Multi-component content determination of Dracocephalum tanguticum by quantitative analysis of multi-components by single-marker / 中国中药杂志

This study aims to establish a method for the simultaneous determination of 7 active components in Dracocephalum tanguticum and to evaluate the quality of medicinal materials from different habitats. The method was established with high performance liquid chromatography(HPLC) and the gradient elution was performed with the mobile phase of acetonitrile-methanol-0.2% phosphoric acid solution at a column temperature of 35 ℃, an injection volume of 15 μL, and a flow rate of 0.6 mL·min~(-1). The detection wavelength was set as 215 nm. With rosmarinic acid as the internal reference, the relative correction factors and the content of other 6 components were calculated. The results were compared with those obtained with the external standard method. The results showed that the samples from Huangzhong county, Qinghai province had the best quality, with the highest content of p-hydroxybenzoic acid, cosmosiin, rosmarinic acid, oleanolic acid, and ursolic acid(9.29, 12.14, 6.02, 3.11, 17.67 mg·g~(-1) respectively). The samples from Chaya county, Tibet autonomous region ranked the second, with the highest content of betulin and betulinic acid(15.53, 7.17 mg·g~(-1), respectively). The method is accurate, reliable, and repeatable and suitable for the simultaneous determination of multiple components in D. tanguticum. The content of functional components varied in the samples from different producing areas and can be used as the indicator for the quality evaluation of medicinal materials.

Simultaneous determination of seven different components in Cynomorium songaricum Rupr. by QAMS / 药学学报

A quantitative analysis of multi-components by single marker method (QAMS) was established for simultaneous determination of gallic acid, protocatechuic acid, catechin, epicatechin, p-coumaric acid, ferulic acid and phloridzin in Cynomorium songaricum Rupr. The analysis was performed on a ChromCore Polae C18 column (250 mm×4.6 mm, 5 μm) , with a mobile phase consisting of acetonitrile-0.3% phosphoric acid aqueous solution for gradient elution. The volume flow rate, column temperature and sample injection volume were set at 1.0 mL·min-1, 25 ℃, and 40 µL, respectively. The relative correction factors of gallic acid and protocatechuic acid, catechin, epicatechin, p-coumaric acid, ferulic acid and phloridzin were calculated and the durability was also investigated. The contents of these seven compounds in fourteen batches of Cynomorium songaricum Rupr. from different producing areas or batches were determined by external standard method (ESM) and quantitative analysis of multi-components with a single-marker method (QAMS), respectively. SPSS and Origin Pro software were employed for principal components assay, similarity evaluation and cluster analysis. The specificity, precision, repeatability, stability and linear range (R2 > 0.999 0) of the seven components were all good. The average recovery was 96.89%-103.16% and RSD was 0.55%-2.76%. Then gallic acid was chosen as internal reference for calculation the correction factors for the other six components, the average relative correction factors of protocatechuic acid, catechin, epicatechin, p-coumaric acid, ferulic acid and phloridzin were 1.141 5, 0.200 5, 0.208 0, 2.361 9, 1.867 7, 0.204 6, respectively. Student's test results showed that there was no significant difference between the data analyzed by ESM and the data obtained from QAMS method. Through data visualization analysis, the contents of gallic acid, protocatechuic acid, catechin and epicatechin in different samples were significantly different, indicating that these four components might be the main quality markers of Cynomorium songaricum Rupr. for gaving more contributes to the principal components. The cluster analysis showed that samples from Xinjiang and samples from Inner Mongolia were clustered in significantly different categories, meaning that the quality of Cynomorium songaricum Rupr. had great relation with producing areas. The method of QAMS established in this study is a simple, economical and practical method with scientific and applicable charactistics for evaluating the quality of Cynomorium songaricum Rupr. efficiently and scientifically.

