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  • Li-qiu SUN, Dan WANG, Ying-nan ZHAO, Zhi-chun SHI, Jun LI, Jin-lan WANG, Ming ZHAO, Shu-jun ZHANG
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 806-815.
    Objective:

    To establish a quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of 7 flavanoids (5-hydroxy-6,7,3’,4’-tetramethoxyflavone,apigenin,hispidulin,kaempferol,jaceosidin,eupatilin and casticin) in Artemisiae Argyi Folium.

    Methods:

    The HPLC system consisted of the Agilent ZORBAX SB-C18 column (150 mm×4.6 mm,5 μm) column with gradient elution of acetonitrile and 0.2% phosphoric acid as the mobile phase at a flow rate of 1.0 mL·min-1,a detection wavelength of 350 nm,and a column temperature of 30 ℃. Eupatilin was selected as the internal reference substance,the relative correction factors between eupatilin and the other 6 flavanoids were established,and the contents of these 7 constituents in samples were calculated to realize QAMS. At the same time,compared with the external standard method to verify the accuracy and feasibility of the QAMS method.

    Results:

    Within a certain linear range,the relative correction factors between eupatilin and 5-hydroxy-6,7,3’,4’-tetramethoxyflavone,apigenin,hispidulin,kaempferol,jaceosidin as well as casticin were 0.958,1.387,1.000,0.950,0.957 and 1.297,respectively (RSDs of RCFs were less than 2.0%). The contents of 5-hydroxy-6,7,3’,4’-tetramethoxyflavone,apigenin,hispidulin,kaempferol,jaceosidin,eupatilin,casticin in 20 batches of Artemisiae Argyi Folium were 0.031 4-0.623 5 mg·g-1,0.000 9-0.092 6 mg·g-1,0.020 6-0.170 7 mg·g-1,0.011 0-0.184 7 mg·g-1,0.011 7-0.864 0 mg·g-1,0.253 2-2.555 0 mg·g-1 and 0.015 6-0.250 7 mg·g-1,respectively.

    Conclusion:

    Using eupatilin as the internal reference,QAMS method for 7 flavanoids is established. The method is accurate and reliable,and can be used for quality control and quantitative analysis of Artemisiae Argyi Folium.

  • Sheng-yun DAI, Jie LIU, Su-ning YUN, Chao-jie LIAN, Fei QIAO, Ke ZAN, Li-nong GUO, Shuang-cheng MA, Jian ZHENG
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 740-749.

    The national drug sampling and inspection project is an important way of drug quality supervision in China which providing strong support for drug supervision and standard improvement. This article summarizes the national drug sampling and inspection project of Arnebiae Radix completed by Institute for Control of Chinese Traditional Medicine and Ethnic Medicine,National Institutes for Food and Drug Control in 2015 and 2022. The results illustrated that the qualification rate of Arnebiae Radix has increased from 43.9% in 2015 to 87.5%,and the qualification rate of Arnebiae Radix has significantly increased. The two nationwide inspections of Arnebiae Radix reflected the scarcity of Arnebiae Radix resources in Xinjiang and Inner Mongolia,resulting in a high market share of unqualified samples. The thin layer identification spots of the unqualified Arnebiae Radix sampled in 2015 were not consistent with the qualified samples. The thin layer identification of the unqualified samples sampled in 2022 was consistent with those of the qualified samples,but the depth of the spots were not consistent with those of the qualified samples,indicating that the current unqualified samples were adulterated samples,which posing greater challenges to the quality supervision of Arnebiae Radix. Through the exploratory research of twice National Drug Sampling and Inspection Project,it is preliminarily believed that it is of great significance to improve the standard test items of Arnebiae Radix,scientifically establish the limit value and strengthen the construction of the quality control system for the supervision.

  • Chao-jie LIAN, Sheng-yun DAI, Jie LIU, Li-nong GUO, Fei QIAO, Rui YANG, Hui-juan WANG, Jian ZHENG, Jie LIU
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 766-771.
    Objective:

    To determine the color of Arnebiae Radix, and the contents of six main purple pigment components (acetylshikonin,β-acetoxyisovalerylalkannin, deoxyshikonin, isobutylshikonin,β,β’-dimethylacrylalkannin and isovalerylshikonin) in Arnebiae Radix, and to study on the correlation between the color of Arnebiae Radix and the contents of six main purple pigment components.

