Study on scrophulariaceae radix reference extract and its application in quality control of Scrophulariae Radix / 中国中药杂志

By establishing the preparation process of Scrophulariaceae Radix reference extract(SRRE) and calibrating it, we discussed its feasibility as a substitute for single reference substance in the quality control of Scrophulariae Radix. The SRREs were prepared by solvent extraction method and chromatographic separation technology, and then calibrated with the reference substances of harpagide, angoroside C and harpagoside. The HPLC content determination method of Scrophulariae Radixl was established with SRREs of the known content and the reference substances of harpagide, angoroside C and harpagoside respectively as the control ones. Then the content of three components in Scrophulariae Radix was determined, and the t-test method was used to compare the results of the two methods. With SRRE as references, harpagide, angoroside C and harpagoside were in a good linear relationship(r≥0.999 8) within each range, and the average recovery rate was 98.55% to 100.6%. The t-test results showed that the P values of two determination methods were 0.493, 0.155 and 0.171 for harpagide, angoroside C and harpagoside respectively, indicating no significant diffe-rence between the two methods of content determination. The SRRE can be used as a substitute for the reference in the quality control of Scrophulariaceae Radix. The SRRE can replace the corresponding reference substance for the quality control of Scrophulariae Radix. The results of this study provide new methods and new ideas for the quality evaluation of Scrophulariae Radix, and provide a scientific basis for the application of reference extracts in the quality research of traditional Chinese medicine.

Determination of harpagide and harpagoside contents in Scrophularia ningpoensis from 15 origins / 药学实践杂志

Objective To establish an HPLC method for the simultaneous determination of harpagide and harpagoside content in Scrophularia ningpoensis (SN). Methods An Eclipse C18 column was used for determination of methanol extract of S. ningpoensis with a HPLC-PDA method and mobile phase of acetonitrile-0.03% phosphate solution in a gradient elution manner. The flow rate of mobile phase was 1.0 ml/min, and the detection wavelengths were 210 nm and 280 nm. Results Harpagide and harpagoside contents in SN showed good linear relationships within 0.1020-0.5100 mg/ml (r=0.9999) and 0.0340-0.1700 mg/ml (r=0.9999). Their average recovery rates were 97.44% and 97.08%.The RSDs were 0.93% and 1.24%.. There were significant differences in the contents of harpagide and harpagoside in SN from 15 origins (P<0.01). The content of harpagoside in Sichuan Long-dong, Zhejiang Lin-an, Zhejiang Pan-an and Henan Nan-feng is higher. Conclusion This method is stable, accurate and reproducible and can be used for the quality control of SN.

Isolation and identification of endophytic fungi producing harpagoside and harpagide from Scrophularia ningpoensis / 中国中药杂志

The endophytic fungi from root,main stem,branch and leaf of Scrophularia ningpoensis were isolated from Zhejiang,whether these strains could yield harpagide or harpagoside were tested by HPLC and LC-MS. According to the morphological characteristic and the similarity of the nucleotide sequence of internal transcribed spacer( ITS) between r DNAs,the strains producing harpagide or harpagoside were identified. The results showed that 210 strains were isolated from the samples,which were classified into 9 orders,13 families and 17 genera by morphological study. Harpagide was detected in endogenous fungi ZJ17 and harpagoside was detected in endogenous fungi ZJ25 by HPLC coupled with LC-MS. ZJ17 was identified as Alternaria alternate and ZJ25 was identified as A.gaisen by its morphology and authenticated by ITS( ITS4 and ITS5 regions and the intervening 5. 8 S rDNA region).

