Pharmacokinetics of aconitine as the targeted marker of Fuzi (Aconitum carmichaeli) following single and multiple oral administrations of Fuzi extracts in rat by UPLC/MS/MS

Ethnopharmacological relevance

Fuzi, which is the processed lateral roots of Aconitum Carmichaeli. Debx and is widely distributed over the southwest provinces of China, is recognised for its anti-inflammatory and analgesic effects.

Aim of the study

The pharmacokinetic properties of Fuzi are inadequately understood. Aconitine, the primary highly toxic ingredient of Fuzi, is well known as the target marker of Fuzi. The purpose of the present study is to investigate the pharmacokinetic behaviours of aconitine in vivo following single and multiple administrations of processed Fuzi extracts and to compare the pharmacokinetic characteristics of aconitine after administrations of pure aconitine or Fuzi extracts as well as compare the difference at single dose and multiple doses. The in vitro aconitine protein binding in plasma through equilibrium dialysis was also examined.

Methods

A high performance liquid chromatography (HPLC) method was developed for the determination of aconitine in Fuzi crude extracts and a fast ultra performance liquid chromatography-tandem mass spectrometry (UPLC/MS/MS) was developed to investigate the pharmacokinetic behaviour of aconitine as the targeted marker of Fuzi.

Results

The absolute bioavailability (F %) after the administration of 0.5 mg/kg aconitine and Fuzi extract (0.118 mg/kg aconitine) in rat was 8.24 ± 2.52% and 4.72 ± 2.66%, respectively. Aconitine absorption was very fast at the t_max 30.08 ± 9.73 min for pure aconitine and 58.00 ± 21.68 min for Fuzi extract administration. Aconitine was also eliminated rapidly with a short half-life (i.v., 80.98 ± 6.40 min) and a low rate of protein bounding (23.9–31.9%). No significance was observed on all the pharmacokinetics parameters following the single and multiple doses of pure aconitine (ANOVA, p > 0.05). However, the absorption of aconitine after multiple administrations of Fuzi extract was much faster than that of a single dose (t_max: 58.00 ± 21.68 vs. 20.00 ± 8.66 min, p < 0.05), and the area under the plasma concentration–time curve (AUC) was higher than that of a single dose.

Conclusions

The pharmacokinetic behaviour of processed Fuzi was determined in this paper. The aconitine has low bioavailability. No variation in the pharmacokinetic behaviours of pure aconitine was observed after single and multiple administrations. In contrast, multiple administrations of processed Fuzi extract could result in variations in its pharmacokinetic behaviour in AUC and t_max indicating that multiple dose might increase the bioavailability of aconitine, which may result in its toxicity. In addition, aconitine has a low protein bounding (23.9–31.9%), resulting in its rapid elimination.     

灯心草煅炭前后UPLC指纹图谱研究及菲类成分含量测定

目的 建立灯心草及其炮制品灯心炭超高效液相色谱法（UPLC）指纹图谱及灯心草中3个菲类成分含量测定方法,比较灯心草炮制前后质量差异。方法 采用UPLC建立灯心草、灯心炭指纹图谱,并进行相似度评价,采用方差分析、聚类分析（CA）、主成分分析（PCA）对结果进行评价。结果 建立了灯心草与灯心炭UPLC指纹图谱,生品标定15个共有峰,炭品标定16个共有峰,与相对应的对照指纹图谱相似度均大于0.89;通过CA将灯心草和灯心炭聚为两类,PCA筛选出4个主因子;确定了灯心草3个菲类成分的UPLC定量方法。结论 建立的指纹图谱方法稳定可靠,炮制后灯心草化学成分发生变化,为灯心草临床用药及质量控制提供参考。     