    Methods:

    The La,and b values of the sample powder were determined using a spectrophotometer to characterize the color of Arnebiae Radix. The International Commission on Illumination (CIE) had developed a Lab color model,which was a digital description of human vision. A higher L value indicated greater brightness,a higher a value indicated redness and a lower a value indicates greenness,and a higher b value indicated yellowing and a lower b value indicates blueness. The contents of purple pigment components were determined using high performance liquid chromatography (HPLC),and the correlation between La,and b values and the contents of six main purple pigments was calculated using SPSS software.

    Results:

    The contents of acetylshikonin in 135 batches of samples ranged from 0.01% to 3.39%. The contents of β-acetoxyisovalerylalkannin ranged from 0.00% to 1.95%. The contents of deoxyshikonin ranged from 0.00% to 0.23%. The contents of isobutylshikonin ranged from 0.01% to 1.13%. And the contents of isovalerylshikonin ranged from 0.02% to 2.88%. The contents of β,β’-dimethylacrylalkannin ranged from 0.01% to 2.17%. There was a significant negative correlation between the contents of acetylshikonin,β-acetoxyisovalerylalkannin and isobutylshikonin and the L (black_white) chromaticity value of Arnebiae Radix,with a Spearman correlation coefficients between -0.138 and -0.222. The chromaticity value of a (red_green) was related to the five components other than acetylshikonin,which were β-acetoxyisovalerylalkannin,deoxyshikonin,isobutylshikonin,β,β’-dimethylacrylalkannin,and isovalerylshikonin,with a spearman correlation coefficients between 0.176 and 0.355;b (blue_yellow) chromaticity value was related to the five components other than β-acetoxyisovalerylalkannin,which were acetylshikonin,deoxyshikonin,and isobutylshikonin,β,β’-dimethylacrylalkannin. β,β’-dimethylacrylalkannin was positively correlated with a coefficient of 0.290,and negatively correlated with the other four components with a coefficients between -0.325 and -0.633.

    Conclusion:

    It is recommended that the assay limits of Arnebiae Radix be revised to β,β’-dimethylacrylalkannin not less than 0.30% and isovalerylshikonin not less than 0.29%.

  • Li-jun ZHANG, Peng LI, Long-hai LI, Xin-yue ZHANG, Dong-ni MA, Zhuan-ping ZHANG, Fang-di HU, Lei WANG, Hong-mei KE, Peng LI, Li LIU, Chun-yan WU, Xiao-an LI
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 903-911.
    Objective:

    To establish HPLC characteristic chromatogram and multi-index component determination methods,so as to effectively distinguish Pueraria lobata (Willd.) Ohwi (GH),Pueraria thomsonii Benth (FGH) and their confusing species,and to evaluate the quality of Puerariae Flos.

    Methods:

    HPLC was used to analyze the chemical components from different Puerariae Flos with different commercial specifications. The chromatographic conditions were optimized,and the characteristic chromatogram was established. Principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) analysis were used to analyze the characteristic chromatogram data of Puerariae Flos with different commercial specifications and to select the differential markers of Puerariae Flos. The differential markers were identified by comparing them with the reference substance,and their contents were determined and the determination results were analyzed.

    Results:

    HPLC characteristic chromatograms of GH,FGH and their confusing species were established,and a total of 18,27,18 and 8 common peaks were matched in GH,FGH,Pueraria lobata var. montana (GMM) and Wisteria sinensis (Sims) Sweet (ZTH) respectively. Ten chromatographic peaks were identified,including puerarin (Pu),daidzin (Da),daidzein (Dae),genistin (Ge),genistein (Gee),kakkalide (Ka),glycitin (Gl),tectoridin (Td),tectorigenin (Tg) and tectorigenin-7-O-xylosylglucoside (Tx),and HPLC method for the determination of the above ten components was established simultaneously. The results of PCA and PLS-DA could distinguish four kinds of samples with different commercial specifications. Gl,Tx,Ge,Td,Ka and one unknown compound 6 were the components with significant differences between the samples with four different commodity specifications. The chemical components of GH and FGH were obviously different from their adulterants. The content of Ka in GH were 10 times as much as that in FGH,and the content of Ge was half as much as that in FGH. Da,Gl was not detected in GH.

    Conclusion:

    The established characteristic chromatogram,chemometrics and multi-index component determination method in this study is rapid,simple,objectively and effectively and comprehensive quality evaluation of Puerariae Flos,which can provide the references for improving the quality control standard of Puerariae Flos.