Grade quality of Scrophulariae Radix based on fingerprints and quantitative analysis of multi-index / 中草药

Objective In order to provide a scientific basis for the grade quality standard criterion of Scrophulariae Radix, a HPLC-DAD method was established to acquire fingerprint and detect the content of multiple components. Methods The chromatographic separate was achieved on Elipse XDB-C18 (250 mm × 4.6 mm, 5 μm) column, with the temperature of 25 ℃, using acetonitrile (A)-0.03% phosphoric acid water (B) as the mobile phase gradient elution, a flow rate of 1.0 mL/min, and the detection wavelength was set at 207 nm for fingerprint, 210 nm for Harpagide, 280 nm for harpagoside, and 264 nm for cinnamic acid. The injection volume was 10 μL. The fingerprints of different grades of Scrophulariae Radix from 30 batches were evaluated with a chromatographic fingerprint similarity evaluation system (version 2012), and the content of harpagide, harpagoside, and cinnamic acid was also determined at the same time. Results There were seven common peaks in the fingerprints, the RSD values of relative retention time were lower than 2.0% and those of relative peak area were quite different, which indicated that the main chemical compounds can exist stably in Scrophulariae Radix with different content. Compared with control, the results of fingerprintsimilarity were presented as follows: 5% under 0.7, 12.5% arranged 0.7 to 0.8, 40% between 0.8 and 0.9, and another 42.5% exceed 0.9, demonstrated that there was qualitative difference in various batches of Scrophulariae Radix. Also, the quantitative analysis of multi-index showed the differences in three main compounds, the content respectively was 0.18%—2.89% in harpagide, 0.01%—0.35% in harpagoside, and 0.01%—0.24% in cinnamic acid. Conclusion Suggestions were provided in the formulation of new grade quality standard, such as adding the fingerprints and multi-index detection of main chemical components based on the original criterion of classification.

Optimization of steaming technology for decoction pieces of Scrophularia ningpoensis by D-optimal response surface methodology based on AHP- entropy weight method / 中草药

Objective :To determine the weight coefficient and optimize the steaming technology of decoction pieces of Scrophularia ningpoensis (SN). Methods The contents of harpagide, harpagoside, aucubin, acteoside, angoroside C, and cinnamic acid were simultaneously determined by HPLC. The weight coefficient of each component was evaluated by AHP-entropy (analyitc hierarchy process) method. With composite score as index, D-optimal response surface methodology was adopted to investigate the effects of soaking time, steaming time, and drying temperature on the quality of processed products and optimize the processing technology of decoction pieces of SN. Results :Optimal processing parameters were as follows: soaking time was 15.63 min, steaming time was 85 min, drying temperature was 60 ℃, and the synthetical mark was 97.20. Considering the actual situation, the optimum processing technology of SN was obtained by fine-turning the soaking time. The soaking time was 15 min, the steaming time was 85 min, and the drying temperature was 60 ℃. Also, the synthetical mark were 98.53, 99.39, 98.86, and its RSD was 0.47% through the obtained conditions in parallel with three batches of samples. Conclusion: The optimized steaming technology is simple and feasible, which can provide a reference for the steaming of decoction pieces of SN. The method established to simultaneously determine the contents of six components in decoction pieces of SN is rapid and reliable for controlling the quality of decoction pieces of SN.

Binding interaction of harpagoside and bovine serum albumin: spectroscopic methodologies and molecular docking / 中国中药杂志

