compound 1 is a new triterpene, named gvterpennoid A. Compound 2 is a new natural product, and its 13C- and 1H-NMR chemical shifts were first completely assigned on the basis of 1D and 2D NMR spectroscopic evidence. Compounds 3-5, 7-10 are isolated from Gendarussa genus for the first time.
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Objective: Taking Qiju Dihuang Pills (QDP) as the research object, time domain reflection method was used for real-time determination of moisture content in concentrated pills during drying process and optimization of the drying process parameters. Methods: The moisture model of the drying process of QDP was established by the relationship between the water, temperature, and the reflective signal value of time domain reflector. The effect of the drying process on the different thickness (8, 16, and 24 mm), different drying temperatures (30, 40, 50, 60, 70, 80, and 90℃) was investigated. Results: The moisture model of the drying process of QDP was measured by time domain reflection method as Y = 0.305 X-34.772 (r2 = 0.999); X = X(T)-(0.768 9 T-24.824 7) (T ≥ 30℃). The optimized process was as following: the process was dried at 60℃ to 13.8% moisture and then rising to 80℃, after being dried to 7.80%, cooled to 60℃ and dried to 5.0% target moisture. Conclusion: It is feasible to test the moisture content in the drying process of QDP by time domain reflection method. This method can be used to monitor and popularize the moisture content in the drying process of traditional Chinese medicine concentrated pills.
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Objective: To establish a scientific and reasonable quality control method of Bufei Granules through the qualitative and quantitative research of thin layer identification and content determination of Bufei Granules. Methods: Based on the main chemical constituents of each drug in Bufei Granules, TLC method was used to analyze Corni Fructus, Ephedrae Herba, Paeoniae Radix Rubra, Scutellariae Radix, Citri Reticulatae Pericarpium, and Glycyrrhizae Radix et Rhizoma; HPLC was used to determine the content of loganin and baicalin. The content of loganin was analyzed by Agilent Eclipse XDB-C18 column (250 mm × 4.6 mm, 5 μm) with the mobile phase of acetonitrile-methanol-water-formic acid (10:1:89:0.1). The detection wavelength was set at 236 nm; The content of baicalin was analyzed by Dikma Diamonsil C18 column (250 mm × 4.6 mm, 5 μm) with the mobile phase of methanol-water-formic acid (49:51:0.1). The detection wavelength was set at 280 nm. Results: The TLC identification method can distinguish Ephedrae Herba, Paeoniae Radix Rubra, Scutellariae Radix, Citri Reticulatae Pericarpium, and Glycyrrhizae Radix et Rhizoma with clear spots, no negative interference, good separation and strong specificity. Loganin and baicalin were used as index components in methodological study. The average recovery of loganin was 98.49% and the RSD was 0.80%; The repeatability test RSD was 0.83% which met the requirement. The linear range was from 4.76 μg/mL to 50.70 μg/mL and the linear relationship was good (r = 0.999 9). The average recovery of baicalinrate was 101.20% and the RSD was 0.77%. The repeatability test RSD was 0.90%, which met the requirements. The linear range was from 6.00 μg/mL to 96.00 μg/mL and the linear relationship was good (r = 0.999 9). The loganin content of three bantches of samples was 12.04, 9.78 and 11.81 mg/bag; And the content determination result of baicalin was 121.13, 101.31 and 103.14 mg/bag. Conclusion: The method is easy to operate, strong in specificity, accurate and sensitive, with good repeatability. It can be applied for the quality control of Bufei Granules.
