Quality evaluation methods of Poria based on yield and content of alkali extracted polysaccharide / 中国中药杂志

To improve the quality control methods of Poria and develop and utilize its resources fully, alkaline extraction was used in this study to determine the yield and content of alkali-soluble polysaccharides of Poria. The alkali-soluble extracts of Poria were obtained according to the optimum extraction conditions on the basis of single-factor test, and 30 batches of samples were determined. The structure and chemical composition of the alkali-soluble extracts was characterized by high-performance gel permeation chromatography(HPGPC), Fourier transform infrared spectrometry(FT-IR), nuclear magnetic resonance(NMR) spectroscopy and high-performance liquid chromatography(HPLC) with 1-phenyl-3-methyl-5-pyrazolone(PMP-HPLC). The results showed that the content of the alkali-soluble extracts was in the range of 46.98%-73.86%. The main component was β-(1→3)-glucan, and its molecular mass was about 1.093×10~5. Further, the content of alkali-soluble polysaccharides of Poria was measured by UV-Vis spectrophotometry and HPLC coupled with the evaporative light scattering detector(HPLC-ELSD), and 30 batches of samples were measured. The results indicated that the content of alkali-soluble polysaccharides determined by UV-Vis spectrophotometry was in the range of 73.70%-92.57%, and the content of samples from Hubei province was slightly higher than that from Yunnan province, Anhui province and Hunan province. The content of alkali-soluble polysaccharides determined by HPLC-ELSD was in the range of 51.42%-76.69%, and the samples from Hunan province had slightly higher content than that from the other three provinces. The content determined by UV-Vis spectrophotometry was higher than that by HPLC-ELSD. However, the content determined by HPLC-ELSD was close to that of alkali-soluble extract, which could accurately characterize the content of alkali-soluble polysaccharides in Poria, and the method was simple and repeatable. Therefore, it is recommended that the quantitative analysis method for alkali-soluble extract and alkali-soluble polysaccharides by HPLC-ELSD be used in the quality standards of Poria in Chinese Pharmacopeia.

Determination of five carbohydrate impurities in amino acid bulk drug by HPLC-ELSD / 中国药科大学学报

@#An analytical method was developed for the determination of five carbohydrate impurities in amino acid drug substances by high performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD). Sugar impurities in the amino acid sample were separated and enriched by cation exchange resin. A Lichropher NH2 column (4.6 mm × 250 mm, 5 μm) was used for chromatographic separation, and a gradient elution was performed using acetonitrile-water as mobile phase. The drift tube temperature was 40 oC, the gain value was 8, and nitrogen (350 kPa) was auxiliary gas. Method validation results showed that the limits of detection for fructose, glucose, sucrose, maltose and lactose were in the range of 20.8-75.0 mg/kg and that the limits of quantitation were in the range of 96.2-238.8 mg/kg. Good linear relationship (r ≥ 0.999) were in the linear range for the five sugars, and the recoveries ranged from 84.9%-107.8%. With easy operation, high sensitivity, good precision and reliable accuracy, the method can be used for analysis of residual sugar impurities in amino acid drug bulk drug.

Simultaneous Determination of 6 Carbohydrate Related Substances in Glucose by HPLC-ELSD / 中国药房