Scrophularia ningpoensis has exhibited a variety of biological activities and been used as a pharmaceutical product for the treatment of inflammatory ailment, rheumatoid arthritis, osteoarthritis and so on. Harpagoside (HAR) is considerer as a main bioactive compound in this plant. Serum albumin has important physiological roles in transportation, distribution and metabolism of many endogenous and exogenous substances in body. It is of great significance to study the interaction mechanism between HAR and bovine serum albumin (BSA). The mechanism of interaction between HAR and BSA was investigated using 2D and 3D fluorescence, synchronous florescence, ultraviolet spectroscopy and molecular docking. According to the analysis of fluorescence spectra, HAR could strongly quench the fluorescence of BSA, and the static quenching process indicated that the decrease in the quenching constant was observed with the increase in temperature. The magnitude of binding constants (KA) was more than 1×10⁵ L·mol⁻¹, and the number of binding sites(n) was approximate to 1. The thermodynamic parameters were calculated through analysis of fluorescence data with Stern-Volmer and Van't Hoff equation. The calculated enthalpy change (ΔH) and entropy change (ΔS) implied that the main interaction forces of HAR with BSA were the bonding interaction between van der Waals forces and hydrogen. The negative values of energy (ΔG) demonstrated that the binding of HAR with BSA was a spontaneous and exothermic process. The binding distance(r) between HAR and BSA was calculated to be about 2.80 nm based on the theory of Frster's non-radiation energy transfer, which indicated that energy is likely to be transfer from BSA to HAR. Both synchronous and 3D florescence spectroscopy clearly revealed that the microenvironment and conformation of BSA changed during the binding interaction between HAR and BSA. The molecular docking analysis revealed HAR is more inclined to BSA and human serum albumin (HSA) in subdomain ⅡA (Sudlow's site I). This study will provide valuable information for understanding the action mechanism of HAR.

Simultaneous determination of six constituents in Xuanmai Ganju Granules by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of six constituents in Xuanmai Ganju Granules (Scrophulariae Radix,Ophiopogonis Radix,Glycyrrhizae Radix et Rhizoma,Platycodonis Radix).METHODS The analysis of 80％ methanol extract of this drug was performed on a 35 ℃ thermostatic ZORBAX SB-C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 210,250,278 nm.RESULTS Harpagide,liquiritin apioside,liquiritin,harpagoside,cinnamic acid and glycyrrhizic acid showed good linear relationships within the ranges of 2.177-43.539 μg/mL(r =0.999 6),1.713-34.261 μg/mL (r =0.999 5),1.946-38.916 μg/mL(r =0.999 6),2.070-41.395 μg/mL(r =0.999 7),2.06-41.2 pg/mL (r =0.999 6) and 3.623-72.454 μg/mL (r =0.999 6),whose average recoveries (RS-Ds) were96.08％ (2.1％),95.55％ (2.5％),95.04％ (2.6％),94.86％ (2.7％),95.70％ (1.9％) and 95.47％ (1.9％),respectively.CONCLUSION This simple and accurate method can be used for the quality control of Xuanmai Ganju Granules.

Establishment of the Quality Standard for Xiaotang Zhike Pills / 中国药师

Objective:To establish the quality standard for Xiaotang Zhike pills.Methods:Atractylodis Rhizoma,Puerariae Lobatae Radix and Scrophulariae Radix in the formula were qualitatively identified by TLC. HPLC was used for the determination of puerarin in Xiaotang Zhike pills. The analysis was performed on a Wondasil C18-WR(250mm × 4.6 mm,5 μm)column with methanol-water(25:75) as the mobile phase at a flow rate of 1.0 ml·min-1. The detection wavelength was set at 250 nm, the column temperature was 40℃ and the sample size was 10 μl. Results:TLC showed good specificity without any interference for the identification of atrctylodin,puerarin and harpagoside. The calibration curve of puerarin was in good linearity over the range of 41.20-618.00 ng(r = 0.999 9). The average recovery was 100.16%(RSD = 2.91%). Conclusion:The established methods in the study are simple,feasible and reproducible,which are suitable for the quality control of Xiaotang Zhike pills.

Chemical constituents from Simiao Yongan Decoction / 中草药

Objective: To study the chemical constituents of Simiao Yongan Decoction. Methods: The silica gel, ODS, Sephadex LH-20, and AB-8 macroporous adsorption resin column chromatography were used to isolate and purify the chemical constituents from Simiao Yongan Decoction. The structures of the constituents were identified on the basis of physiochemical properties, NMR, MS, and other data. Results: Twenty-two compounds were isolated and identified as 5(S)-5-carboxystrictosidine (1), harpagoside (2), geniposide (3), glycyrrhetinic acid (4), glycyrrhizic acid (5), hyperoside (6), liquiritin (7), isoliquiritoside (8), liquiritigenin (9), isoliquiritigenin (10), luteolin (11), quercetin (12), 2-(3-hydroxy-4-methoxyphenyl)ethyl O-α-arabinopyranosyl-(1→6)-O-α- rhamnopyranosyl-(1→3)-O-β-glucopyranoside (13), angroside C (14), acteoside (15), cinnamic acid (16), ferulic acid (17), (E)-aldosecologanin (18), protocatechuic acid (19), stigmasterol (20), hentriacontanol (21), and daucosterol (22). Conclusion: Compounds 1-3, 5, 6, 8, 11, 13-15, 18-21 are isolated from Simiao Yongan Decoction for the first time.

