目的: 采用多成分含量测定结合化学计量学方法比较克癀发酵前后的化学成分变化。方法: 采用ZORBAX Eclipse Plus C18（4.6 mm×250 mm,5 μm）色谱柱,以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱,流速1.0 mL · min-1,柱温30 ℃,检测波长203 nm,进样量10 μL。对克癀发酵前后所含的绿原酸、升麻素苷、三七皂苷R1、黄芩苷、小檗碱、槲皮素、黄芩素、汉黄芩素、大黄素、大黄酚和大黄素甲醚11个化学成分进行含量测定;建立克癀发酵前后的化学指纹图谱;应用聚类分析（HCA）、主成分分析（PCA）及正交偏最小二乘判别分析（OPLS-DA）等化学计量学方法,对克癀发酵前后成分进行区分与比较,筛选主要差异性成分。结果: 建立了克癀发酵前后的化学指纹图谱,共标定了22个共有峰,所建立的化学指纹图谱专属性良好,可用于克癀的质量评价;多成分含量测定和化学计量学结果表明,克癀原药材粉末、克癀发酵粉末和克癀胶囊内容物粉末中三七皂苷R1（1.023~2.927 mg · g-1）、黄芩素（0.125~0.568 mg · g-1）和盐酸小檗碱（2.151~3.068 mg · g-1）的含量有明显差异;确认三七皂苷R1、黄芩素和小檗碱为克癀发酵前后的差异性成分,可以作为区分和鉴别克癀发酵前后的质量标志物。结论: 经过发酵的克癀化学成分发生了明显变化,明确了其差异性成分,为克癀胶囊生产质量控制提供了分析依据。
王玉霞
,
侯佳豪
,
李冉
,
涂天智
,
宋永兴
,
段绪红
,
马东来
,
王茜
. 基于多成分含量测定和化学计量学的克癀胶囊发酵前后成分比较研究*[J]. 药物分析杂志, 2025
, 45(5)
: 833
-842
.
DOI: 10.16155/j.0254-1793.2024-1143
Objective: To compare the chemical composition changes in Kehuang before and after fermentation using multi-component content determination combined with chemometric analysis. Methods: HPLC was employed using a ZORBAX Eclipse Plus C18 column (4.6 mm×250 mm, 5 μm). The mobile phase consisted of acetonitrile and 0.1% phosphoric acid aqueous solution, applied with gradient elution. The flow rate was set at 1.0 mL · min-1,the column temperature was maintained at 30 ℃, the detection wavelength was 203 nm, and injection volume was 10 μL. This method was applied to analyze the chemical compositions of Kehuang before and after fermentation and to establish their corresponding chemical fingerprint profiles. Quantitative determination was conducted for 11 chemical constituents, including chlorogenic acid, cimifugin, notoginsenoside R1, baicalin, berberine, quercetin, baicalein, wogonin, emodin, chrysophanol, and physcion. Furthermore, chemometric methods such as hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were employed to distinguish and compare the samples before and after fermentation, with the aim of identifying key differential compounds. Results: Chemical fingerprint profiles were established for Kehuang before and after fermentation, with 22 common peaks identified. The fingerprints had good specificity and can be used for quality evaluation of Kehuang. The results of multi-component content determination and chemometrics analysis revealed significant differences in the content of notoginsenoside R1 (1.023-2.927 mg · g-1), scutellarein (0.125-0.568 mg · g-1), and berberine hydrochloride (2.151-3.068 mg · g-1) among the crude Kehuang powder, fermented Kehuang powder, and the content of Kehuang capsules. Notoginsenoside R1, scutellarein and berberine were identified as differential markers between pre- and post-fermentation of Kehuang, which could serve as quality indicators for distinguishing and identifying Kehuang before and after fermentation. Conclusion: The chemical composition of Kehuang changes significantly after fermentation, and the differential compounds have been clearly identified, which provides an analytical basis for quality control in the production of Kehuangcapsules.
