茯苓皮与白茯苓中三萜类成分的HPLC指纹图谱与化学模式识别研究.pdf

天然产物研究与开发NatProdResDev2023,35:1469-1479茯苓皮与白茯苓中三枯类成分的HPLC指纹图谱与化学模式识别研究占慧慧1.2.3，丁婵1.2 3，彭思源1.2.3,刘媛1-2 3，孟俊华1.2.3，肖作为1*，崔培梧1.2.3*湖南中医药大学药学院；国家中医药管理局中药药性与药效三级科研实验室；3湖南中医药大学菌物药研究室，长沙410 2 0 8摘要：建立茯苓皮与白茯苓中三类成分的HPLC 指纹图谱,并对其进行化学模式识别研究,为菌物药茯苓的质量控制及标准制定提供依据。采用HPLC法,Agilent5TC-Cis（2）色谱柱（2 50 mm4.6mm,5m）,以乙腈-0.3%磷酸水溶液为流动相梯度洗脱，流速1.0 mL/min，柱温2 5,变波长方式检测，采集波长为2 42、2 0 3nm，进样量10 L。建立了茯苓皮与白茯苓中三类成分指纹图谱,应用聚类分析（CA）、主成分分析（PCA)手段结合相似度评价对实验数据进行识别,以分析不同茯苓样品的相似性和差异性，找出可反映茯苓皮与白茯苓三成分特征的关键色谱峰。茯苓皮与白茯苓三成分指纹图谱的共有模式下确认了6 个共有峰，分别为2 号峰（茯苓新酸B）、3号峰（去氢土莫酸）、4号峰（茯苓新酸A）9号峰（去氢茯苓酸）、10 号峰（茯苓酸）、12 号峰（松苓新酸）；不同来源的茯苓样品相同药用部位间相似度均在0.90 以上；依据CA和PCA分析结果将茯苓样品药用部位分为两大类。PCA分析结果显示包括代表去氢土莫酸、去氢茯苓酸、茯苓酸成分在内的7 个共有色谱峰（3、5和6、8 11号峰）在评价茯苓皮及白茯苓质量时必不可少,其余包括代表茯苓新酸B、茯苓新酸A、松苓新酸成分在内的8 个色谱峰（1和2、4、7、12 15号峰)可作为评价茯苓皮质量的特征鉴别峰;白茯苓中15个共有色谱峰间的差异比茯苓皮小。本研究建立的茯苓三HPLC指纹图谱和化学模式识别可为不同茯苓药材质量控制与评价提供参考。关键词：茯苓；三；指纹图谱；聚类分析；主成分分析；质量控制中图分类号：R284.1D0I:10.16333/j.1001-6880.2023.9.001Study on HPLC fingerprint and chemical pattern recognitionof triterpenoids in Poriae Cutis and White PoriaZHAN Hui-hil-23,DING Chan-2,PENG S-yuanLIU Yuan23,MENG Jun-hual-2.3,XIAO Zuo-weil*,CUI Pei-wuCollege of Pharmacy,Hunan University of Chinese Medicine;2 Research Lab of TCM Property&Efficacy,Tier 3,National Administration of TCM;3 Mycomedicine Research Lab,Hunan University of Chinese Medicine,Changsha 410208,ChinaAbstract:In this study,HPLC analysis combined with chemical pattern recognition was employed to establish the HPLC fin-gerprint related to triterpenoids in Poriae Cutis and White Poria,which can provide a reference for quality control and standardestablishment of decoction pieces derived from mycomedicine Poria cocos.HPLC method was adopted and performed on anAgilent 5 TC-Cis(2)column(250 mm 4.6 mm,5 m).The mobile phase was consisted of acetonitrile and 0.3%phos-phoric acid solution,and gradient elution procedure was employed with the flow rate setting at 1.O mL/min.The column tem-perature was set at 25 C,the detection wavelength was set at 242 and 203 nm,and the injection volume was 10 L,respec-tively.To establish the HPLC fingerprint and chemical pattern recognition