Quality evaluation of Aster souliei based on quantitative analysis of multi-components by single marker / 药学学报

To establish a quantitative analysis of multi-components by single marker (QAMS) for the determination of Aster souliei Franch., the relative correction factors (fx) of neochlorogenic acid, cryptochlorogenic acid, rutin, isoquercitrin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, quercetin, apigenin and kaempferol were established by ultra-high performance liquid chromatography with chlorogenic acid as internal reference. Meanwhile, the content of each component was determined by the external standard method (ESM) and QAMS, and a linear regression model was established to verify the feasibility and accuracy of the QAMS. Hierarchical clustering analysis (HCA) and orthogonal partial least square discriminate analysis (OPLS-DA) were used to evaluate the quality of 23 batches of A. souliei. The results showed that the repeatability of each fx was good. The average content of neochlorogenic acid, cryptochlorogenic acid, rutin, isoquercitrin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, quercetin, apigenin and kaempferol in 23 batches of A. souliei by QAMS was 0.165, 0.234, 6.115, 0.478, 0.484, 3.359, 1.382, 0.210, 0.172, and 0.057 mg·g-1, respectively. The mean content determined by the ESM method was 0.163, 0.235, 6.172, 0.479, 0.483, 3.343, 1.413, 0.207, 0.171, and 0.056 mg·g-1. The results of HCA and OPLS-DA analysis show that 23 batches of A. souliei can be divided into two groups based on caffeic acid content. The content of the first group was between 0.873 to 5.647 mg·g-1, while the second was between 8.524 to 16.705 mg·g-1. This QAMS method can be used to simply and quickly evaluate the quality A. souliei.

Evaluation on measurement uncertainty of correction factors of fluconazole impurities determined by HPLC standard curve method / 中国药科大学学报

@#In this paper, the uncertainties of correction factors of fluconazole impurities determined by HPLC standard curve method were evaluated, and the main common factors affecting the accuracy of standard curve method were found, so as to improve the accuracy of the method.In this study, the corresponding fitting lines of fluconazole and its impurities A, B, C, D, F and I were established respectively, and the ratio of the slope of fitting lines of each impurity and its corresponding principal component was calculated as the correction factor of the impurity.Then on the basis of GUM method, the uncertainty of each impurity correction factor determined by standard curve method was evaluated according to the established uncertainty evaluation scheme of correction factor determination process.The correction factor and uncertainty of fluconazole impurities A, B, C, D, F and I were 1.068 ± 0.046, 0.102 ± 0.005, 0.0582 ± 0.0031, 1.382 ± 0.121, 0.802 ± 0.067 and 1.383 ± 0.119, respectively, and the coverage factor k was 2.Finally, the contribution rate of each uncertainty component was calculated.In the relative combined standard uncertainties urel(f) of fluconazole impurities A, B, C, D, F and I correction factors, the sum of contribution rate of slope uncertainty urel(K) of the linear equation of principal component and its impurity is more than 85%; in the slope uncertainties urel(K) of linear equation, the contribution rates of uncertainties of solution concentration in 8 of 12 data groups are more than 80%, and the contribution rates of uncertainties introduced by reference substance content in solution concentration are about 80%.It can be seen that the preparation of linear solution concentration is the most influential factor in the determination of impurity correction factor by standard curve method, followed by the linear fitting process.In the preparation process of linear solution concentration, the purity of reference substance is the most influential factor, followed by weighing and pipetting times.The conclusion can help the experimenters to better formulate experimental plans and ensure the accuracy of the results when doing similar work.

Simultaneous determination of 10 components in Guanxinshutong capsules by quantitative analysis of multi-components by single marker / 药学学报