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OBJECTIVE：To study the effects of loganin on the prolife ration and apoptosis of liver cancer HepG 2 cells，and to explore its mechanism. METHODS ：CCK-8 assay was used to detect the effects of different concentrations （10，25，50，100， 150，200，300，400 µg/mL）of loganin on the proliferation activity of HepG 2 cells for 24 and 48 h. HepG 2 cells were divided into control group ，loganin low-concentration ，medium-concentration and high-concentration groups （50，100，150 μ g/mL）. After treated for 24 h，morphological changes of apoptosis of cells were detected by Hoechst 33342 fluorescence staining. The apoptosis and cycle distribution of cells were detected by flow cytometry. Western blotting was used to detect protein expression of Cyclin D1， PCNA， Bcl-2， Caspase-3， Cleaved-Caspase-3， Caspase-9 and Cleaved-Caspase- 9. RESULTS ： Loganin inhibited the proliferation of HepG 2 cells，in concentration-dependent trend. Compared with control group ，apoptosis as pyknosis and fragmentation occurred ，and the apoptosis rate increased significantly in loganin low-concentration ，medium-concentration and high-concentration groups （P＜0.01）；the cell were mainly blocked in S phase ；relative protein expression of Cyclin D 1，PCNA and Caspase- 3 were significantly decreased ，while that of Cleaved-Caspase- 3 were significantly increased in loganin low- concentration， medium-concentration and high-concentration groups （P＜0.05 or P＜0.01）； relative protein expression of Cleaved-Caspase-9 were increased significantly ，while that of Bcl- 2 and Caspase- 9 were decreased significantly in loganin medium-concentration and high-concentration groups （P＜0.05 or P＜0.01）. CONCLUSIONS ：Loganin can significantly inhibit the proliferation and induce apoptosis of HepG 2 cells，the mechanism of which may be associated with inhibiting Bcl- 2 protein expression and promoting Caspase- 3，Caspase-9 activation.
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Objective: To compare the differences in pharmacokinetic behavior of six ingredients in Qikui Sustained-release Tablets in rabbit plasma. Qikui Granules was taken as reference. Methods: Diazepam was used as internal standard. LC-MS/MS detection methods of astragaloside, hyperin, isoquercitrin, rutin, morroniside, and loganin in rabbit plasma were established, and pharmacokinetic parameters of six components were calculated. Results: Six active ingredients’ equation of linear regressions were: astragaloside Y = 1.0 × 10-4 X - 0.009 9 (r = 0.999 7), morroniside Y = 1.0 × 10-4 X + 0.038 7 (r = 0.999 4), loganin Y = 3.0 × 10-5 X + 0.008 7 (r = 0.999 3), hyperin Y = 1.0 × 10-3 X - 0.016 1 (r = 0.999 0), rutin Y = 5.0 × 10-4 X - 0.011 5 (r = 0.999 4), isoquercitrin Y = 1.7 × 10-3X - 0.307 5(r = 0.999 2). Intra-day and inter-day precision and accuracy and recovery rate were up to the mustard. After Qikui Sustained-release Tablets and Qikui Granules being given by gavege, the maximal concentration (Cmax) of morroniside, loganin, astragaloside, rutin, hyperin, and isoquerctirin in Qikui Granules were (1.333 ± 0.051), (1.238 ± 0.164), (0.83 ± 0.079), (0.127 ± 0.017),(0.444 ± 0.048), and (0.223 ± 0.048) mg/L, t1/2 were (3.848 ± 0.311), (3.822 ± 0.757), (4.982 ± 1.14), (3.73 ± 0.298), (4.732 ± 0.642), and (5.132 ± 0.901) h, respectively, AUC(0-t) were (3.069 ± 0.307), (2.891 ± 0.943), (2.079 ± 0.306), (0.313 ± 0.068), (1.087 ± 0.177), (0.496 ± 0.129) mg∙h/L, respectively, Cmax of morroniside, loganin, astragaloside, rutin, hyperin, and isoquerctirin in Qikui Sustained-release Tablets were (0.985 ± 0.13), (0.961 ± 0.175), (0.693 ± 0.101), (0.094 ± 0.012), (0.354 ± 0.045), (0.201 ± 0.037) mg/L, t1/2 were (4.691 ± 0.337), (5.62 ± 1.64), (6.408 ± 0.707), (4.103 ± 0.341), (6.048 ± 0.882), (5.803 ± 0.59) h, AUC(0-t) were (5.191 ± 1.046), (6.168 ± 1.25), (4.293 ± 0.823), (0.485 ± 0.103), (1.84 ± 0.432), (0.924 ± 0.19) mg∙h/L. Contrast with Qikui Granules, relative bioavailability of morroniside, loganin, astragaloside, rutin, hyperin, and isoquerctirin in Qikui Sustained-release Tablets were 169.1%, 213.3%, 206.5%, 156.0%, 169.3%, and 186.3%, respectively. Conclusion: Qikui Sustained-release Tablets can significantly improve the bioavailability of each active ingredient in rabbit.