of Poriae Cutis and White Poria,cluster analysis收稿日期:2 0 2 3-0 2-2 0基金项目：国家自然科学基金（8 197 32 11/C0033375）；湖南省中医药科研计划（D2022139（A 2 0 2 2 0 0 5-17）；湖南省大学生创新创业训练计划（湘教通2 0 2 2 17 4号-2 918）；湖南中医药大学“十四五”重点学科-生物工程学科（校行发规字2 0 2 3 2 号）*通信作者E-mail:,文献标识码：A接受日期：2 0 2 3-0 7-0 5文章编号：10 0 1-6 8 8 0(2 0 2 3)9-146 9-11,1,2,31,1,2,3*1470(CA),principal component analysis(PCA)and similarity evaluation were adopted to process the experimental data,the sim-ilarity and difference of Poriae Cutis and White Poria samples were analyzed subsequently.Finally,the key chromatographicpeaks representing triterpenoids were pointed out from the established HPLC fingerprint.Among the common chemical compo-nents showed in the chromatograms,six peaks related to poricoic acid B(peak 2),dehydrotumulosic acid(peak 3),poricoicacid A(peak 4),dehydropachymic acid(peak 9),pachymic acid(peak 10),and dehydrotrametenolic acid(peak 10)wereindentified,respectively.The similarity of tested decoction pieces from the same part of P.cocos sclerotium were all above O.90.According to CA and PCA data,all the decoction pieces analyzed in this study can be divided into two groups representingdifferent medicinal parts of P.cocos sclerotium.Seven common chromatographic peaks including peak 3(dehydrotumulosicacid),peak 5,peak 6,peak 8,peak 9(dehydropachymic acid),peak 10(pachymic acid)and peak 11 were essential for thequality evaluation of Poriae Cutis and White Poria,the other eight chromatographic peaks including peak 1,peak 2(poricoicacid B),peak 4(poricoic acid A),peak 7,peak 12(dehydrotrametenolic acid)and peaks 13 to 15 could be used as thecharacteristic identification peaks for evaluating the quality of Poriae Cutis according to the loading scatter analysis of PCAdata.The characteristic difference deduced from the 15 common chromatographic peaks of White Poria samples was smallerthan that of Poriae Cutis samples.The established HPLC fingerprint of Poriae Cutis and White Poria combined with chemical patternrecognition can provide a scientific reference for the quality control and evaluation of samples or products derived from P.cocos.Key words:Poria cocos;triterpenoids;fingerprint;cluster analysis;PCA;quality control茯苓又名茯、茯灵、松等，系多孔菌科真菌茯苓Poria cocos（Sc h w.）Wo l f 的干燥菌核，味甘、淡,性平,归心、肺、脾、肾经,有利水渗湿、健脾宁心的功效 。