Application of QAMS in quality control of Scrophularia Radix / 中草药

Objective To establish a measurement method for the content of five compounds (acteoside, harpagide, harpagoside, angoroside-C, and cinnamic acid) in Scrophularia Radix quantitative analysis multi-components by single-marker (QAMS), and verify the accuracy and feasibility of QAMS in the quality control. Methods Taking harpagide as internal standard substance, the relative correlation factor (RCF) of acteoside, cinnamic acid, harpagoside, and angoroside C was established. And the content of each component in Scrophularia Radix was determined by the above-mentioned RCF. In order to prove the scientificity and feasibility of this method, the results were compared with the external standard method. Results The relative correction factors of acteoside, cinnamic acid, harpagoside, and angoroside-C were 0.068 (RSD = 0.53%), 0.060 (RSD = 0.81%), 0.142 (RSD = 1.17%), 0.197 (RSD = 1.82%). No significant differences were found among the quantitative results of four components in 25 batches of Scrophularia Radix determined by the two methods. Conclusion It is feasible and accurate to evaluate the quality of Scrophularia Radix by QAMS.

Optimization of Extraction Technology of Panax Notoginseng and Scrophulariae Radix from Rupixiao Granule by Orthogonal Design / 中国药师

Objective: To optimize the extraction technology of Panax notoginseng and Scrophulariae radix from Rupixiao granule.Methods: With the dry extract rate and transfer rates of ginsenoside Rg1, ginsenoside Rb1 and harpagoside as the comprehensive index, the orthogonal design was adopted to investigate the effects of the amount and concentration of ethanol, extracting duration and times on the extraction technology.The contents of ginsenoside Rg1, ginsenoside Rb1 and harpagoside were determined by HPLC.Results: The optimal extraction technology was extracted twice with 8-fold amount of 60% ethanol with 2 h per time.The transfer rate of ginsenoside Rg1, ginsenoside Rb1 and harpagoside was (79.4%±1.56%), (42.62%±0.68%) and (44.89%±0.58%)(n=3), respectively.The dry extract rate was (20.99%±0.411%).Conclusion: The optimized extraction technology is stable and feasible, which can be used for extracting Panax notoginseng and Scrophulariae radix from Rupixiao granule.

Spectroscopic Study of Interaction of Harpagoside and Human Serum Albumin / 分析化学

Harpagoside (HAR) is believed to be a main compound in Scrophularia ningpoensis which possess a broad of biological activities.Human serum albumin (HSA) has important physiological roles in transportation, distribution and metabolism of many endogenous and exogenous substances in body.It is great significance in pharmacology to investigate the interaction mechanism of HAR and HSA.In this work, the interaction between HAR and HSA was investigated by fluorescence and ultraviolet absorption spectroscopy at different pH (pH=4.0, 7.4, and 9.0) and temperatures (297, 310 and 323 K).The experimental results showed that the HAR could cause the fluorescence quenching of HSA through a static quenching procedure, showing that the HAR regularly quenched the intrinsic fluorescence of HSA, and a decrease in the quenching constant was observed with an increase in temperature.Under different conditions, all the magnitude of binding constants (KA) was larger than 105 L/mol and the number of binding sites (n) in the binary system were approximate to 1.Base on the magnitude of enthalpy and entropy changes, the negative values of ΔG, ΔH and ΔS revealed that the binding of HAR with HSA was spontaneous and exothermic process, and the main interaction forces of the HAR with HAR were van der Waals forces and/or hydrogen bonding interaction.The binding distance (r) between the HAR and HSA was calculated to be about 4.2 nm based on the theory of F(o)rster′s nonradiation energy transfer, which indicated that the energy transfer from HSA to HAR occurred with high possibility.What was more, the synchronous florescence spectroscopy confirmed the conformational changes of HSA during the binding reaction.