  • Yong-qi ZHAO, Hong-wei ZHANG, Zhen-ling ZHANG, Ya-jing LI, Yi-ming WANG
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 893-902.
    Objective:

    To provide a basis for determining the quality markers(Q-Marker) of Lilii Bulbus decoction prepared slices by analyzing the Q-Marker of Lilii Bulbus by fingerprint,multi-component content determination and network pharmacology.

    Methods:

    The fingerprints of 15 batches of Lilii Bulbus prepared slices were established by HPLC,and the common peaks were analyzed chemometrically to screen out the differential components. The contents of the differential components were determined to compare the differences between samples from different habitats. The main pathways through which the differential components of Lilii Bulbus exert their antidepressant effects were analyzed based on network pharmacology,and the differential components were subjected to in vitro cellular antidepressant assays,and finally the Q-Markers of Lilii Bulbus prepared slices were analyzed based on the principle of quality marker screening.

    Results:

    Sixteen common peaks were identified in the 15 batches of Lilii Bulbus extracts,all with similarities above 0.931,7 of which were identified and 4 differential components were screened using chemometrics. The content of the components of regaloside A,regaloside B,and regaloside C were determined in the 15 batches of Lilii Bulbus extracts,and the network pharmacological study revealed that the differential components might exert their antidepressant effects through the targets of TNF,GAPDH and MAPK3,regulating. The results of cellular experiments showed that compared with the model group,the cellular viability of the low,medium and high dose groups of Lilii Bulbus extract was significantly increased (P<0.01),and the cellular viability of the low,medium and high dose groups of regaloside A,regaloside B,and regaloside C was significantly increased (P<0.01).

    Conclusion:

    The fingerprinting and network pharmacology studies suggested that regaloside A,regaloside B,and regaloside C as quality markers of Lilii Bulbus prepared slices,which can provide reference for Lilii Bulbus quality control and pharmacological efficacy studies.

  • Mei-mei GAO, Jian-you HUANG, Ya-nan ZHAI, Ling-feng JIANG, Guo-shou LU, Xiao-xi HU, Xue-qing XIA, Dong-mei LI
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 882-892.
    Objective:

    To base on the method of HPLC fingerprint,multi-component quantification and chemical pattern recognition,to evaluate the quality of leaves of Cunninghamia lanceolata from different producing areas and to provide basis for further development and utilization.

    Methods:

    High-performance liquid chromatography (HPLC) was used to determine the contents of amentoflavone,bilobetin,hinokiflavone,ginkgetin,isoginkgetin and sciadopitysin in the Cunninghamia lanceolata. Fingerprints of 10 batches of Cunninghamia lanceolata from different habitats were established. Based on the common peak area of the fingerprint,the overall quality of Cunninghamia lanceolata was evaluated by principal component analysis(PCA),orthogonal partial least squares discriminant analysis(OPLS-DA),statistical analysis,and pattern recognition chemometrics methods.

    Results:

    A total of 14 common peaks in 10 batches of leaves of Cunninghamia lanceolata,and the similarity ranged from 0.955 to 1.000 with good consistency. The mass fractions of six biflavones in the sample were 2.42-5.24 mg·g-1,0.10-0.24 mg·g-1,1.55-3.67 mg·g-1,0.21-0.89 mg·g-1,0.10-0.24 mg·g-1 and 0.51-2.39 mg·g-1,respectively,including amentoflavone,bilobetin,hinokiflavone,ginkgetin,isoginkgetin and sciadopitysin. According PCA,the difference in the quality of Cunninghamia lanceolata from different habitats was further evaluated. Ten batches of medicinal materials were divided into three major categories,and four main factors affecting the classification of Cunninghamia lanceolata were found. Finally,OPLS-DA screened the excellent spectral peaks 6,12,7 (7-demethylated ginkgo biloba biflavone),13 (kumatsu biflavone),5 (Amentotaxus argotaenia biflavone),9 (chamaecypress biflavone) and 2,etc,seven differential markers can be used to distinguish different batches of Cunninghamia lanceolata.

    Conclusion:

    The established method is simple to stable and reliable. Combined with chemical pattern recognition,it can be used for the quality evaluation of Cunninghamia lanceolata.