茯苓收载于中华人民共和国药典（简称中国药典）2 0 2 0 年版一部,是中药四君八珍之一,有“十方九苓 之称2 。茯苓的主要药效物质是茯苓多糖和茯苓三枯3,现代药理学研究表明，茯苓三具有利尿4、抗肿瘤5、免疫调节6 、抗炎7 、抗衰老8 、降血糖和调血脂9 等作用,是当前用于表征茯苓质量和效应的重要指标10-2 。目前市场流通的茯苓主要为栽培品种，不同的产地、加工方式、饮片规格等都可能影响茯苓药材和饮片的质量1.1。2 0 2 0 年版中国药典仅采用薄层色谱法鉴别控制茯苓和茯苓皮药材的质量，难以全面反映其质量的优劣。指纹图谱研究是现代中药领域内普遍被认可、应用较广泛的中药质量控制方法之一,具有整体性高、特征性强等优势,符合中药多成分综合作用的特点，有进一步鉴别、鉴定和分类的潜力14,15。化学模式识别可对指纹图谱的多维信息进行分析,准确反映药材的质量差异,从而达到整体描述和合理评价药材质量的目的13。本研拟究建立茯苓皮（源于茯苓菌核表皮）与白茯苓（源于茯苓菌核去皮后的部位)的HPLC指纹图谱,并结合模式识别手段分析各共有峰对指纹图谱的作用，找出其中的主要峰和特征峰,从整体上评价茯苓皮和白茯苓中各化学物质的差异，为茯苓药材的质量控制提供依据。天然产物研究与开发1仪器与试药1.1仪器Agilent1200高效液相色谱仪（美国安捷伦科技公司）;JS-40型超声波清洗仪（常州鸿泽实验科技有限公司）。1.2试药对照品茯苓新酸B（批号140 7 2 3，质量分数98%）、茯苓新酸A（批号140 314，质量分数98%）、去氢茯苓酸（批号140 90 3，质量分数98%）、松苓新酸（批号140 90 2，质量分数98%）、茯苓酸（批号140 2 12，质量分数98%）购自成都克洛玛生物科技有限公司；去氢土莫酸（批号AF21032,质量分数98%）购自成都埃法生物科技有限公司;乙腈为色谱纯,水为怡宝纯净水,其余试剂为分析纯。茯苓药材样品来源见表1,经湖南中医药大学刘塔斯教授鉴定为多孔菌科真菌茯苓Poria cocos（Sc h w.）Wo l f 的不同药用部位，符合中国药典2020年版各项规定。2方法与结果2.1色谱条件Agilent 5 TC-Cis（2)（2 50 m m 4.6 m m,5m)色谱柱;流动相为0.3%磷酸水溶液（A）-乙腈(B),梯度洗脱:0 18 min,50%60%B;1828min,60%B;28 58 min,60%92%B;58 68 min,92%B;检测波长为2 42 nm和2 0 3nm;进样量10L;柱温2 5;体积流量1.0 mL/min。Vol.35Vol.35编号No.S1S2S3S4S5S6S7S8S9S10S11S12S13S14占慧慧等：茯苓皮与白茯苓中三枯类成分的HPLC指纹图谱与化学模式识别研究药用部位Medicinal part茯苓皮PoriaeCutis茯苓皮PoriaeCutis茯苓皮PoriaeCutis茯苓皮Poriae Cutis茯苓皮PoriaeCutis茯苓皮PoriaeCutis茯苓皮PoriaeCutis茯苓皮PoriaeCutis茯苓皮PoriaeCutis白茯苓WhitePoria白茯苓WhitePoria白茯苓WhitePoria白茯苓WhitePoria白茯苓WhitePoria1471表1样品信息Table 1 Information of samples生产企业Manufacturer安徽三义堂中药饮片有限公司亳州市中药饮片厂云南鸿翔中药科技有限公司湖南振兴中药有限公司康美药业股份有限公司四川藏曦堂生物科技有限公司亳州市泽信堂药业有限公司河北纽恩堂电子商务有限公司湖南聚仁中药饮片有限公司湖南聚仁中药饮片有限公司康美药业股份有限公司亳州市泽信堂药业有限公司云南国鹤药业有限公司岳阳天香中药饮片有限公司产地Origin云南安徽云南湖南湖南四川云南云南云南湖南湖南云南云南云南批号Lot No.2103009202110252021102321073109200801781202111092021110320211112210113012108310121031501120211031521070082021111072.2溶液的制备2.2.1混合对照品溶液的制备精密称取各茯苓三酸对照品适量,加甲醇溶解制成茯苓新酸B、去氢土莫酸、茯苓新酸A、去氢茯苓酸、茯苓酸、松苓新酸对照品质量浓度分别为0.020 7,0.071 4,0.006 4,0.038 6,0.037 1,0.017 1 mg/mL的混合对照品溶液。放于4冰箱保存,备用。