基于UPLC特征图谱结合多模式识别和色度值的槐角炮制前后差异研究

目的 测定槐角炮制前后的超高效液相色谱（UPLC）特征图谱和色度值,结合不同模式识别方法比较槐角炮制前后的差异性,为槐角和蜜槐角的质量控制及评价提供参考。方法 采用UPLC法对槐角炮制前后样品进行测定,按《中药色谱指纹图谱相似度评价系统（2012版）》对色谱图进行匹配生成槐角炮制前后的UPLC特征图谱,并对炮制前后的色度值（L、a、b）进行测定,通过相似度分析、对照品比对、方差分析、聚类分析、主成分分析（principal component analysis,PCA）和正交偏最小二乘判别分析（orthogonal partial least squares discriminant analysis,OPLS-DA）对槐角炮制前后进行多模式识别研究。结果 建立了槐角炮制前后的UPLC特征图谱,槐角和蜜槐角分别确定了23、24个共有峰,指认2、15、16、20、21号峰为没食子酸、芦丁、染料木苷、槲皮苷、槐角苷,炮制后新增成分24号峰为5-羟甲基糠醛;相似度分析结果显示槐角炮制前后相似度均大于0.98,聚类分析结果显示槐角炮制前后区分不明显,PCA可以将槐角和蜜槐角大致分为2类,OPLS-DA分析可将槐角和蜜槐角明显区分为2类,且结果显示1、2、10、22、23、24号峰是引起槐角和蜜槐角间成分差异的主要标志性成分,与方差分析结果一致;色度值ΔE范围为5~13,炮制前后色差可被肉眼识别。结论 建立的多指标特征图谱和色度值鉴别方法对槐角药材及其炮制品的质量控制及整体性评价具有重要意义。     

Chondroprotective activity of a detoxicated traditional Chinese medicine (Fuzi) of Aconitum carmichaeli Debx against severe-stage osteoarthritis model induced by mono-iodoacetate

Ethnopharmacological relevance: Fuzi is an effective but toxic traditional Chinese medicine (TCM) derived from Aconitum carmichaeli. In our previous study, detoxicated Fuzi (d-Fuzi) has been originally developed with no toxicity but significant efficacy. However, whether d-Fuzi can be used for therapy of osteoarthritis (OA), remain unknown.Materials and methods: Severe OA model was established by intra-articular mono-iodoacetate (MIA) injection (1.25 mg) into rats and orally treated with 2 g/ml d-Fuzi at a dosage of 7 ml/kg body weight for 28 days. In vivo, the articular radiographic and histopathologic analyses were performed to qualitatively assess the chondroprotective effect of d-Fuzi, followed by quantitative measurements of bone density and Mankin scores. In vitro, such effect on chondrocyte viability after MIA attack was evaluated. Hybrid quadrupole time-of-flight mass spectrometry (QTOF-MS) was performed for chemical analysis of d-Fuzi.Results: d-Fuzi was demonstrated to possess chondroprotective activity on MIA-induced OA model by in vivo preventing the articular degeneration and the reducing of bone density and Mankin score, as well as by in vitro promoting the chondrocyte proliferation and inhibiting the MIA-induced chondrocyte damage. A total of 23 compounds were identified in d-Fuzi, most of which were deduced as the non-toxic derivatives of aconite alkaloids.Conclusions: This is the first report regarding chondroprotective effect and chemical profile of d-Fuzi, originally revealing its great anti-OA potential and thereby providing a promising TCM candidate for OA therapy.     

Development and assessment of a complete-detoxication strategy for Fuzi (lateral root of Aconitum carmichaeli) and its application in rheumatoid arthritis therapy

Ethnopharmacological relevance

Fuzi (lateral root of Aconitum carmichaeli) is a popular traditional Chinese medicine well known for its both therapeutic and high-toxic activities. Its toxic alkaloid ingredients, mainly aconitine, mesaconitine, and hypaconitine, are responsible for the high toxicity. However, to date, no detoxication strategy is available to completely eliminate Fuzi's toxicity, and, whether Fuzi's efficacy could be kept after detoxication, remain unknown and debatable.