  • Meng-qi LU, Xue-jing ZHANG, Meng-dan XU, Xiao-kang LIU, Ji-yu GONG, Tian-zhu ZHANG
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 874-881.
    Objective:

    To explore the effects of key operating units such as different water contents and drying methods in processing methods on the content of main active ingredients in Atractylodes chinensis(DC.)Koidz. fresh cutting slices. To develop an HPLC method for simultaneous determination of five active ingredient (atractylenolide Ⅲ,atractylenolideⅠ,atractylodin,β-eudesmol,atractylon) in Atractylodes chinensis.

    Methods:

    The Supersil ODS2 column (250 mm×4.6 mm,5 μm ) was used,the mobile phase was 0.1% phosphoric acid (A)- methanol(B) with gradient elution,at the flow rate of 1.0 mL·min-1,the detection wavelength was 220,270,203 nm,and the column temperature was 30 ℃. The entropy weight-TOPSIS method and cluster analysis were combined to determine the best method of fresh cutting process of Atractylodes chinensis.

    Results:

    The validation results of the content determination methodology were good. The contents of five components(atractylenolide Ⅲ,atractylenolide Ⅰ,atractylodin,β-eudesmol,atractylon) were 0.052-0.243,0.195-1.015,2.022-4.418,0.119-5.049,0.209-8.638 mg·g-1,respectively. The optimal cutting moisture content range of Atractylodes chinensis fresh cutting was (40%±3%)-(50%±3%). The results of ANOVA showed that the contents of atractylenolide Ⅲ,atractylenin,β-eudesmol and atractylone were significantly higher in the seven processing(S1-S7) of fresh cutting than in the traditional raw-tanning group(S8). The entropy weight-TOPSIS analysis showed that 60 ℃ microwave drying,40 ℃ microwave drying and 50 ℃ blast drying slices pieces have better quality. Cluster analysis resulted in three groups,accounting for 12.5%,50% and 37.5% respectively.

    Conclusion:

    The results indicate that different drying methods and temperatures have a significant impact on the quality of Atractylodes chinensis slices. This study is valuable for the subsequent research in Atractylodes chinensis fresh cutting slices.

  • Zhen-xia ZHAO, Yun GENG, Rong LEI, -Xuan YIN, Yong-li LIU
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 796-805.
    Objective:

    To establish a method for simultaneous determination of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,cynarin,galuteolin,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C in Shanjujiangya capsules by HPLC and analyze compositional change of Chrysanthemi Flos combined with law of quantity transfer.

    Methods:

    The analysis was performed on Waters Symmetry C18 column (250 mm×4.6 mm,5 μm),with mobile phase composed of acetonitrile -0.1% phosphoric acid solution at a flow rate of 1.0 mL·min-1 in gradient elution mode. The column temperature was 30 ℃ and the detection wavelength was 328 nm. The transfer rates of the above eight components were used as the indexes for quality evaluation to study the quantity value of transfer rule from the decoction piece to the extracting solution.

    Results:

    The results showed that the determination of eight components manifested a good linear relationship in the range of mass concentration (r>0.999 9),the average recoveries of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,cynarin,galuteolin,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C were 98.3%-101.9%,with RSDs of 0.066%-0.64%. The contents of the above 8 components in 3 samples were 0.257-0.279 mg·g-1,0.629-0.650 mg·g-1,0.402-0.476 mg·g-1,0.454-0.539 mg·g-1,1.118-1.278 mg·g-1,0.653-0.740 mg·g-1,0.659-0.706,1.138-1.167 mg·g-1,respectively.

    Conclusion:

    The HPLC method established in this study is simple,repeatable and stable. The analysis of quantity transfer provides data support for the establishment of content methods and the formulation of limits. This study can provide basis for quality control method of Shanjujiangya capsules.

  • Jun-sheng HAO, Hong-ying BAO, Si-ri-gu-leng, He MENG, Ya-ting REN, Yan HONG, Ying XIN
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 816-826.
    Objective:

    To establish HPLC fingerprint and determine caffeic acid,isoquercitrin,quercitrin,rosmarinic acid,lithospermic acid and salvianolic acid B in Tournefortia sibirica Linnaeus,and to provide evidence for quality control of Tournefortia sibirica Linnaeus.

    Methods:

    The chromatographic separation was performed on an Agilent Eclipse Plus C18 column (250 mm×4.6 mm,5 μm) with gradient elution (0-60 min,9%B→38%B) of 0.2% phosphoric acid(A) and acetonitrile(B) . The detection wavelength was 330 nm. The column temperature was kept at 40 ℃ and the flow rate was 1.0 mL·min-1. The HPLC fingerprint of all batches of Tournefortia sibirica Linnaeus was established using Similarity Evaluation Software for Chromatographic Fingerprint of Traditional Chinese Medicine (2012 edition) and the common peaks were identified by reference standards. Six constituents in Tournefortia sibirica Linnaeus were quantified.