2.2.2供试品溶液的制备取茯苓样品粉末2.0 g（过6 0 目筛）,精密称定,置10 0 mL干燥具塞锥形瓶中,加甲醇50 mL,密塞,称定质量,超声处理30 min，放冷,用甲醇补足减失质量，滤过，茯苓皮样品滤液经0.2 2 m微孔滤膜过滤,取续滤液，即得；白茯苓样品滤液先旋转蒸干后定容至2.0 mL,再经0.2 2 m微孔滤膜过滤，取续滤液，即得。2.3方法学考察2.3.1精密度实验取云南茯苓皮样品（S8），按“2.2.2”项下方法制备供试品溶液，依据“2.1”项下的色谱条件，连续进样测定6 次,以茯苓酸（10 号峰为参照峰,计算得各色谱峰的相对保留时间和相对峰面积的RSD值分别在0.0 4%0.8 6%和0.2 2%1.6 9%之间，表明仪器精密度良好。2.3.2稳定性实验取云南茯苓皮样品（S8），按“2.2.2”项下方法制备供试品溶液，依据“2.1”项下的色谱条件，分别于0、2、4、6、8、12、2 4h进样测定,测得各色谱峰的相对保留时间和相对峰面积并计算得RSD值分别在0.0 1%0.2 7%和0.32%1.19%之间，表明供试品溶液在2 4h内稳定。2.3.3重复性实验取云南茯苓皮样品（S8）6 份，精密称定，按“2.2.2”项下方法制备供试品溶液，依据“2.1”项下的色谱条件进样测定，测得各色谱峰的相对保留时间和相对峰面积并计算得RSD值分别在0.0 2%0.29%和0.8 1%4.53%之间，表明方法重复性好。2.3.4溶剂峰和滞后峰的考察取空白对照（甲醇）和云南茯苓皮样品（S8）溶液,检测时间延长至2 h,考察结果如图1所示,测试结果表明空白溶剂无干扰且未见滞后峰出现。2.4#指纹图谱的建立2.4.1#指纹图谱共有模式的建立将14批不同产地不同药用部位茯苓样品的色谱图导人“中药色谱指纹图谱相似度评价系统（2 0 12 版）”软件分别生成茯苓样品的叠加图谱和对照指纹图谱。14批茯苓皮和白茯苓样品的叠加图谱和对照指纹图谱见图2,茯苓皮和白茯苓各自的对照指纹图谱及混合对照品HPLC色谱图见图3。由图2 和3可知，茯苓皮与白茯苓样品共有的典型色谱峰有15个；经与混合对照品溶液色谱图比对，1472天然产物研究与开发AVol.35B0.00Fig.1 Investigation chromatograms of solvent peaks(A)and hysteresis peaks(B)0图2 14批茯苓皮与白茯苓饮片的指纹图谱叠加图和对照指纹图谱Fig.2 Superimposed fingerprints and control fingerprint of 14 batches of Poriae Cutis and White Poria decoction pieces20.00图1溶剂峰(A)和滞后峰(B)的考察色谱图41298102040.0030时间Time(min)60.00时间Time(min)4080.0010,1150100.0013141560120.00RS14S13S12S11S10S9S8S7S6S5S4S3S2S14A121310(S)1415BC0图3不同产地茯苓皮(A)和白茯苓(B)饮片对照指纹图谱与混合对照品HPLC图(C)Fig.3Control fingerprints of Poriae Cutis（A)a n d Wh i t e Po r i a （B)d e c o c t i o n p i e c e s f r o mdifferent regions and HPLC chromatogram of mixed standards(C)注：2：茯苓新酸B;3：去氢土莫酸;4：茯苓新酸A;9：去氢茯苓酸;10：茯苓酸;12：松苓新酸。Note:2：Po r i c o i c a c i d B;3：D e h y d r o t u m u l o s i c a c i d；4:Poricoic acid A;9:Dehydropachymic acid;10:Pachymic acid;12:Dehydrotrametenolic acid.102030时间Time(min)405060Vol.35指认了其中6 个共有峰，分别为茯苓新酸B（2 号峰）、去氢土莫酸（3号峰）、茯苓新酸A（4号峰）、去氢茯苓酸9号峰）茯苓酸（10 号峰）、松苓新酸（12号峰);其中10 号峰茯苓酸成分是茯苓中具有代表性的活性三酸类成分，性质稳定,峰面积所占比例较大,分离度好,因此确定其为参照峰(S)。