OBJECTIVE：To establish the m ethod for the simultaneous determination of 6 carbohydrate related substances in glucose as fructose ，maltose，isomaltose，maltotriose，maltotetraose and maltopentaose. METHODS ：HPLC-ELSD was adopted. The determine was performed on XBridge Amide column with mobile phase consisted of acetonitrile-water （75∶25，V/V）at a flow rate of 0.5 mL/min. The column temperature was set at 30 ℃，and the sample size was 10 L. The detector was evaporative light scattering detector ，the carrier gas was nitrogen ，the gas pressure was 40 psi，the evaporation temperature was 80 ℃，the drift tube temperature was 80 ℃，and the gain was 100. RESULTS ：The linear range of 6 carbohydrate related substances were 5.99-59.88， 9.90-98.96，9.92-99.19，5.97-59.74，4.03-40.32，5.89-58.89 μg/mL（r＞0.999 0）. The quantitation limits were 1.5，1.5，1.5，3.0， 3.0 and 3.0 μg/mL，respectively. The detection limits were 0.5，0.5，0.5，1.0，1.0，1.0 μg/mL，respectively. RSDs of precision ， stability（12 h）and reproducibility tests were all lower than 2.0％. The average recoveries were 95.87％-98.59％（RSD＝1.04％，n＝ 9），95.66％-99.84％（RSD＝1.20％，n＝9），96.11％-98.97％（RSD＝1.04％，n＝9），95.06％-99.11％（RSD＝1.25％，n＝9）， 95.69％-98.22％（RSD＝0.83％，n＝9），95.34％-98.56％（RSD＝1.01％，n＝9）. The contents of 6 carbohydrate related substances in 9 batches of glucose were 1.26-2.22，2.55-3.36，2.37-3.37，1.28-2.01，0-2.11 and 0-1.89 mg/g，respectively. CONCLUSIONS ： Established method is accurate and sensitive ，and can be used for the detection of carbohydrate related substances in glucose.

Analysis on Quality Evaluation Methods of Asparagi Radix Decoction Pieces and Its Standard Decoction / 中国实验方剂学杂志

Objective:To establish the quality evaluation methods of Asparagi Radix decoction pieces and its standard decoction. Method:Ten batches of Asparagi Radix standard decoction were prepared. High performance liquid chromatography-evaporative light scattering detection method (HPLC-ELSD) was established for the determination of protodioscin and protoneodioscin in Asparagi Radix decoction pieces and its standard decoction, and the fingerprint detection of Asparagi Radix decoction pieces with acetonitrile-water as mobile phase for gradient elution. UHPLC-LTQ-Orbitrap-MS/MS was used to identify ten main common peaks in the fingerprint with acetonitrile-0.1% formic acid solution as mobile phase for gradient elution, electrospray ionization (ESI) and positive and negative ion mode scanning were employed, the detection range was m/z 100-1 400. Result:The total content of protodioscin and protoneodioscin in Asparagi Radix decoction pieces was 0.41%-0.72%, and their total content in Asparagi Radix standard decoction was 0.33%-0.59%, the transfer rate of these two components was 73.6%-98.3%. The dry extract yield of the standard decoction was 59.0%-73.0%, and its pH was 4.9-5.6. There were 10 common peaks in the fingerprint, and all of them were saponins, including protoneodioscin, protodioscin, aspacochioside A and its isomer, methyl protodioscin, asparagoside F, (25R)-26-O-β-D-glucopyranosyl-furostan-5, 20-diene-3β, 26-diol-3-O-[α-L-rhamnopyranosyl (1→2)]-[β-D-glucopyranosyl (1→4)-α-L-rhamnopyranosyl (1→4)]-β-D-glucopyranoside, 26-O-β-D-glucopyranosyl-furostan-20 (22)-ene-3β, 26-diol-3-O-[α-L-rhamnopyranosyl (1→2)]-[α-L-rhamnopyranosyl (1→4)]-β-D-glucopyranoside, pseudodiosgenin, aspacochioside C. Conclusion:In this paper, the quality evaluation methods of Asparagi Radix decoction pieces and its standard decoction are established, and these methods are stable and feasible, which can provide reference for the quality control of pharmaceutical preparations containing Asparagi Radix.

Detectione of the contents of ginsenoside Rb1 and astragaloside IV in Weikang granules by HPLC-ELSD / 药学实践杂志

Objective To optimize the extraction method and develop the detection method of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules. Methods The extraction process of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules were optimized by single factor investigation, with the contents of ginsenoside Rb1 and astragaloside Ⅳ as optimization indicators. The HPLC-ELSD method was developed for the detection of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules. Separation was carried out on an XBridge®Shield RP18 column (4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-water（32:68）at the flow rate of 1 ml/min. The column temperature was maintained at 30 ℃. The drift tube temperature was set at 60 ℃, and the carrier gas flow rate was 1.7 SLM. Results The optimized extraction methods of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules were as the following: methanol reflux extraction for 1.5 h, and n-butanol extraction and ammonia washed for 5 and 2 times, respectively. The HPLC-ELSD method was established to detect the contents of ginsenoside Rb1 and astragaloside Ⅳ. The linear relationship was good (r > 0.9997). The intra-day and inter-day precision was less than 1%. The recovery rates were 95.65% and 100.57%. The stability and repeatability RSD were less than 3%. The contents were 2.8630 mg/g and 0.2576 mg/g. The RSDs were 0.62% and 1.51%, respectively. Conclusion The extraction method of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules is optimized, and a reliable, accurate and reproducible HPLC-ELSD method for the detection of the contents of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules is established.