This study establishes a quantitative analysis of multi-components by single marker (QAMS) method for the simultaneous determination of gallic acid, sodium danshensu, protocatechuic acid, protocatechuic aldehyde, vanillin, rosmarinic acid, salvianolic acid B, eugenol, cryptotanshinone and tanshinone IIA in Guanxinshutong capsules (Bambusae Concretio Silicea, Salvia miltiorrhiza, clove, borneol, Bambusae Concretio Silicea) by HPLC. Sample was loaded onto an Agilent C18 (ZORBAX Extend-RP C18, 250 mm × 4.6 mm, 5 µm) column and eluted with methanol-0.4% aqueous formic acid solution as a flow phase gradient, flow speed 1.0 mL·min-1, detection wavelength 280 nm, column temperature 35 ℃ and sample intake of 5 µL. Using protocatechuic acid as the internal reference, a relative correction factor was calculated and the durability was investigated, and the content of 10 components was calculated by QAMS and external standard method (ESM). The results show that the specificity, linear relationship, precision, repeatability, and stability of the 10 components were good. The average recovery was 98.20%-103.47% and RSD was 1.26%-2.84%. The relative positive factors and contents of the other nine components were calculated as gallic acid (0.759, 227.381), sodium tanshinol (3.630, 3.283), protocatechualdehyde (0.185, 0.150), vanillin (0.532, 65.213), rosmarinic acid (4.240, 1.035), salvianolic acid B (3.245, 18.204), eugenol (1.729, 9.265), cryptotanshinone (0.691, 1.449), and tanshinone ⅡA (0.702, 1.939). The results of QAMS were consistent with ESM analysis, and the relative error was between -3% and 3%. This method is stable and reliable, and can be used for the determination of 10 components in Guanxinshutong capsules.

Content determination of five flavonoids in Tibetan medicine Rhododendron anthopogonoides by quantitative analysis of multi-components by single marker (QAMS) / 中国中药杂志

To establish a quantitative analysis of multi-components by single marker(QAMS) method for five flavonoids in Rhododendron anthopogonoides and verify its feasibility and applicability in the medicinal materials of R. anthopogonoides. With hyperoside as the internal reference, relative correction factors(RCF) of rutin, quercetin, quercitrin and kaempferol were established by high-performance liquid chromatography(HPLC) analysis. RCFs were used to calculate the content of each component, system durability and relative retention time. Simultaneously, QAMS and external standard method(ESM) were used to determine the content of five flavonoids in 12 batches of R. anthopogonoides from different origins. The results were statistically analyzed to verify the accuracy and feasibility. The fingerprints and cluster analysis data of R. anthopogonoides analyzed and discussed differences among the batches. According to the results, the RCFs of rutin, quercetin, quercetin and kaempferol in R. anthopogonoides were 1.242 6, 0.990 5, 0.535 0, and 0.781 3, respectively. The RCFs represented a good reproducibility under different experimental conditions. Besides, there was no significant difference between QAMS and ESM. Besides, the fingerprint and cluster analysis data showed the consistency between the classification and with the origin distribution of the herbs. In conclusion, the QAMS method shows a good stability and accuracy in the quality control of R. anthopogonoides.

Simultaneous Determination of 11 Active Components in Yuhuai Tablets by HPLC-QAMS / 中国药房

OBJECTIVE：To establish the method for simultaneous determination of 11 active components in Yuhuai tablets ， such as gardoside ，shanzhiside，gardenoside，genipin 1-gentiobioside，geniposide，ziyuglycoside Ⅰ ，ziyuglycoside Ⅱ ，narirutin， naringin，hesperidin and neohesperidin. METHODS ：HPLC-QAMS method was adopted. The determination was performed on Agilent TC-C 18column（250 mm×4.6 mm，5 μm）with mobile phase consisted of acetonitrile （A）-0.1％ phosphoric acid solution （B） （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃. The detection wavelengths were set at 238 nm for gardoside ，shanzhiside，gardenoside，genipin 1-gentiobioside and geniposide ，203 nm for ziyuglycoside Ⅰ and ziyuglycoside Ⅱ，and 283 nm for narirutin ，naringin，hesperidin and neohesperidin. Using geniposide as an internal reference ，the relative correction factors of other 10 components relative to this component were calculated ，and the contents of each component in 10 batches of samples were calculated. The results obtained by HPLC-QAMS method were compared with those obtained by external standard method. RESULTS ：The linear ranges of gardoside ，shanzhiside，gardenoside，genipin 1-gentiobioside，geniposide， ziyuglycoside Ⅰ，ziyuglycoside Ⅱ，narirutin，naringin，hesperidin and neohesperidin were 0.87-43.50，1.99-99.50，4.06-203.00， 7.35-367.50，12.97-648.50，28.98-1 449.00，3.79-189.50，1.57-78.50，18.05-902.50，0.66-33.00 and 14.38-719.00 μg/mL（all r＞0.999 0）. RSDs of precision ，repeatability and stability （24 h）tests were all less than 2％（n＝6）. The average recoveries were 96.90％-100.10％，and RSDs were 0.67％-1.74％（n＝9）. E-mail：289931673@qq.com There was no significant difference in the contents of 10 active components as gardoside between HPLC -QAMS method and external standard method in 10 batches of Yuhuai tablets （P＞0.05）. CONCLUSIONS ：The HPLC-QMAS method established in this study is convenient and accurate. It can be used for the simultaneous determination of gardoside ，shanzhiside，gardenoside，genipin 1-gentiobioside，geniposide，ziyuglycoside Ⅰ，ziyuglycoside Ⅱ，narirutin，naringin，hesperidin and neohesperidin in Yuhuai tablets.