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OBJECTIVE： To establish the method for the rapidly non-destructive quality control of Liuwei dihuang capsule. METHODS： AOTF-NIR spectrometry was adopted. Taking 80 batches of Liuwei dihuang capsule produced by a manufacturer in recent three years as samples， HPLC chromatogram was adopted to determine the contents of loganin， morroniside， paeonol， paeoniflorin and ursolic acid； the content of water was determined according to general principles stated in 2015 edition of Chinese Pharmacopeia （part Ⅰ）. Taking 70 batches of samples as correction set， the partial least square method and the cross-validation algorithm were used to establish the NIR quantitative model of 6 indexes in Liuwei dihuang capsules with the Unscrambler quantitative analysis software. Taking residual 10 batches of samples as validation set， external validation was conducted for the model. RESULTS： The correlation coefficients （R2） of internal and external validation of loganin， morroniside， paeonol， paeoniflorin， the content of water quantitative model were all greater than 0.9； the correction of standand deviation （RMSEC） were 0.372 8， 0.025 4， 0.263 3， 0.288 5， 0.186 7 and 0.037 7； the prediction of standard deviation （RMSEP） were 0.462 2， 0.077 5， 0.472 1， 0.634 9， 0.293 4 and 0.206 9； the external verification showed that mean deviations of preclicted value to actual value were 6.04％， 6.05％， 5.87％， 6.97％， 5.62％ and 4.83％， with the mean deviation less than 10％.CONCLUSIONS：The established method can achieve rapidly non-destructive analysis Liuwei dihuang capsule.
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AIM To establish an HPLC method for the simultaneous content determination of chlorogenic acid,loganin,paeoniflorin,baicalin,salvianolic acid B and paeonol in Tuiyin Mixture (Moutan Cortex,Paeoniae Radix Rubra,Lonicerae japonicae Caulis,etc.).METHODS The analysis of methanol extract of this drug was performed on a 30 ℃ thermostatic ZORBAX SB-Aq column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 235 nm.RESULTS Six constituents showed good linear relationships within their own ranges (r≥0.999 7),whose average recoveries were 97.1％-100.6％ with the RSDs of 1.35％-2.28％.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Tuiyin Mixture.
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AIM To establish an HPLC-DAD method for the simultaneous content determination of six constituents in Maiwei Dihuang Pills (Ophiopogonis Radix,Schisandrae chinensis Fructus,Rehmanniae Radix Praeparata,etc.).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Agilent ZORBAX SB-C18column (4.6 mm × 150 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.2％ phosphate acid) flowing at 0.8 mL/min in a gradient elution manner,and the detection wavelengths were set at 220,230,236 and 274 nm.RESULTS Deoxyschizandrin,schizandrin B,schisandrin,paeoniflorin,paeonol and loganin showed good linear relationships within the ranges of 10-70,6.5-45.5,33.5-234.5,17-119,31-217 and 34-238 μg/mL (r ＞0.990 0),whose average recoveries (RSDs) were 99.6％ (1.7％),100.4％ (1.8％),100.7％ (1.8％),102.9％ (1.7％),102.2％ (1.5％) and 99.7％ (1.2％),respectively.CONCLUSION This simple and reproducible method can be used for the rapid quality control of Maiwei Dihuang Pills.