Ingredients changes in Rehmanniae Radix processing based by HPLC-ELSD specific chromatogram / 中国中药杂志

To establish the HPLC-ELSD specific chromatogram analysis method of Rehmanniae Radix and Rehmanniae Radix Prae-parata, and analyze and compare their chemical compositions, so as to reveal the change regularity of compositions during the proces-sing. By HPLC-ELSD method, the chromatographic column for Prevail Carbohydrate ES(4.6 mm ×250 mm, 5 μm) was adopted, with acetonitrile(A)-water(B) as mobile phase for gradient elution, and the evaporative light-scattering detector was used. A total of 23 batches of Rehmannia Radix samples, and 25 batches of Rehmanniae Radix Praeparata samples and processing dynamic samples were compared. The established method had a great repeatability, precision and stability. Eight common chromatographic peaks were extracted from 23 batches of Rehmanniae Radix samples, 8 common peaks were extracted from 25 Rehmanniae Radix Praeparata, and 7 chromatographic peaks were identified. The composition ratio of Rehmannia Radix was changed greatly during the processing. When the simila-rity≥0.95 and the fructose peak area was more than 2 times of stachyose tetrahydrate or more than 20 times of raffinose, the processing degree conformed to the requirements of empirical identification. The three main oligosaccharides of Rehmanniae Radix were sucrose that was heated to generate fructose and glucose, stachyose tetrahydrate that was heated to generate melibiose, sucrose and fructose, and stachyose tetrahydrate that was heated to generate manninotriose. The change in the index of proportion between monosaccharides and oligosaccharides can be used as the quantitative criterion for the processing quality of Rehmanniae Radix Praeparata.

Qualitative and quantitative analysis of major cholic acids in Suis Fellis Pulvis / 中国中药杂志

This study is to establish a qualitative method for rapid identification of bile acids in Suis Fellis Pulvis based on UHPLC-LTQ-Orbitrap-MS technology,and an HPLC-ELSD internal standard method for the quantitative determination of two glycine-conjugated BAs in Suis Fellis Pulvis.The chromatographic separation of the UHPLC-LTQ-Orbitrap-MS qualitative analysis was achieved on a Waters Acquity UPLC HSS T_3column(2.1 mm×100 mm,1.8μm),with 0.2%formic acid aqueous solution(A)-acetonitrile(B)as mobile phase ingradient elution.Electrospray ionization(ESI)source was applied and operated in negative ion mode.Quantitative analysis was performed at 30℃on a Diamonsil-C_(18)column(4.6 mm×250 mm,5μm).The mobile phase consisted of 0.2%formic acid solution and acetonitrile with gradient elution and the flow rate was 1.0 m L·min~(-1).An ELSD was used with a nitrogen flow-rate of1.4 L·min~(-1)at a drift tube temperature of 60℃and the gain was 1.A total of 14 bile acids in Suis Fellis Pulvis were characterized based on the accurate mass measurements,fragmentation patterns,chromatographic retention times,and reference materials.For the quantitative analysis method,the glycohyodeoxycholic acid and glycochenodeoxycholic acid had good linear relationship in the range of26.52-265.20 mg·L~(-1)(r=0.999 8)and 19.84-198.40 mg·L~(-1)(r=0.999 1),respectively.The average recoveries(n=6)were104.1%and 103.1%,and the RSD were 2.0%and 2.4%.The UHPLC-LTQ-Orbitrap-MS technology provides a fast and efficient qualitative analysis method for identification of bile acids in Suis Fellis Pulvis.The HPLC-ELSD internal standard method is accurate and reliable,which has reference value for the quality control of Suis Fellis Pulvis.