Quality Control of Salvianolic Acids Reference Extract / 中国实验方剂学杂志

Objective::To establish the quality control method for multi-index content determination and fingerprint of salvianolic acids. Method::Agilent ZORBAX SB-C18 (4.6 mm×250 mm, 5μm) column was adopted, with 0.1%formic acid-water as mobile phase A and 0.1%formic acid-acetonitrile as mobile phase B for gradient elution (0-30 min, 20%-21.5%B; 30-35 min, 21.5%-25%B; 35-45 min, 25%-40%B; 45-50 min, 40%-95%B). The column temperature was set at 30 ℃, the flow rate was set at 1 mL·min-1, and the detection wavelength was set at 288 nm. Relative correction factors of caffeic acid, salvianolic acid E, rosmarinic acid, lithosperic acid, salvianolic acid B and salvianolic acid Y were determined by the concentration method. The content of each indicator component of the reference extract of salvianolic acid polyphenolic acid was determined and compared with the results of the monomer reference substance by the external standard method. At the same time, the fingerprint method was established. and the similarity evaluation was carried out on 10 batches of extracts. Result::Caffeic acid, salvianolic acid E, rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid Y had a good linear relationship within the respective detection mass concentration ranges (r>0.999 9). The injection precision RSD was 0.1%-1.2%, the reproducible RSD was 1.2%-1.6%, and the recovery of the six components was 82.03%-98.68%. The stability of each component in the sample solution was good within 36 h. The relative correction factors for each indicator component were determined to be caffeic acid (2.92), salvianolic acid E (1.10), rosmarinic acid (1.61), lithosperic acid (1.07), salvianolic acid B (1.00), salvianolic acid Y (0.83). The effects of different methods, concentrations, instruments, columns, wavelengths were investigated, and the measured relative correction factors were found to be suitable. The results of the calibration factor method and the monomer standard reference substance method were less different. The HPLC fingerprints of the reference extract of salvianolic acids were established, and five common characteristic peaks were determined. The chromatographic peaks were confirmed according to the reference substance. The similarity of the fingerprints of the 10 batches of extracts was higher, and the quality difference was smaller. Conclusion::The multi-index content determination method and the fingerprint method established in this study are simple, rapid, accurate and reproducible, and can be used for quality control of Salviae miltiorrhizae Radix et Rhizoma polyphenolic acid reference extract.

Exploration of Combinational Quality Control Method of Carbonized Ginger Based on HPLC Fingerprint and QAMS / 中国实验方剂学杂志