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Objective To establish an HPLC method for the content determination of morroniside, sweroside, paeoniflorin and loganin of Liuwei Dihuang Decoction and its Cornus Officinalis-Cortex Moutan couple; To discuss the relationship between the whole prescription and the couple of main pharmacodynamic components. Methods The HPLC method was used at Hypersile C18 column (4.6 mm × 250 mm, 5 μm); the mobile phase consisted of methanol-water (24:76); the detective wavelength was 236 nm; the flow rate was 1.0 mL/min; the column temperature was 30 ℃. Results The linear ranges of morroniside, sweroside, paeoniflorin and loganin were among 0.480–7.680 μg (r=0.999 3), 0.103–1.650 μg (r=0.999 5), 0.120–1.920 μg (r=0.999 1) and 0.227–3.630 μg (r=0.999 7), respectively. The average recovery rates and RSD were 102.79%, 102.29%, 100.99%, 102.48%, and 1.73%, 1.48%, 1.32%, 0.75%, respectively. The contents of morroniside, sweroside and paeoniflorin in Liuwei Dihuang Decoction were slightly higher than that in Cornus Officinalis - Cortex Moutan couple, and the contents of loganin were almost the same. Conclusion The method is simple, stable, accurate and reproducible. It can be used for content determinate of glycosides in Liuwei Dihuang Decoction and Cornus Officinalis-Cortex Moutan couple. Cornus Officinalis-Cortex Moutan couple has the glycosides with tonifying kidney effect of Liuwei Dihuang Decoction.
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To establish the HPLC fingerprint and determine five index components (loganic acid， chlorogenic acid， loganin， sweroside and asperosaponin Ⅵ) of Zishen Yutai pills by high performance liquid chromatography, and provide a scientific basis for its quality control. The fingerprint chromatogram was analysed by the chromatographic fingerprint similarity evaluation system for tradition Chinese medicine (2012), fifteen common peaks were obtained at the wavelength of 254 nm. Different batches of Zishen Yutai pills showed a similarity of above 0.90 in HPLC fingerprint profiles. For the quantitive analysis method, The separation of five components showed good regression (>0.999 2) with linear ranges, and the mean recoveries were in the range of 97.62%-101.9%, with the RSD (=9) less than 3%. The established fingerprint and quantitative analysis methods are highly specific, simple and accurate, which can reflect the quality of Zishen Yutai pills more comprehensively, and can be used for its quality control.
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Objective To study the chemical constituents from the leaf of Syringa oblata. Methods The compounds was isolated by silica gel column chromatography and HPLC, and their structure were identified by spectral data analysis. Results A total of 35 compounds were isolated and identified as oleanolic acid (1), ursolic acid (2), betulinic acid (3), 1,3-benzodioxole-5-propanol (4), p-hydroxyl benzene propyl alcohol (5), p-hydroxyl benzene ethel alcohol (6), syringopicrogenin D (7), syringopicrogenin E (8), syringopicrogenin F (9), syringopicrogenin A (10), syringopicrogenin C (11), 3,4-dihydroxyl benzene ethel alcohol (12), syringobittergenin B (13), syringo-picrogenin B (14), grasshopper ketone (15), (7R,8S)-4,9,9’-trihydroxyl-3,3’-dimethoxyl-7,8- dihydrobenzofuran-1’-propylneolignan (16), lariciresinol (17), syringin (18), 3(Z)-enol glucoside (19), quercetin-3-O-β-D- glucopyranoside (20), (8E)-ligstroside (21), epipinoresinol-4-O-β-D-glucopyranoside (22), (8E)-ligstroside B (23), (8E)-ligstroside A (24), salidroside (25), 7-dehydrologanin (26), fliederoside B (27), syringopicroside B (28), oleoside dimethyl ester (29), lilacoside (30), syrigopicroside (31), oleuropein (32), (+)-lariciresinol-4-O-β-D-glucopyranoside (33), verbascoside (34), and (+)- epipinoresinol-4’-O-β-D-glucopyranoside (35). Conclusion Compounds 4, 5, 14-16, 19, 23, 24, 26, and 27 are isolated from S. oblata for the first time.