Method Improvement for Content Determination of Astragaloside Ⅳ in Xiangju Granules and Its Consistency with the Com- ponents of Original Formulation / 中国药房

OBJECTIVE：  To improve the method for the content determination of astragaloside Ⅳ in Xiangju granules， and to evaluate the consistency of relevant preparations with the components of original formulation， so as to provide evidence for the modern preparation of TCM compound. METHODS： HPLC-ELSD method was established for the content determination of astragaloside Ⅳ in Xiangju granules， and compared with original standard TLC scanning. Using critrinin， ferulic acid， calycosin glucoside， liquiritin， glycyrrhizic acid， rosmarinic acid， buddleoside and magnoline as control， HPLC method was used to determine the release components of self-made Xiangju granules， Xiangju capsules， Xiangju tablets in water. Fingerprint characteristics chromatogram of different Xiangju preparations and original formulation extract were compared by using Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 version）. At the same time， HPLC-ELSD method was used to determine and compare the release rate of astragaloside Ⅳ from different Xiangju preparations and original formulation extract in water. RESULTS： Established HPLC-ELSD method was specific. The linear range of astragaloside Ⅳ was 0.13-2.10 mg/mL. RSDs of precision， repeatability and stability tests were all lower than 3％ （n＝6）， and average recovery was 97.66％ （RSD＝1.01％，n＝6）. Average content of astragaloside Ⅳ by this method was 0.398 mg/g （RSD＝1.01％， n＝3）， which had better reproducibility than TLC scanning. The comparative results of characteristic fingerprints showed that the similarity among Xiangju granules， Xiangju capsules， Xiangju tablets and the original formulation dry extract powder was more than 0.850. Average release rates of astragaloside Ⅳ in Xiangju granules， Xiangju capsules， Xiangju tablets and the original formulation extract were 0.392， 0.358， 0.349， 0.389 mg， respectively. Compared with original formulation extract， there was no statistical significance in release rate of astragaloside Ⅳ in Xiangju granules （P＞0.05）， while there was statistical significance in Xiangju capsules and Xiangju tablets （P＜0.01）. CONCLU- SIONS： Established HPLC-ELSD method is accurate and feasible， and is suitable for the content determination of astragaloside Ⅳ in Xiangju granules. The main components of Xiangju granules are consistent with original formulation.

Study on Quality Standard of Compound Platycodon grandiflorum Antitussive Tablets / 中国药房

OBJECTIVE： To establish the quality standard of Compound Platycodon grandiflorum antitussive tablets. METHODS： TLC was used to identify the P. grandiflorum， Polygala tenuifolia and Glycyrrhiza uralensis qualitatively in Compound P. grandiflorum antitussive tablets. HPLC-ELSD method was used to measure the content of platycodin D in Compound P. grandiflorum antitussive tablets. The determination was performed on Agilent C18 column with mobile phase consisted of acetonitrile-water （26 ∶ 74， V/V） at the flow rate of 1.0 mL/min. ELSD was used with drift tube temperature of 105 ℃， gas flow rate of 3.0 L/min and column temperature at 35 ℃. RESULTS： TLC chromatograms of P. grandiflorum， P. tenuifolia and G. uralensis had clear spots with good separation and no same spot from negative samples. The linear range of platycodin D was 0.421 9- 5.062 8 μg （r＝0.999 9）. The quantitative limit and detection limit were 0.364， 0.109 μg， respectively. RSDs of precision， stability， reproducibility and durability tests were all lower than 3.0％. The recovery rates were 87.32％-91.96％ （RSD＝1.73％，n＝6）. The platycodin D contents of 178 samples ranged from 0.004 to 0.73 mg/tablet. The content of platycodin D in 55 batches （30.9％） of samples was lower than the content limit （0.10 mg/tablet） proposed in this study. CONCLUSIONS： Established method is accurate and reliable， and can be used for the quality control of Compound P. grandiflorum antitussive tablets.