Objective::To establish the HPLC fingerprint of carbonized ginger and to determine the contents of zingerone, 6-gingerol, 6-shogaol, 10-gingerol, 8-shogaol and 10-shogaol with quantitative analysis of multi-components by single marker (QAMS). Method::The fingerprint of carbonized ginger was established by HPLC. All samples were analyzed by Waters SymmetryShield™ RP18 column (4.6 mm×250 mm, 5 μm) with gradient elution by acetonitrile(A)-water(B) (0-30 min, 25%-70%A; 30-50 min, 70%-90%A; 50-60 min, 90%A), the flow rate was 1.0 mL·min-1, the detection wavelength was set at 240 nm and the column temperature was 30 ℃. Zingerone, 6-gingerol, 8-gingerol, 6-shogaol, 10-gingerol, 8-shogaol and 10-shogaol was chosen as marker ingredients to establish HPLC fingerprint of carbonized ginger decoction pieces. Taking 6-gingerol as internal reference standard, the contents of zingerone, 6-shogaol, 10-gingerol, 8-shogaol and 10-shogaol were determined at the detection wavelength of 220 nm and 280 nm according to the relative correction factor. Result::The HPLC fingerprint of carbonized ginger was obtained and 10 common peaks were designated, and 7 of them were identified as zingerone, 6-gingerol, 8-gingerol, 6-shogaol, 10-gingerol, 8-shogaol and 10-shogaol, respectively. And there were no significant differences between the quantitative results of external standard method and QAMS. It is suggested that the content limits of carbonized ginger should be not less than 0.020%of zingerone (C11H14O3), 0.050%of 6-gingerol (C17H26O4), 0.120%of 6-shogaol (C17H24O3), 0.080%of 10-gingerol (C21H34O4), 0.030%of 8-shogaol (C19H28O3) and 0.050%of 10-shogaol (C21H32O3) calculated with reference to the dried products, respectively. Conclusion::The developed method is accurate and feasible, which can provide a simple and effective method for the quality control of carbonized ginger.

Determination of Diketo Aldehyde in Dihydroartemisinin Bulk Drug by Principal Component Reference Substances External Standard Method with Correction Factor / 中国药学杂志

OBJECTIVE: To establish a principal component reference substances external standard method with correction factor for the determination of impurity diketoal dehyde (DKA) in diketo aldehyde(DHA) bulk drug by selecting suitable liquid phase conditions. METHODS: The chromatographic C18 column (4.6 mm×250 mm, 5 μm) was used; isometric elution was set at conducted using acetonitrile-water (37∶63) as elution condition; the flow rate was 1 mL•min-1; the detection wavelength was 216 nm; the column temperature was maintained at 15 ℃; the injection volume was 20 μL. RESULTS: The DHA α peak and its impurity DKA were well separated. The average correction factor was 0.256 determined by three different chromatographic columns. CONCLUSION: The correction factor of DKA in DHA is accurate and reliable. This method can be used for the quality control of DKA in DHA raw materials.

Application of an HPLC correction factor method in pharmaceutical analysis / 药学学报

With the in-depth study of related substances and the development of consistency evaluation of generic drugs, relative correction factors are gaining increasing attention. By analyzing the domestic and foreign literature on correction factors in recent years, this paper describes the correction factor component, the current measurement method and its application. The rules and key points of use of an impurity correction factor and its determination and application are described, and some problems in its determination and application are discussed, providing a reference and basis for the standardization of research on impurity correction factors in the future.

Prevalence of familial hypercholesterolemia in premature coronary artery disease patients admitted to a tertiary care hospital in North India

Aims The prevalence of premature coronary artery disease (CAD) in India is two to three times more than other ethnic groups. Untreated heterozygous familial hypercholesterolemia (FH) is one of the important causes for premature CAD. As the age advances, these patients without treatment have 100 times increased risk of cardiovascular (CV) mortality resulting from myocardial infarction (MI). Recent evidence suggests that one in 250 individuals may be affected by FH (nearly 40 million people globally). It is indicated that the true global prevalence of FH is underestimated. The true prevalence of FH in India remains unknown. Methods A total of 635 patients with premature CAD were assessed for FH using the Dutch Lipid Clinical Network (DLCN) criteria. Based on scores, patients were diagnosed as definite, probable, possible, or no FH. Other CV risk factors known to cause CAD such as smoking, diabetes mellitus, and hypertension were also recorded. Results Of total 635 patients, 25 (4%) were diagnosed as definite, 70 (11%) as probable, 238 (37%) as possible, and 302 (48%) without FH, suggesting the prevalence of potential (definite + probable) FH of about 15% in the North Indian population. FH is more common in younger patients, and they have lesser incidence of common CV risk factors such as diabetes, hypertension, and smoking than the younger MI patients without FH (26.32% vs.42.59%; 17.89% vs.29.44%; 22.11% vs.40.74%). Conclusion FH prevalence is high among patients with premature CAD admitted to a cardiac unit. To detect patients with FH, routine screening with simple criteria such as family history of premature CAD combined with hypercholesterolemia, and a DLCN criteria score >5 may be effectively used.