[1] LI L,WANG L,FAN W,et al.The application of fermentation technology in traditional Chinese medicine:a review[J].Am J Chin Med,2020,48(4):899
[2] YANG HY,HAN L,LIN YQ,et al.Probiotic fermentation of herbal medicine:progress,challenges,and opportunities[J].Am J Chin Med,2023,51(5):1105
[3] 付婷婷,尹兴斌,蔡梦如,等.曲类中药的品种整理和标准现状分析[J].中国中药杂志,2023,48(10):2699
FU TT,YIN XB,CAI MR,et al.Varieties systematization and standards status analysis of fermented Chinese medicine[J].China J Chin Mater Med,2023,48(10):2699
[4] 倪沁雨,林佳成,涂天智,等.基于网络药理学探讨克癀胶囊抗肝纤维化的作用机制[J].中西医结合肝病杂志,2023,33(6):518
NI QY,LIN JC,TU TZ,et al.The mechanism of Kehuang capsule against hepatic fibrosis based on network pharmacology[J].Chin J Integr Tradit West Med Liver Dis,2023,33(6):518
[5] 周艾强,关卫兵,王凤林,等.克癀胶囊对非酒精性脂肪肝大鼠的影响[J].中成药,2019,41(8):1969
ZHOU AQ,GUAN WB,WANG FL,et al.Effects of Kehuang capsule on rats with nonalcoholic fatty liver[J].Chin Tradit Pat Med,2019,41(8):1969
[6] 纪翠芳,章明敏.克癀胶囊改用人工麝香、人工牛黄前后对肝癌细胞的影响[J].中国现代药物应用,2020,14(13):249
JI CF,ZHANG MM.Effect of Kehuang capsule on hepatocellular carcinoma cells before and after using artificial musk and artificial bezoar[J].Chin J Mod Drug Appl,2020,14(13):249
[7] 刘逆夫,娄勇军.HPLC法测定克癀胶囊中人参皂苷Rg1的含量[J].中国中医药现代远程教育,2007,5(7):26
LIU NF,LOU YJ.Determination of ginsenoside Rg1 in Kehuang capsule by HPLC[J].Chin Med Mod Dis Educ China,2007,5(7):26
[8] 叶曼红. 液相色谱法快速测定克癀胶囊中3种皂苷含量[J].海峡药学,2009,21(8):58
YE MH.Content determination of 3 gensenosides in Kehuang capsule by ultra performance LC[J].Strait Pharm J,2009,21(8):58
[9] 吴聪泉,马鸿杰,刘冬凤.克癀胶囊中麝香酮组分的含量检测研究[J].世界最新医学信息文摘,2017,17(86):199
WU CQ,MA HJ,LIU DF.Study on the content detection of muscone components in Kehuang capsule[J].World Latest Med Inf,2017,17(86):199
[10] 李晓静,刘雅茹,李冉,等.克癀胶囊治疗非酒精性脂肪性肝炎小鼠药效学研究[J].中西医结合肝病杂志,2024,34(3):230
LI XJ,LIU YR,LI R,et al.Pharmacodynamic study of Kehuang capsule in mice with nonalcoholic steatohepatitis[J].Chin J Integr Tradit West Med Liver Dis,2024,34(3):230
[11] 胡超,贺红,杨岩涛,等.片仔癀HPLC指纹图谱的总量统计矩及其相似度法分析[J].中国实验方剂学杂志,2016,22(8):53
HU C,HE H,YANG YT,et al.HPLC fingerprint of Pien Tze Huang by total statistical moment and similarity method[J].Chin J Exp Tradit Med Form,2016,22(8):53
[12] TANG L,SHAO J,ZHENG H,et al.Cleaner production of baicalein by novel high-temperature-little-acid hydrolysis of baicalin[J].Green Chem Lett Rev,2024,17(1):2319698
[13] YIM JS,KIM YS,MOON SK,et al.Metabolic activities of ginsenoside Rb1,baicalin,glycyrrhizin and geniposide to their bioactive compounds by human intestinal microflora[J].Biol Pharm Bull,2004,27(10):1580
[14] GUO FY,LI CH,DOU JX,et al.Research progress on pharmacological properties and application of probiotics in the fermentation of Scutellaria baicalensis Georgi[J].Front Nutr,2024,11:1407182