Contents Determination of Spinosin and Jujub oside A in the Seads of Ziziphus jujuba and Its Quality Grading Standard / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of spinosin and jujuboside A in the seads of Ziziphus jujuba， and to investigate its quality grading standard. METHODS： HPLC-ELSD method was adopted. The separation was carried out on Inertsil ODS-SP column with mobile phase consisted of acetonitrile-water （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃， the temperature of drift tube was 90 ℃， the flow of carrier gas was 2.9 L/min and injection volume was 20 μL. The thickness， width， length and 100-grain quality of the medicinal materials were used as indicators to investigate the appearance traits. SPSS 22.0 software was used to analyze the correlation of the contents of spinosin and jujuboside A， its appearance traits with the quality constant of TCM， and establish a quality classification standard for the seads of Z. jujuba. RESULTS： The linear range of spinosin and jujuboside A were 1.03-6.18 μg/mL （r＝0.999 7）， 1.05-6.30 μg/mL （r＝0.999 8）； the limits of quantitation were 0.171， 0.174 μg/mL， respectively； the limits of detection were 0.052， 0.053 μg/mL， respectively. RSDs of precision， stability and reproducibility tests were all lower 2％. The recoveries were 99.01％-102.97％ （RSD＝1.39％， n＝6）， 97.94％-101.03％ （RSD＝1.13％， n＝6）， respectively. Correlation analysis results showed that the length， width， 100-grain quality spinosin content and jujuboside A content of the medicinal materials were positively correlated with the quality constant of TCM. The results of quality classification for 30 batches of medicinal materials showed that S1-S4 and S7-S12 were first-class products； S5， S6， S13-S17 and S20-S30 were second-class products； S18 and S19 were third-class products. CONCLUSIONS： Established content determination method is simple， precision， accurate and stable， and can be used for simultaneous determination of spinosin and jujuboside A in the seads of Z. jujuba. Established quality grading standard of the seads of Z. jujuba can be used to evaluate the quality.

Simultaneous Determination of Seven Isosteroidal Alkaloids in Fritillaria wabuensis by HPLC-ELSD Method / 中国药学杂志

OBJECTIVE: To establish an effective high performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) method to simultaneously detect multi alkaloids in Fritillaria wabuensis, which can be used to control the quality of Fritillaria wabuensis. METHODS: The method was conducted on an InertSustain C18 column (4.6 mm×250 mm, 5 μm) with 0.1% TFA (V/V) aqueous solution-acetonitrile as mobile phase eluted in a gradient program. RESULTS: The method had good linearity (r2≥0.999 0).The accuracy was between 90% and 105%. This method was simple, sensitive, accurate and specific, which can be applied to determine imperialine-3-β-D-glucoside, imperialine, peimisine, verticine, verticinone, isoverticine and chuanbeinone in Fritillaria wabuensis. CONCLUSION: Nearly all fritillariae alkaloids can be detected under this condition, therefore, the chromatogram can be used as the characteristic chromatogram to testify different kinds of Fritillaria.The method can also be combined with mass spectrometric detector. It′s the first time for isoverticine and chuanbeinone to be detected in Fritillaria wabuensis by HPLC and also the first time for seven isosteroidal alkaloids in Fritillaria wabuensis to be simultaneously determined.

Simultaneous determination of eight components in Buxue Shengru Granules by HPLC-ELSD / 中草药