2.4.2茯苓皮与白茯苓共有峰的相对保留时间与相对峰面积以不同产地茯苓皮和白茯苓样品HPLC色谱图表2 14批茯苓皮与白茯苓饮片指纹图谱共有峰的相对保留时间Table 2Common peaks relative retention time of the fingerprints of 14 batches of Poriae Cutis and White Poria decoction pieces样品Sample1S10.4000.445 0.4920.5420.6680.7260.8770.922 0.9591.0001.0701.1121.158 1.185S20.3990.444 0.4920.5420.6690.727 0.8780.9220.959 1.0001.0691.112 1.156 1.184S30.3990.4420.4900.5380.6640.7240.8770.9220.9591.000 1.069 1.1121.1571.185S40.3980.4420.4890.5380.6640.7230.8760.9210.9581.000 1.0701.1131.1581.187S50.3980.4420.4890.5380.6650.7230.8770.9220.959 1.000 1.0701.113 1.1581.186S60.3980.4420.489 0.5380.6650.7240.8770.9220.9591.000 1.070 1.112 1.1581.185S70.3980.4420.4890.5380.6650.7240.8770.922 0.9591.000 1.0701.113S80.3950.4390.4870.5360.6630.7220.876 0.921 0.9591.0001.070 1.1131.1581.186S90.402 0.4460.4940.5440.6700.7280.8780.922 0.9591.000 1.0701.112 1.1571.1841.226S100.4040.4460.4960.5420.6670.728 0.8780.9220.9591.0001.0661.1081.1531.180S110.4030.4450.495 0.5410.6660.727 0.8770.922 0.9591.000 1.0661.1081.1531.180S120.4020.444 0.4940.5400.6660.7260.8770.9220.9591.000 1.068 1.1101.1551.182S130.403 0.4460.4950.5410.6670.7270.8780.922 0.9591.000 1.0661.109 1.1531.1801.222S140.403 0.445 0.4950.5410.6670.7270.8780.9220.959 1.0001.067 1.109 1.1531.181平均值0.4000.4440.4920.5400.6660.7250.8770.9220.9591.0001.0691.1111.1561.184Mean占慧慧等：茯苓皮与白茯苓中三类成分的HPLC指纹图谱与化学模式识别研究231473中的10 号峰(茯苓酸)为参照峰（S）,计算各个批次样品中其他共有峰与S峰的相对保留时间与相对峰面积,结果见表2 和表3。由表2 和表3可知,茯苓皮与白茯苓样品共有峰相对保留时间的波动较小（R SD 值在0.0 2%0.6 4%），而相对峰面积波动较大,表明茯苓皮与白茯苓HPLC 指纹图谱中主要峰群的整体面貌基本一致,但同一批次样品各成分之间含有量及不同药用部位样品各成分含有量差别较大。共有峰相对保留时间Relative retention time common peaks456789101112131.1581.18514151.2271.2261.2271.2301.2281.2271.2271.2281.2211.2211.2241.2221.225表314批茯苓皮与白茯苓饮片指纹图谱共有峰的相对峰面积共有峰相对峰面积Table 3Common peaks relative peak area of the fingerprints of 14 batches of Poriae Cutis and White Poria decoction pieces样品Sample1S10.3671.6720.6835.3000.8290.1500.4020.6440.4371.0000.3415.8092.2641.0050.677S20.4271.4951.1617.0301.062 0.2120.375 0.5210.6861.0000.5155.0012.2711.823S30.5611.5191.0875.3011.5110.2300.4770.6160.5661.000 0.343 6.2432.4161.438S40.5361.9051.4278.6991.6920.2640.4000.620 0.6921.0000.4586.0192.4891.750S50.4712.5480.9789.4471.2770.2400.4000.5270.523 1.0000.4467.7492.8931.6250.928S60.4841.7191.2336.9721.304 0.2230.3790.4770.6401.0000.4755.5142.2771.406Relative peak area of common peaks234567891011121314151.0440.9461.0570.8461474续表 3(Continued Tab.3)样品Sample1S70.4882.1251.2079.5231.2440.2070.3720.530 