Determination of seven flavonoids in Qingre Zhike Granules and intermediate product with quantitative analysis of multi-components by single marker / 中国中药杂志

This study is to determine seven flavonoids( narirutin,naringin,hesperidin,neohesperidin,baicalin,oroxylin A-7-glucoronide and wogonoside) in Qingre Zhike Granules and intermediate products by quantitative analysis of multi-components with a single marker( QAMS). High performance liquid chromatography( HPLC) was performed on Thermo BETASIL C18 chromatographic column( 4.6 mm×250 mm,5 μm) with acetonitrile-0.1% formic acid solution as mobile phase for gradient elution at a flow rate of 0. 8 m L·min-1. The detection wavelength was set at 280 nm and the column temperature was 30 ℃. Six relative correction factors( RCFs)of narirutin,naringin,hesperidin,baicalin,oroxylin A-7-glucoronide and wogonoside were established in the HPLC method with neohesperidin as the internal standard,and then these RCFs were used to calculate the mass fractions of the six components. Such mass fractions were compared with the mass fraction determined by the external standard method( ESM) at the same time to verify the feasibility and accuracy of QAMS method. Within the linear range,the RCFs of narirutin,naringin,hesperidin,baicalin,oroxylin A-7-glucoronide and wogonoside were 1.150,1.109,1.110,0.658,0.723 and 0.572,respectively,with good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method. The QAMS method with neohesperidin as the internal standard is feasible and accurate for the determination of narirutin,naringin,hesperidin,baicalin,oroxylin A-7-glucoronide and wogonoside,and it can be used for quality control of Qingre Zhike Granules and intermediate products.

Determination of 9 isoflavonoids in Puerariae Lobatae Radix with quantitative analysis of multi-components by single marker / 中国中药杂志

A method for determination of 9 isoflavones in Puerariae Lobatae Radix was established and the accuracy and feasibility of the method were verified. The relative correction factors of eight isoflavonoids,3'-hydroxy puerarin,puerarinapioside,3'-methoxy puerarin,puerarin 6″-O-xyloside,daidzin,genistin,formononetin and daidzein were determined by HPLC method with puerarin as the internal standard. The contents of 9 isoflavonoids in 11 batches of samples were determined by external standard method and QAMS.The accuracy and feasibility of the methods were evaluated by comparison of the quantitative results between external standard method and QAMS. The reproducibility of the relative correction factors was good under different experimental conditions,and there was no significant difference between the external standard method of the 9 compounds and the content of QAMS method. The results showed that using puerarin as an internal standard to simultaneously determine the 8 isoflavonoids mentioned above is accurate and feasible. Thus,it can be used as quality control of Puerariae Lobatae Radix.

Analysis on “Point-line-surface” Quality Standard of Yangjing Zhongyu Tang / 中国实验方剂学杂志

Objective: Based on the analysis of the total components of fingerprint and the determination methods of the existing components in the 2015 edition of Chinese Pharmacopoeia, the "point-line-surface" quality standard of Yangjing Zhongyu Tang was established by the "point" of each single component (morroniside, loganin, paeoniflorin, ferulic acid and verbascoside) to the "line" of multicomponent and the "face" of fingerprint of the whole component. Method: XB-C18 column (4.6 mm×250 mm, 5 μm) was used for gradient elution of 0.1% phosphoric acid aqueous solution-acetonitrile. The column temperature was 30℃, the injection volume was 10 μL, the flow rate was 1.0 mL·min-1, and the detection wavelengths were 240, 316, 230, 334 nm. The contents of these five components in Yangjing Zhongyu Tang were determined by three correction methods, external standard method and regression equation method. At the same time, the fingerprint of Yangjing Zhongyu Tang were analyzed by total component analysis and similarity evaluation. Result: With ferulic acid as reference, the relative correction factor (f) of morroniside, loganin, paeoniflorin and verbascoside were 0.392 1, 0.421 4, 0.261 7, 0.268 6 by multi-point correction method, and their f (slope correction method) were 0.385 4, 0.419 4, 0.255 9, 0.274 0, respectively. Twenty characteristic peaks of fingerprint were analyzed and the similarity was ≥ 0.999.There was no significant difference in the contents of these five components from Yangjing Zhongyu Tang determined by the quantitative assay of multi-components by single-marker (QAMS) correction method, the external standard method and the regression equation method. Conclusion: The comprehensive quality standard established by the total component analysis of fingerprint combined with various determination methods of existing components in the 2015 edition of Chinese Pharmacopoeia has been validated in famous classical formula of Yangjing Zhongyu Tang, which can provide ideas and methods for the quality control with quantitative determination and fingerprint of other famous classical formulas.