Objective: To establish an HPLC-ELSD method for simultaneous determination of astragaloside IV, ferulic acid, paeoniflorin, glycyrrhizic acid, vaccarin, naringin, neohesperidin, and platycodin D in Buxue Shengru Granules (BSG). Methods: The analysis of methanol extract of this drug was performed on a Diamonsil C18 (250 mm × 4.6 mm, 5 μm) with the mobile phase comprising of 0.1% formic acid aqueous solution-acetonitrile. Flow rate was 1 mL/min and column temperature was 30 ℃. The samples were tested on Evaporative Light-scattering Detector with drift tube temperature of 40 oC. Results: The analysis permitted very good separation of eight constituents within 52 min. The linearity ranges of the eight constituents were 62.5-1 250.0 μg/mL (r > 0.995 0). The average recoveries of eight constituents in the samples were in the range of 95.51%-99.47%, and the RSD ranged from 0.31% to 3.70%. Intraday and interday precisions RSD of the peak areas of the eight components were less than 3%; The repeatability RSD of each component ranged from 0.94% to 2.11%; Eight components had good stability within 18 h, and the concentration RSD of each component ranged from 0.98% to 2.86%. The content of vaccarin, paeoniflorin, ferulic acid, naringin, neohesperidin, platycodin D, astragaloside IV, and glycyrrhizic acid were 0.550 7-0.584 3, 19.657 9-19.952 1, 0.350 5-0.384 7, 18.794 7-19.557 3, 12.124 7-12.414 2, 0.610 7-0.631 3, 0.238 8-0.274 3, and 2.750 4-2.852 2 mg/g, respectively. Conclusion: This simple and accurate method can be used for the rapid quality control of BSG.

Determination of three saponins in Compound Shiwei Tablets and study on transfer rate in its preparation process / 中草药

Objective: High performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method was used to establish the determination method for the three kinds of saponins (astragaloside I, II, and IV) in Compound Shiwei Tablets (CST), and investigate the three saponins components’ s transfer rate in the preparation process of CST in order to improve the quality control method of CST. Methods: A HPLC-ELSD method was operated on the column of Agilent 5-HC C18 (2) (250 mm × 4.6 mm, 5 μm), with acetonitrile-water as the mobile phase for gradient elution, at a flow rate of 1.0 mL/min, with column temperature of 30 ℃ and injection volume of 20 μL. The ELSD parameters were as follow: the carrier gas flow rate was 1.5 L/min, the drift tube temperature was 90 ℃. Determinate the content of astragaloside I, II, and IV in products, granules and extracts of CST, and calculate the transfer rate of three saponins in the preparation process of CST. Results: A method for the determination of astragaloside I, II, and IV in CST was established. Under this condition, all three components reached baseline separation with good linear relationship. The average recovery rates were 99.58%, 99.31% and 99.51%, and RSD values were 3.0%, 2.5% and 2.5%, respectively. Astragaloside I had lower transfer rate during the preparation process, and the transfer rate of astragaloside IV was the higher in the preparation process, both of which were greater than 100%. Conclusion: This study established a method for simultaneous determination of three kinds of saponins of astragaloside I, II, and IV in CST. The method has good reproducibility and strong specificity, which is simple and easy,and can be used to inspect the transfer rates of three kinds of saponins in the preparation process and improve the quality control standard of saponins in CST, and provide reference for the quality control of other traditional Chinese medicine preparations containing astragalus.

Quality Control of Zhenqi Fuzheng Granule Based on Multi-component Determination / 中国实验方剂学杂志

Objective: To estimate the overall quality characteristics of Zhenqi Fuzheng granules (ZQFZ),which were composed of Ligustri Lucidi Fructus and Astragali Radix and collected from different manufacturers (their final preparations included two types,contained sugar and sugar free) by established HPLC methods,in order to propose an appropriate quality-control strategy for promoting the quality control specification of ZQFZ.Method: The quantification of the 6 components (rhodioloside,calycosin-7-O-β-D-glucoside,specnuezhenide,ononin,calycosin and astragaloside IV) were performed on a C18 column with two chromatographic systems.Chromatographic system Ⅰ:methanol and water were adopted as mobile phase with gradient elution,the flow rate was 1.0 mL·min-1,and optimum detection waves were at 224,250 and 275 nm respectively.Chromatographic system Ⅱ:methanol and water (80:20) were adopted as mobile phase with gradient elution at the flow rate of 1.0 mL·min-1,and the detector parameters were set as follows:the drift tube temperature was 75℃,and the carrier gas flow rate was 1.5 L·min-1.Both column temperatures were at 30℃.All of the 80 batches of ZQFZ from different manufacturers were determined and analyzed.Result: All of the six markers could be detected in 80 batches of ZQFZ,but their contents were quite different.The results of the one-way ANOVA showed significant differences between manufacturer 4 and other three manufacturers in sugar-containing preparations (P PConclusion: It is of great significance to increase relevant quality control markers of Ligustri Lucidi Fructus in ZQFZ,such as rhodioloside and specnuezhenide,for standardizing production and improving quality level.