Quality assessment of Yinju Jiedu Oral Liquid based on HPLC fingerprint and multi-components simultaneous determination / 中草药

Objective: To establish the HPLC fingerprint of liquid of raw material-intermediate product-Yinju Jiedu Oral Liquid (YJOL), and to determine the multi-components in YJOL, thus to provide an approach and basis for the quality control in production. Methods: The separation was performed on Shimadzu Inert Sustain C18 columin (250 mm × 4.6 mm, 5 μm) with mobile phase composed of acetonitrile-0.1% phosphoric acid solution (gradient elution) at the flow rate of 1.0 mL/min, the column temperature was set at 30 ℃ and the detection wavelengths were set at 240, 327, 334, and 280 nm. HPLC fingerprints of the extracts from 12 batches of raw material, intermediate product, and YJOL were established and compared. The content was determined and the common peaks were identified, and some of the characteristic peaks were analyzed. Results: Fifteen common peaks of intermediate product and liquid of raw material and 13 common peaks in YJOL were determined. The similarities of YJOL were over 0.9, the same with the similarities of the liquid of raw material and intermediate product. The correlation was good between the extracts from the liquid of raw material, intermediate product and YJOL. Peak 1 was from Isatidis Radix, peaks 2-7 were from Honeysuckle of Sichuan and Chrysanthemum indicum, peaks 8 and 9 were from Chrysanthemum indicum, peaks 10 and 11 were from Scrophulariae Radix, peaks 12 and 13 were from Scurfpea fruit. Based on the retention time of master compounds, six components [R, S-epigoitrin (peak 1), chlorogenic acid (peak 3), linarin (peak 10), harpagoside (peak 11), psoralen (peak 13), and psoralen (peak 14)] were identified and quantified. The average contents of six components were 39.21, 15.43, 2.14, 0.53, 7.21, and 6.51 mg/g in raw material, were 35.31, 11.54, 1.83, 0.37, 4.95, and 4.74 mg/g in intermediate product, and were 32.87, 10.58, 1.72, 0.31, 4.58, and 4.48 mg/g in YJOL. Conclusion: This validated method is suitable for the quality evaluation and quality control of YJOL.

Simultaneous determination of eight constituents in Compound Rhodiola Capsule by HPLC-UV-ELSD / 中草药

Objective: To develop an HPLC-UV-ELSD method for the simultaneous determination of crenelatin, gallic acid, salidroside, tyrosol, harpagide, harpagoside, angoroside C, and cinnamic acid in Compound Rhodiola Capsule. Methods: Kromasil C18 column (250 mm×4.6 mm, 5 μm) was adopted. The mobile phase was composed of acetonitrile (A) and 0.3% HAC (B) with gradient elution. The flow rate was 1.0 mL/min and the detection wavelength was 275 nm. the column temperature was 30℃, and the evaporative light-scattering detector (ELSD) drift tube temperature was 40℃, and gas pressure of 1.5 bar (150 kPa). Results: Crenelatin, gallic acid, salidroside, tyrosol, harpagide, harpagoside, angoroside C, and cinnamic acid were separated well. The linear calibration curves were obtained in 67.084-670.84 μg/mL for crenelatin, R2=0.9992; 11.410-114.100 μg/mL for gallic acid, R2=0.9994; 78.995-789.95 μg/mL for salidroside, R2=0.9996; 19.625-196.25 μg/mL for tyrosol, R2=0.9997; 59.368-593.68 μg/mL for harpagide, R2=0.9998; 62.585-625.85 μg/mL for harpagoside, R2=0.9995; 55.045-550.45 μg/mL for angoroside C, R2=0.9996; and 6.895-68.95 μg/mL for cinnamic acid, R2=0.9998. The average recoveries of the eight constituents were 100.8%, 98.9%, 100.1%, 100.8%, 98.9%, 99.6%, 100.7%, and 99.2% with RSD of 0.64%, 0.56%, 0.35%, 0.65%, 0.26%, 0.58%, 1.00%, and 0.64%. Conclusion: The method is convenient, accurate, and can be used for the simultaneous determination of the preparation.