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AIM To establish a quantitative analysis of multi-components by single mark (QAMS) method for the simultaneous content determination of five constituents in Shanzhuyu Formulated Granules (Corni Fructus).METHODS The analysis of 80％ methanol extract of this drug was performed on a 30 ℃ thermostatic Wondasil C18column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.3％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 335 nm.With morroniside as an internal standard,the relative correction factors of gallic acid,5-hydroxymethylfurfural,loganin and cornuside were established,followed by the determination of their contents.RESULTS Gallic acid,5-hydroxymethylfurfural,morroniside,loganin and cornuside showed good linear relationships within the ranges of 0.0120-0.120,0.026 8-0.268,0.074 4-0.744,0.058 6-0.586 and 0.0086-0.086 μg (r≥ 0.9999),whose average recoveries (RSDs) were 103.43％ (1.45％),103.36％ (1.50％),104.47％ (0.30％),102.08％(1.74％) and 104.01％ (0.62％),respectively.The results obtained by QAMS approximated those obtained by external standard method.CONCLUSION This stable and reliable method can be used for the quality control of Shanzhuyu Formulated Granules.
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Objective To establish the quality control methods for Qihuang Fuzheng particle .Methods Astragali Ra-dix ,Rehmanniae Radix Praeparata ,Corni Fructus ,Chuanxiong Rhizoma ,Angelicae Sinensis Radix , Paeoniae Radix Rubra ,Moutan Cortex ,and Ophiopogonis Radix were identified with TLC .HPLC method was used for loganin assay in Cor-ni Fructus .Results TLC identification methods for the main components were established .The TLC spots were clear with good separation .No interference was detected from the negative samples .The peak response and concentration of loganin showed good linear relationship over the range of 4 .02-80 .40 μg/ml (r=0 .9999) .The mean recovery of loganin was 99 .02％(RSD=0 .64％ ,n=9) .Conclusion The established quality control methods are accurate ,reliable ,and specific ,which lay a foundation for the quality control of Qihuang Fuzheng particle .
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AIM To evaluate the stability of Zuogui Concentrated Pills.METHODS HPLC was applied to determining the contents of acteoside,loganin and hyperoside,whose relative contents were detected by classical constant temperature accelerated test,after which the fitting of kinetic equation was conducted.The period of validity was predicted by classical constant temperature method and multivariate linear model,and the stability was investigated by high temperature,high humidity,strong light,accelerated and long-term tests.RESULTS The degradation of acteoside,loganin and hyperoside accorded with the first-order kinetic process.The periods of validity were found to be 25.6 months and 23.9 months by two methods,respectively.No obvious changes were observed on three constituents' contents,appearance and character,disintegration time limit,moisture content and weight variation under various tests.CONCLUSION A tentatively scheduled two-year validity period is suitable for Zuogui Concentrated Pills due to its good stability.
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Objective: To establish methods of qualitative identification and quantitative determination for Zishen Yangyin Granules (ZYG). Methods: The TLC method was used to identify the herb by mixture of chloroform-methanol (31) as a developing solvent on high performance silica gel precoated plate (HSGF254) and using 5% vanillic aldehyde sulfuric acid as a chromogenic reagent for qualitative identification of Corni Fructus; TLC identification of Eclipta prostrata, Alismatis Rhizoma, and Moutan Cortex was performed on high performance silica gel precoated plate (HSGF254) with petroleum ether (60-90 ℃)-chloroform-ethyl acetate (312) as developing solvent. The same developing method was used to identify E. prostrata, A. Rhizoma, and paneol in M. Cortex of ZYG by different detected method at the same time. The contents of morroniside, loganin, hyperoside, specnuezhenide, and paeonol were analyzed by high performance liquid chromatography on C18 column (250 mm × 4.6 mm, 5 μm) with the mobile phase of acetonitrile-0.1% trifluoroacetic acid by gradient elution. The detection wavelength was set at 254 nm. Results: Morroniside and loganin were used to identify C. Fructus in ZYG, paeonol in M. Cortex can be identified at 254 nm; Some substances in E. prostrata can be identified at 366 nm; Some substances in A. Rhizoma can be detected in sunlight, with 5% phosphomolybdic acid in ethanol as a chromogenic reagent. The TLC separation was desirable with moderate Rf value and clear spot. The methodology validation for the assay of morroniside, loganin, hyperoside, specnuezhenide, and paeonol presented that they were in good linear correlation in the ranges of 4.432-110.8, 4.192-104.8, 4.040-101.0, 4.132-103.3, and 4.076-101.9 μg/mL, The correlation coefficients of indicator were over 0.999 7. The average recoveries were between 96.57% and 98.67%. The RSD value of intra-day precision was less than 2% and the RSD value of inter-day precision was less than 3%. The method has good stability and reproducibility. Conclusion: The methods of quality control are specific, reproducible, accurate, and suitable, which can be successfully applied to the quality control of ZYG.