Effect of fermentation on components of bile acids in Arisaema Cum Bile and determination of three kinds of free bile acids in Arisaema Cum Bile / 中国中药杂志

The aim of this study is to analyze the compositions of main bile acids in fermented and mixed processing products of arisame cum bile from pig bile, and to establish a method for content determination of bile acids in fermented Arisaema Cum Bile. Fermented and mixed processing products were prepared from arisaematis rhizome and arisaematis rhizoma preparatum with pig bile respectively. Then the differences in bile acids compositions between such two kinds of products were compared by high performance liquid chromatography and evaporative light-scattering detector (HPLC-ELSD). With three kinds of free bile acid compositions as the indicators, HPLC-ELSD method was adopted to determine the content of bile acid compositions in fermented product,on Agilent Eclipse XDB C₁₈(4.6 mm×250 mm, 5 μm) chromatographic column, with acetonitrile and 0.1% glacial acetic acid solution (55:45) as mobile phase, at a flow rate of 1 mL·min⁻¹, column temperature of 30 °C, drift tube temperature of 90 °C, and a nitrogen flow rate of 2.2 mL·min⁻¹. The results showed that the bile acids in fermented bile Arisaema were mainly in a free form, while in mixed processing product, the compositions were mainly in a conjugated form. Three kinds of free bile acids, namely porcine cholic acid (HCA), porcine deoxycholic acid (HDCA) and chenodeoxycholic acid (CDCA) in fermented product, showed a good linear relationship in the range of quantification. The average recovery rate was 95.99%-104.3%, complying with the requirements. The results showed that the conjugated bile acids could be transformed into free bile acids during the fermentation of arisaema cum bile. This established method can effectively control the content of bile acids compositions in fermenting arisaema cum bile.

Determination of three pair of triterpenoid isomers in leaf of Ilex hainanensis by HPLC-ELSD / 中国中药杂志

The present study is to develop an HPLC-ELSD method for simultaneous determination of three pairs of triterpenoid isomers, Ilexsaponin A₁, Ilexhainanoside D, Ilexgenin A, 3β, 19α-dihydroxyolean-12-ene-24, 28-dioic acid (ilexhainanin E) ursolic acid and oleanic acid in the leaf of Ilex hainanensis, which could provide evidence to the quality control of this herb. The six constituents were measured on a Waters XBridge C₁₈ column (4.6 mm×250 mm, 5 μm), with a mobile phase consisting of methanol (A)- 0.5% formic acid in water (B) at a flow rate of 1.0 mL·min⁻¹ (0-18 min，70%-85% A，18-20 min，85%-95% A；20-35 min，95% A). The carrier gas was N₂, and the pressure was 2.8 L·min⁻¹. The drift tube in this experiment were set at 70 °C. The injection volume was 10 μL. The contents of the six triterpenoids in 6 samples were 3.7-8.5, 10.3 -22.1, 2.8-5.9, 7.8-14.1, 2.6-3.8 and 8.8-11.9 mg·g⁻¹, respectively. The established method is proved to be accurate and sensitive for the determination of triterpenoids in Ilicis Hainanensis Folium, and may be used for the quality improvement of this herb.

Simultaneous determination of eight constituents in Shenmai Zhisou Syrup by HPLC-ELSD / 中成药

AIM To establish an HPLC-ELSD method for the simultaneous content determination of euscaphic acid,tormentic acid,crategolic acid,corosolic acid,ophiopogonin D,ophiopogonin D',methylophiopogonanone A and methylophiopogonanone B in Shenmai Zhisou Syrup (Eriobotryae Folium,Ophiopogonis Radix,Glehniae Radix,etc.).METHODS The analysis of methanol extract of this drug was performed on a 35 ℃ Alltima C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of (acetonitrile-methanol)-0.5％ ammonium acetate flowing at 0.7 mL/min in a gradient elution manner.RESULTS Eight constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 96.85％-99.43％ with the RSDs of 0.57％-1.33％.CONCLUSION This simple and accurate method can be used for the rapid quality control of Shenmai Zhisou Syrup.