0.5801.000 0.4576.3042.6462.007S80.487 1.7861.1057.3021.1970.2430.3480.5230.5571.0000.3716.329 2.5861.668S90.420 2.4480.8699.2031.2890.2090.3940.570 0.4761.0000.3265.7252.2661.237平均值0.4711.913 1.0837.6421.2670.220 0.3940.5590.5731.0000.4156.0772.4561.5510.941MeanS100.0790.3280.3310.3100.4900.1990.0770.4220.4931.0000.3020.2880.1370.022S110.0830.2560.4510.1960.5210.2270.0900.319 0.5801.0000.403 0.4210.2040.0170.036S120.1150.3150.890 0.7860.6920.3250.0490.258 0.5861.0000.1260.8070.350 0.190S130.0600.1800.513 0.163 0.512 0.2660.0650.2510.6101.0000.3030.3110.1540.023S140.089 0.2300.5640.2600.538 0.2170.0390.291 0.609 1.0000.210 0.2600.1370.025平均值0.0850.2620.5500.343 0.550 0.247 0.064 0.308 0.5751.0000.2690.4180.1960.055Mean天然产物研究与开发共有峰相对峰面积Relative peak area of common peaks23Vol.354567891011121314151.1641.0310.7790.0310.1410.0380.0380.0572.4.3相似度分析采用“中药色谱指纹图谱相似度评价系统（2 0 12 版）”软件计算14批茯苓样品HPLC指纹图谱的相似度（见表4）,由结果可知,茯苓皮样品相似Table 4Similarities of 14 batches of Poriae Cutis and White Poria decoction pieces样品S1SampleS11.0000.975 0.994 0.9740.9880.9860.9700.9920.9640.508 0.5060.7330.439S20.9751.0000.9720.9980.9910.997S30.994S40.9740.9980.9701.0000.9930.998S50.9880.9910.9790.9931.0000.9960.9940.9970.9920.4670.451S60.986S70.970 0.9970.9620.9980.9940.9951.000S80.9920.9950.9880.9940.9970.9980.9921.0000.9850.4930.484S90.964S100.508S110.5060.491S120.733S130.439S140.4570.4670.4880.462度在0.951 0.998，白茯苓样品相似度在0.8 940.993,表明不同产地同一药用部位茯苓样品相似度较好;茯苓皮与白茯苓之间相似度在0.38 9 0.7 6 5,说明不同药用部位的茯苓指纹图谱有明显区别。表414批茯苓皮与白茯苓饮片的相似度S2S30.9721.0000.9970.9820.9980.9890.9510.5070.5270.5020.4670.5340.4820.4510.744 0.7650.7410.7050.4380.4720.429S40.9700.9790.9960.9940.9920.9900.9960.9850.5150.4710.5030.4480.7530.7100.7350.3900.4470.4170.476S5S60.9821.0000.9950.9980.9900.5150.5030.9920.9960.4711.0000.4770.4930.4770.4840.4450.7010.8940.9070.3930.4250.4270.4500.4260.9760.9820.9200.993S70.9970.9620.9880.9510.9980.9940.9940.5020.4820.7050.3900.7530.4470.4480.7100.3930.7350.4250.4450.7011.0000.9830.8940.9720.9831.0000.9070.9911.0000.9020.3890.9720.9910.9021.000S80.9950.9890.5070.4910.5270.534S9S10S11S120.7440.4380.7650.4720.7410.4290.389S13S140.4570.4670.4880.4620.4170.4760.4270.4500.4260.9760.9820.9200.9931.0002.5各产地茯苓皮与白茯苓样品的聚类分析聚类分析(CA)是一种按“物以类聚”原则通过将观察对象依据某些特征分类的多元统计分析手段,从而建立样本与样本之间的相似关系或亲疏关系，目前在中药方面的应用较广泛,如真伪鉴别、品种分类及质量评价等16 。