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AIM To establish a quantitative analysis of multi-components by single mark (QAMS) method for the simultaneous content determination of five constituents in Shanzhuyu Formulated Granules (Corni Fructus).METHODS The analysis of 80％ methanol extract of this drug was performed on a 30 ℃ thermostatic Wondasil C18column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.3％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 335 nm.With morroniside as an internal standard,the relative correction factors of gallic acid,5-hydroxymethylfurfural,loganin and cornuside were established,followed by the determination of their contents.RESULTS Gallic acid,5-hydroxymethylfurfural,morroniside,loganin and cornuside showed good linear relationships within the ranges of 0.0120-0.120,0.026 8-0.268,0.074 4-0.744,0.058 6-0.586 and 0.0086-0.086 μg (r≥ 0.9999),whose average recoveries (RSDs) were 103.43％ (1.45％),103.36％ (1.50％),104.47％ (0.30％),102.08％(1.74％) and 104.01％ (0.62％),respectively.The results obtained by QAMS approximated those obtained by external standard method.CONCLUSION This stable and reliable method can be used for the quality control of Shanzhuyu Formulated Granules.
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Objective To establish an HPLC fingerprint of Xiaochuan Granula (XCG), and to make a quantitative analysis of seven components by fused-core column. Methods Kromasil C18 (150 mm × 4.6 mm, 3.5 μm) was used with the mobile phase of Methanol (A)-acetonitrile (B)-water (C), at a flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm for amygdalin and magnolin, 240 nm for morroniside, loganin, prim-O-glucosylcimifugin, 4'-O-β-glucopyranosyl-5-O-methylvisamminol, schizandrin, and fingerprint; And the column temperature was maintained at 40 ℃. Common peaks had been identified by UPLC-Q-TOF/MS and standard compounds. Results The fingerprint chromatography included 20 mutual peaks, and the similarity was more than 0.90. Eleven chemical components were identified by UPLC-Q-TOF/MS and standard compounds, which were 3-morroniside, oxypaeoniflorin, 5-loganin, 6-prim-O-glucosylcimifugin, 9-4'-O-β-glucopyranosyl-5-O-methylvisamminol, 11-magnolin, 15-schisandrin, 16-schisandrol B, 18-schisantherin A, 19-deoxyschizandrin, and 20-γ-schizandrin B. Moreover, seven active components (morroniside, oxypaeoniflorin, loganin, prim-O-glucosylcimifugin, 4'-O-β-glucopyranosyl-5-O-methylvisamminol,magnolin, and schisandrin) were quantified and the average recovery rates ranged from 97.3% to 103.8% with RSDs less than 2.0%; Seven components in 10 batches samples were morroniside 0.51-0.69 mg/g, amygdalin 5.01-5.95 mg/g, loganin 1.02-1.33 mg/g, prim-O-glucosylcimifugin 0.35-0.45 mg/g, 4'-O-β-glucopyranosyl-5-O-methylvisamminol 0.45-0.55 mg/g, magnolin 0.38-0.48 mg/g, schisandrin 0.89-1.08 mg/g, respectively, and RSD of each component was less than 11.0%. Conclusion The method for establishing HPLC fingerprint and quantitative analysis of seven components is rapid, simple, and accurate, and can be used for the quality control of XCG.