Preparative separation and quantitative determination of two kaurenoic acid isomers in root barks of Acanthopanax gracilistylus / 中国天然药物

The kaurenoic acid-type diterpenoids in Acanthopanacis Cortex have been reported to be the major active components. However, the diterpenoids are present as position isomers that exacerbate the challenges in obtaining standards compounds. Little work has been done on the quantitative analysis of the diterpenoids in the herb. In the present study, two diterpenoid isomers ent-16βH,17-isovalerate-kauran-19-oic acid (1) and ent-16βH,17-methyl butanoate-kauran-19-oic acid (2) with high purity were separated by analytical HPLC, followed by recrystallization in acetone. Furthermore, an HPLC-ELSD method was developed and validated for simultaneous determination of 1 and 2 in 9 batches of Acanthopanacis Cortex samples. The HPLC separation and quantification was achieved in 40 min using an Agela Promosil C column eluted with a gradient of water and acetonitrile. The calibration curves showed good linearity (r ≥ 0.999 9) within the test ranges. The LOD ranged from 0.407 2 to 0.518 0 μg and LOQ ranged from 1.018 0 to 1.295 0 μg. The precisions (%RSD) were within 1.47% for the two isomers. The recovery of the assay was in the range of 98.78%-99.11% with RSD values less than 2.76%. It is the first time to establish a quantitative HPLC method for the analysis of the bioactive kaurenoic acid isomers in the herb.

Dissolution of Yinxingye Dispersible Tablets by Quantitative Analysis of Multi-components with Single-marker / 中国药师

Objective:To establish a quantitative analysis method for determining the dissolution of Yinxingye dispersible tablets by quantitative analysis of multi-components with single-marker ( QAMS) . Methods:A small glass method was adopted. Hydrochloric acid (0. 1 mol·L-1 ) was used as the dissolution medium, and the stirring speed was at 50 r·min-1 . HPLC-ELSD with gradient elu-tion combined with QAMS was used to simultaneously determine the dissolution of 3 total flavonoids ( artemisinin, quercetin and isorh-amnetin) and 4 total lactones (ginkgo esters and lactones a, b and c) in Yinxingye dispersible tablets. Results:Within a certain line-ar range, the relative correction factors (RCFs) of quercetin and isorhamnetin with the reference of artemisinin was 1. 873 and 0. 324, respectively, and the RCFs of lactones a, b and c with the reference of ginkgo esters was 2. 280, 1. 659 and 1. 429, respectively, and the repeatability was good under the different experimental conditions. There were no significant differences between the calculated val-ues by QAMS and those by the external standard method in 5 batches of Yinxingye dispersible tablets, and the results showed that the RCFs was authentic. The dissolution uniformity of Yinxingye dispersible tablets was good in 0. 1 mol·L-1 hydrochloric acid. Conclu-sion:The method is simple, accurate and reproducible in the dissolution determination of Yinxingye dispersible tablets.

HPLC-ELSD fingerprints of oligosaccharide sites from mycelia of Hericium erinaceum solid cultures and Weilening Tablets / 中成药

AIM To establish the HPLC-ELSD fingerprints of oligosaccharide sites from mycelia of Hericium erinaceum solid cultures and Weilening Tablets.METHODS The analysis of aqueous extract from samples was performed on a 80 ℃ thermostatic Waters XBridge TM Amide column (4.6 mm × 150 mm,3.5 μm),with the mobile phase comprising of acetonitrile-0.2％ ammonium acetate flowing at 1 mL/min in a gradient elution manner.RESULTS There were eight and nine common peaks in two HPLC-ELSD fingerprints with the similarties of 0.994-0.966 and 0.990-0.997,respectively.Three of them were mannitol,lactose and trehalose,which showed good linear relationships within their own ranges (r ≥ 0.999 0),the average recoveries were 95.08％-104.82％ with the RSDs of 1.12％-2.90％.CONCLUSION This simple and accurate method can be used for the rapid quality control of mycelia of Hericium erinaceum solid cultures and Weilening Tablets.