采用SPSS26.0软件，以14批不同产地不同药用部位茯苓样品的15个共有峰峰面积为变量,用组间连接法、欧式距离为度量标准进行聚类,聚类结果见图4。由图4可知,14份茯苓样品被分成了2 大类：第一大类为茯苓皮（S1S14Vol.35S9）;第二大类为白茯苓（S10S14）,这与样品的性状鉴别结果吻合,表明不同药用部位间茯苓样品的差异较大,观察“2.4.2”项下各共有峰的相对峰面积均值可发现,茯苓皮中除峰6 和峰9外其他共有10S3S6S5S9S1S8S2S4S7S10S11S13S14S12图414批不同产地茯苓皮和白茯苓样品聚类分析树状图Fig.4 Cluster analysis of 14 batches of Poriae Cutis and White Poria samples from different regions2.6各产地茯苓皮与白茯苓样品的主成分分析将不同产地茯苓皮与白茯苓样品各共有峰峰号对峰面积作折线图（见图5），从图可知在本实验条80006000400020000123456 78910111213 1415共有峰Commonpeaks图514批不同产地茯苓皮和白茯苓样品峰面积折线图Fig.5 Peak area line chart of 14 batches of Poriae Cutis and White Poria samples from different regions2.6.1主成分结果以标准化后的各共有峰峰面积为变量进行PCA分析。表5为主成分结果,前2 个主成分解释了全部方差的90.38 7%，能够反映茯苓皮与白茯苓指纹图谱中共有峰的大部分信息，结合碎石图（见图6)可知,前2 个主成分特征值相对较大，而其他主成分之间比较平缓,因此提取前两个主成分并标记为Y和Y2。图7 为14批茯苓样品的PCA散点得分图,结果表明,14批样品可分为2 类,I类样品为S1-S9，II 样品为 S10-S14,该现象与 CA分析结果基本一致。占慧慧等：茯苓皮与白茯苓中三类成分的HPLC指纹图谱与化学模式识别研究01475色谱峰均高于白茯苓,提示各成分含量的高低可能是造成不同药用部位药材差异的主要原因。此外白茯苓样品按产地分为两类：I类样品为S10和S11，产自湖南;II类样品为S13和S14,产自云南。515下茯苓样品峰面积数据拟合度较小，样本组间差异较大,故采用无监督的主成分分析(PCA)方法,使用的分析软件为SPSS 26.0和SIMCA14.1。一51S2S3S455-S6S7S8-S9*-S10511-S12S13TT20TT2.6.22主成分表达式根据标准化后的各个共有峰变量X1-15占主成分的权重（见表6），得到Yi、Y的线性组合：Y=0.229X,+0.111X,+0.268X,-0.251X4+0.295Xs+0.295X。+0.18 9X,+0.2 8 2 Xg+0.2 99X，+0.300X1o+0.279Xi,-0.257Xi2-0.233X13-0.265X14-0.251Xis，Y,=0.331X,+0.50 4X,-0.0 91X,-0.304X4+0.094X，+0.0 15X。+0.36 0 X,+0.191X。+0.0 41X+0.0 7 7 Xi o +0.157 Xu +0.290X12+0.360X13+0.231Xi4+0.265Xis o25T1476成分Component1234天然产物研究与开发表5茯苓饮片样品的主成分结果Table 5Principal component results of decoction pieces samples made from P.cocos sclerotium初始特征值Initial eigenvalues方差百分比合计VarianceTotalpercentage10.82372.1532.73518.2340.7725.1490.3502.330Vol.35提取载荷平方和Extract the sum of squared loads累积百分比合计CumulativeTotalpercentage72.15390.38795.53697.866方差百分比Variancepercentage10.82372.1532.73518.234累积百分比Cumulativepercentage72.15390.3871086420123 4567 8910 11121314 15共有峰Common peaks图6 茯苓饮片样品PCA碎石图Fig.6 PCA scree plot of decoction pieces samples