Contents Determination of Harpagoside and Stilbene Glycoside in Shuangshen Xiaolong Granule by HPLC / 中国药房

OBJECTIVE:To establish a method for the contents determination of harpagoside and stilbene glycoside in Shuang-shen xiaolong granule. METHODS:HPLC of harpagoside was performed on the column of Kromasil 100-5 C18 with mobile phase of acetonitrile-1% acetic acid solution (gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 278 nm,column temperature was 25 ℃ and volume injection was 20 μl. HPLC of stilbene glycoside was performed on the column of Kromasil 100-5 C18 with mobile phase of acetonitrile-water(19∶81,V/V)at flow rate of 1.0 ml/min,etection wavelength was 320 nm,column temperature was 25 ℃ and volume injection was 10 μl. RESULTS:The linear range was 0.555 8-8.893 4 μg for harpagoside(r=0.999 9)and 0.010 6-0.340 2 mg for stilbene glycoside(r=0.999 6);RSDs of precision,stability and reproducibility tests were no more than 1.80%;recoveries were 97.30%-101.35%(RSD=1.43%,n=6) and 96.67%-100.83%(RSD=1.48%,n=6),respec-tively. CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the contents determination of harpa-goside and stilbene glycoside in Shuangshen xiaolong granule.

Simultaneous determination of harpagide and harpagoside in Mailuoning injection by HPLC / 中国生化药物杂志

Objective To establish a method for content determination of harpagide and harpagoside in Mailuoning injection by HPLC. Methods The experimental condition of HPLC method was as follows: SunfireTM C18 column (4.6 mm ×150 mm, 5 μm), with gradient elution using acetonitrile and 0.03% phosphoric acid; the detected wavelength was 210 nm, and the flow rate was 1.0mL/min.ResuIts Harpagide and harpagoside demonstrated good linear relationship in the range 0.1424~0.8544 μg/mL(r=0.9998) and 0.0732~0.4392μg/mL (r=0.9997) respectively.The average recovery rate were 98.22% and 99.27% with RSD of 1.46% and 1.42%(n=6)respectively.ConcIusion The method is simple, reliable, accurate, reproducible and stable, and it could be used in the determination of harpagide and harpagoside in Mailuoning injection.

Determination of eight active components in Xuanmai Ganjie Granules by UPLC-MS/MS / 中草药