Simultaneous Determination of 4 Kinds of Effective Components in Volatile Oil of Forsythia suspensa by QAMS / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of α-pinene， β-pinene， limonene and α-terpineol in volatile oil of Forsythia suspensa. METHODS： GC method was adopted. The determination was performed on HP-5 capillary column through temperature-programmed route. The inlet temperature was 230 ℃， and detector temperature was 250 ℃； split sampling was applied （split ratio of 8 ∶ 1）； the air flow rate was 300 mL/min， the hydrogen flow rate was 30   mL/min， the tail gas flow rate was 30 mL/min， and the injection volume was 1 μL. Using limonene as internal reference， relative correction factors of α-pinene， β-pinene and α-terpineol were established， and the reproducibility of relative correction factors were investigated by using different chromatographs and columns， and chromatographic peak location of components was measured. The contents of above components were calculated with QAMS， and then compared with the results of external standard method. RESULTS： The linear range of α-pinene， β-pinene， limonene and α-terpineol were 16.5-990.0， 38.1-2 287.5， 8.2-491.2， 2.4-142.5  μg/mL， respectively （r≥0.999 1）. RSDs of precision， reproducibility and stability tests were all lower than 3％ （n＝6）. Average recoveries were 99.7％-105.5％（RSD＜4％，n＝9）. Compared with limonene （1.00），the average relative correction factors of α-pinene， β-pinene and α-terpineol were 0.91，0.86 and 1.11（n＝3）； relative retention time were 0.69-0.74， 0.81-0.86， 1.25-1.35（RSD＜3％，n＝3）. By using different chromatographs and columns， RSDs of relative correction factors were 0.21％-4.65％（n＝6）. Compared with external standard method， determination results of above 4 components were consistent （the absolute value of relative error were all less than 7％）. CONCLUSIONS： QAMS can be used for simultaneous determination of 4 kinds of effective components in volatile oil from F. suspensa.

Determination of four essential oils in cinnamomum cassia by quantitative analysis of multi-components by single marker / 中国药学杂志

OBJECTIVE: To establish a method of quantitative analysis of multi-components by single marker (QAMS) for determining four essential oils (cinnamaldehyde, cinnamyl alcohol, cinnamic acid, 2-methoxy cinnamaldehyde) in Cinnamomum cassia, and provide the experimental base for establishing the quality standard of Cinnamomum cassia. METHODS: Cinnamaldehyde was used as the internal reference standard, and the relative correction factors (RCF) of cinnamyl alcohol, cinnamic acid, and 2-methoxy cinnamaldehyde in Cinnamomum cassia were calculated. The contents of the four components were determined by both external standard method and QAMS. The validity of the QAMS method was evaluated by comparison of the quantitative results of both methods. RESULTS: The RCFs had good reproducibility, relative correction factor 0.673, 0.605 and 1.943, with RSDs of 0.529%, 0.373%, and 0.759%, respectively. No significant differences were found in the quantitative analysis results of cinnamyl alcohol, cinnamic acid, 2-methoxy cinnamaldehyde by using RCF and ESM. CONCLUSION: In the absence of reference substance, the content determination of the four essential oils in Cinnamomum cassia can be realized by QAMS, and this method can be used in the multi-index evaluation of Cinnamomum cassia essential oil constituents. It is suggested that the standard for cinnamaldehyde content be increased to 2.5%, and the contents of total cinnamyl alcohol, cinnamic acid and 2-methoxy cinnamaldehyde be not less than 0.2%.