    Results:

    Eleven common peaks were confirmed,and 6 common peaks were identified by reference standards including caffeic acid,isoquercetin,quercetin,rosmarinic acid,purple oxalic acid,and salvianolic acid B. The similarities of 17 batches samples were 0.931 to 0.996. By the methodology examination,RSDs for the precision test were 1.9%,1.0%,1.4%,0.19%,1.1% and 0.32%,respectively. RSDs for the reproducible test were 3.5%,2.3%,3.3%,0.14%,1.1% and 0.19%,respectively. RSDs for the stability test were 0.80%,1.1%,1.7%,0.52%,0.54% and 0.78%,respectively. Caffeic acid,isoquercitrin,quercitrin,rosmarinic acid,lithospermic acid and salvianolic acid B had good separation and showed good linearity in their respective linear ranges. The average recoveries ranged from 95% to 105%,and the contents (calculated with reference to the dried drug) were between 0.004% to 0.013%,0.030% to 0.259%,0.032% to 0.256%,0.256% to 1.246%,0.018% to 0.072% and 0.062% 0.499%,respectively.

    Conclusion:

    The established HPLC fingerprint and quantification method is stable and reliable,which can provide basis for the quality control of Tournefortia sibirica Linnaeus.

  • Lu-di JIANG, Xiao-yan WANG, Ya-nan LIU, Hai-bo WANG, Yan YANG, Hai-yan LI
    Chinese Journal of Pharmaceutical Analysis. 2024, 44(5): 827-834.
    Objective:

    To establish an HPLC-MS/MS method for simultaneous determination of 11 components(harpagide,salidroside,nuezhenide,lobetyolin,wedelolactone,harpagoside,vaccarin,6-gingerol,atractylenolide Ⅲ,atractylenolide Ⅱ,and atractylenolide Ⅰ)in Gengnianning.

    Methods:

    HPLC assay was performed on C18 column(100 mm×2.1 mm,1.9 μm)with a mixture of methanol and 0.1% formic acid as the mobile phase in gradient elution at a flow rate of 0.3 mL·min-1. The column temperature was 25 ℃ and the injection volume was 1 μL. Detection was carried out on a triple quadrupole mass spectrometer in positive ion mode(harpagide,salidroside,nuezhenide,lobetyolin,wedelolactone and harpagoside)and negative ion mode(vaccarin,6-gingerol,atractylenolide Ⅲ,atractylenolide Ⅱ,and atractylenolide Ⅰ)using an electrospray ion source(ESI). Multiple reaction monitoring(MRM)mode was employed.

    Results:

    The calibration curves were linear within the ranges of 1.485-29.71 μg·mL-1,1.620-32.40 μg·mL-1,7.801-156.0 μg·mL-1,0.518-10.35 μg·mL-1,0.167-3.333 μg·mL-1,0.359-7.179 μg·mL-1,1.455-29.10 μg·mL-1,1.520-30.40 μg·mL-1,0.160-3.205 μg·mL-1,0.143-2.864 μg·mL-1 and 0.157-3.136 μg·mL-1 for harpagide,salidroside,nuezhenide,lobetyolin,wedelolactone,harpagoside,vaccarin,6-gingerol,atractylenolide Ⅲ,atractylenolide Ⅱ,and atractylenolide Ⅰ,respectively. All 11 components showed good linearity(r≥0.998 0). The average recoveries(n=6)were in the range of 95.9%-102.6% with RSDs within 0.90%-3.0%. The contents of harpagide,salidroside,nuezhenide,lobetyolin,wedelolactone,harpagoside,vaccarin,6-gingerol,atractylenolide Ⅲ,atractylenolide Ⅱ and atractylenolide Ⅰ in 10 tested samples from 5 manufactures were 14.8-104.5,37.6-288.5,335.8-1 332.8,6.2-10.1,6.6-61.8,13.7-75.1,57.4-132.8,16.9-70.6,11.8-33.9,3.4-15.4 and 6.5-12.9 μg·g-1.

    Conclusion:

    The developed method is accurate and sensitive. It can be used in quality control of Gengnianning.