Objective: To develop a UPLC-MS/MS method for simultaneously determining harpagide, liquiritin, harpagoside, platycodin D, ammonium glycyrrhetate, ophiopogonin D, methylophiopogonanone A, and methylophiopogonanone B in Xuanmai Ganjie Granules (composed with Scrophulariae Radix, Ophiopogonis Radix, Glycyrrhizae Radix et Rhizoma, and Platycodonis Radix) from different pharmaceutical companies. Methods: The chromatographic separation was achieved on Phenomenex Kenetix C18 column (50 mm × 2.1 mm, 5 μm) by using a mobile phase consisted of acetonitrile and 0.1% formic acid water at the flow rate of 0.3 mL/min for gradient elution. Simultaneous monitoring of positive and negative ions and multiple reaction monitoring (MRM) scan mode were applied to the quantification of the components in Xuanmai Ganjie Granules; Sample volume was 5 μL. Results: There was good linearity between the absorption peak area and the concentration for harpagide, liquiritin, harpagoside, platycodin D, ammonium glycyrrhetate, ophiopogonin D, methylophiopogonanone A, and methylophiopogonanone B in the ranges of 9-2250, 8-2000, 3.4-850, 96-24000, 12.4-3100, 3.6-1900, 1.7-425, and 1.5-375 ng/mL, respectively. The average recoveries were ranged from 97.2% to 102.8% (RSD ≤ 2.7%). The contents of harpagide, liquiritin, harpagoside, platycodin D, ammonium glycyrrhetate, ophiopogonin D, methylophiopogonanone A, and methylophiopogonanone B in eight batches of samples were in the ranges of 32.8-107.6, 54.8-178.0, 14.6-70.7, 31.2-280.0, 106.4-287.9, 0.1-0.6, 0.01-0.07, and 0.03-0.17 μg/g, respectively. Conclusion: The developed method is simple, effective, and credible for determining the eight components in Xuanmai Ganjie Granules. It provides more helpful information for the comprehensive quality evaluation of Xuanmai Ganjie Granules.

OPTIMIZATION OF EXTRACTION PROCESS OF SIMIAOYONGAN DECOCTION BY OR-THOGONAL TEST / 现代医院

Objective To research the best extracting technology of simiaoyongan decoction.Methods As the total contents of chlorogenic acid and harpagoside to be evaluation indicator, adopted L9 (34 ) orthogonal design to select out the best extraction.The main determination was by HPLC method.Results Accounted by the total contents of the two composi-tions, the influential effects of the experimental results were extraction solvent >reflux times >the volume of extraction>the circumfluence time.Conclusion The best extracting technology is adding 10-fold times of 50% alcohol, percolating 1 hours, and three times.

Study on Quality Standard for Compound Heishen Pills / 中国药师

Objective: To establish the quality standard for compound Heishen pills. Methods: Scrophulariae Radix, Radix et Rhizoma and Belamcancae Rhizoma were identified by TLC. HPLC was used to determine the content of harpagoside and cinnamic acid in Scrophulariae Radix on a Welchrom-C18 column using methanol-acetonitrile-1% ethylic acid (8∶21∶71) as the mobile phase. The flow rate was 1. 0 ml·min-1 . The column temperature was at 30℃ and the detection wavelength was set at 278 nm. Results:The TLC method had good specificity without interference from negative control. The linear range of harpagoside was 1. 32-65. 80μg·ml-1 with the average recovery of 98. 06%(RSD=2. 16%),and that of cinnamic acid was 0. 38-19. 20 μg·ml-1 with the average recovery of 98. 78%(RSD=1. 34%). RSDs of precision, stability and reproductibility tests were all below 2%. Conclusion: The established method is accurate, feasible and reproducible. It can be used in the quality control of compound Heishen pills.

Simultaneous Determination of Three Main Contents in Scrophulariae Radix Dispensing Particles by HPLC-UV Dual Wavelength Spectrometry Method / 中国中医药信息杂志

Objective To construct a simultaneous HPLC-UV dual wavelength spectrometry method for detecting three main contents - harpagide, harpagoside and cinnamic acid in Scrophulariae Radix dispensing particles. Methods Ultimate AQ-C18 column (250 mm×4.6 mm, 5μm) was used, with 1%acetic acid solution and acetonitrile as mobile phase of gradient elution. The flow rate was 1.0 mL/min, detection wavelength was 210 nm at former 13 min and 278 nm after 13 min. The column temperature was 30 ℃. Results The linear range of harpagide, harpagoside and cinnamic acid was 0.066 54-0.665 4 μg (r=1.000 0), 0.024 23-0.242 3 μg (r=0.999 9), and 0.100 28-1.002 8 μg (r=0.999 9), respectively. The average recovery (n=6) was 99.80%±1.22%, 100.31%±1.30% and 100.22%±1.24%, respectively. Conclusion The method is simple, repeatable, stable, and can be used for quality control and standardization of Scrophulariae Radix dispensing particles.