Study on Substitute Reference Substance Method with DRS Origin Software: A Case Study of Determination of 11-keto-β-boswellic Acid and 11-keto-β-acetyl-boswellic Acid in Frankincense by UHPLC / 中国药学杂志

OBJECTIVE： To establish a UHPLC method for determination of the contents of 11-keto-β-boswellic acid(KAB) and 11-keto-β-acetyl-boswellic acid(AKBA)in Frankincense and explore the suitability and accuracy of substitute reference substance method with DRS origin software for qualitative and quantitative determination of chromatographic peaks. METHODS ：The samples were separated by UHPLC for determination of AKBA and KBA. AKBA was used as a reference to investigate the accuracy of KBA identification using DRS origin software on 19 different C18 columns. The RSDs of relative correction factors were calculated for different detection wavelengths and instruments.The relative correction factor method and the external standard method were selected for quantification and the differences were compared. RESULTS: The established method met the requirements of methodology and the average recovery was 100.21%(n=6) with RSD of 2.47%. The DRS origin software can be used to accurately determine the chromatographic peaks. The correct factor of AKBA vs. KBA was 0.936 and it was consistent under different conditions. There were no significant differences between the content calculated by the relative correction factor method and by the external standard method. CONCLUSION： This method is intelligent, feasible, reliable and economical, and can be used for the determination of frankincense content.

Establishment of National Reference Standards of Sulfadimidine Impurities / 中国药学杂志

OBJECTIVE: To establish the national reference standards of sulfadimidine impurities thus to provide guarantee for improving the standard of quality control of sulfadiazine in China. METHODS: First, the structures of sulfadimidine impurities A and E were validated by infrared spectrocopy, mass spectrum and nuclear magnetic resonance method. Then, the purities of impurities A and E were determined using the method of related substance test for sulphadiazine in the European Pharmacopoeia (version 9.0),their water content and residue on ignition were determined as well. The contents of sulfadimidine impurities A and E were determined by using mass balance method. Meanwhile, external standard method and nuclear magnetic quantitative method were used to calculate the content, which were mutually verified with the mass balance method. Finally, the correction factors of sulfadiazine impurities A and E at 241 nm were determined using standard curve method. RESULTS: The structures of sulfadimidine impurities A and E were confirmed, and the contents of impurities A and E were 99.1% and 98.7%, respectively, which were calculated by mass balance method. The results were consistent with those obtained from external standard method and nuclear magnetic quantification method. The correction factors of impurities A and E to sulfadimidine were 0.97 and 0.63, respectively. CONCLUSION: The first batch of national standard substances of sulfadimidine impurities A and E were established successfully.

Determination of Related Substance in Terazosin Hydrochloride Tablets by HPLC and Principal Component Self-control with Correction Factor / 中国药房

OBJECTIVE： To establish the method for the content determination of related substance in Terazosin hydrochloride tablets. METHODS： HPLC and principal component self-control with correction factor were adopted. The determination was performed on Agilent Zorbax Eclipse XDB C18 column with mobile phase consisted of acetonitrile-perchloric acid solution （20 ∶ 80， V/V） at the flow rate of 1.0 mL/min. The detection wavelength was set at 246 nm， and sample size was 20 μL. The column temperature was 50 ℃. The linear equations of terazosin hydrochloride， impurity A， B， C were drawn. The correction factors of each impurity related to terazosin hydrochloride were calculated by slope， and relative retention time was used to determine the position of impurities. The contents of impurity A， B and C in 3 batches of Terazosin hydrochloride tablets were determined and compared with the results of impurity control method. RESULTS： The relative retention time of impurity A， B， C was 0.39， 0.74， 2.77， respectively； the linear range of them were 0.25-3.0 μg/mL， respectively. The correction factors were 0.75， 1.09， 0.84， respectively. The detection limits were 0.35， 0.51， 0.43 ng， and the limits of quantification were 0.70， 1.02， 0.86 ng， respectively. The contents of impurity A， B and C in 3 batches of Terazosin hydrochloride tablets were 0.11％-0.13％， 0.03％ and 0.09％-0.12％； impurity B did not detected. The results are consistent with the determination of impurity control method. CONCLUSIONS： The method is simple， rapid and accurate for the content determination of related substances A， B， C in Terazosin hydrochloride tablets.