Materials and methods

The purpose of this study was to establish and validate a complete-detoxication strategy for Fuzi via acute toxicity test, to clarify the detoxication mechanism by HPLC and titrimetric analyses, and to evaluate the therapeutic effect of detoxicated Fuzi on adjuvant arthritis (AA). Three processed Fuzi (Bai-fu-pian) with 30-min, 60-min, and 120-min decoctions, respectively, named dBfp-30, dBfp-60, and dBfp-120, were prepared for this study. For the acute toxicity test, their oral doses to male and female Kunming mice were up to 70–190 g/kg body weight, and their toxicological profiles were evaluated by median lethal dose (LD₅₀), maximal tolerance dose (MTD), minimal lethal dose (MLD), no-observed-adverse-effect-level (NOAEL), and time–concentration–mortality (TCM) modeling methods using a 14-day schedule with up to five doses. The HPLC analysis was performed to determine the detoxication-induced changes in composition and amount of aconitine, mesaconitine and hypaconitine in Fuzi, whilst the titrimetric method was adopted to estimate the amount changes of Fuzi's total alkaloids. AA model was established by incomplete Freund's adjuvant injection in Wistar rats, and the animal's physiological (body weight, food intake, etc.), clinical (hind paw volume), and immunological (IL-1 and TNF-α) parameters were assessed as markers of inflammation and arthritis.

Results

With increasing decoction time, the acute toxicity of detoxicated Fuzi became decreased in the following order: dBfp-30 (LD₅₀ of 145.1 g/kg; MTD of 70 g/kg; MLD of 100 g/kg; NOAEL of 70 g/kg) >dBfp-60 (too large LD₅₀; MTD of 160 g/kg; MLD of 190 g/kg; NOAEL of 100 g/kg) >dBfp-120 (no LD₅₀; unlimited MTD; unlimited MLD; NOAEL of 130 g/kg). dBfp-30 and dBfp-60 displayed the toxicity at a dose-dependent manner with maximum mortalities reaching 100% and 50% respectively, whereas no mortality or signs of intoxication was induced by dBfp-120. The chemical analyses revealed a dramatic reduction of the toxic alkaloids as well as total alkaloids in Fuzi after the detoxication, from which no level of aconitine and only minimum residual of mesaconitine (0.56±0.02 μg/g) and hypaconitine (8.73±0.13 μg/g) were detected in dBfp-120. However, no significant difference of total alkaloid amount was found among dBfp-30, dBfp-60, and dBfp-120 (P>0.05), suggesting an equivalent conversion from toxic alkaloids to its non-toxic derivants in dBfp-120. Further, also no significant differences were seen among dBfp-30, dBfp-60, and dBfp-120 for the therapeutic effects on physiological, clinical, and immunological parameters in AA rat, indicating that dBfp-120 is of non-toxicity and efficacy.

Conclusions

A complete-detoxication strategy has been developed successfully for ensuring the safe and effective use of Fuzi. The detoxication mechanism associated with elimination of toxic alkaloids has kept Fuzi's efficacy, indicating a non-interdependent relationship between its efficacy and toxicity. This is the first report on such an optimal detoxication strategy and on the application of detoxicated Fuzi in AA. It may provide in depth understanding to the toxicological and pharmacological profiles of Fuzi and further benefit the herbal drug development with safety and efficacy for disease especially RA therapy.     

炉甘石生品及炮制品X射线衍射指纹图谱分析

目的建立炉甘石生品及炮制品的X射线衍射(XRD)指纹图谱;比较不同炮制方法对药材主要成分的影响;测定炮制品中ZnO含量。方法采用XRD法对10批炉甘石生品及炮制品进行分析,分别建立了生品及炮制品的指纹图谱;对6种不同炮制方法进行比较;运用K值法测定了炮制品中ZnO的含量。结果分别建立了炉甘石生品和炮制品的XRD指纹图谱,其中生品XRD指纹图谱有23个共有峰,炮制品XRD指纹图谱有10个共有峰,经煅烧炮制之后生品的ZnCO_3特征峰转变为ZnO的特征峰;煅烧炮制品中ZnO的含量均超过56%。结论 XRD指纹图谱可用于炉甘石生品及炮制品的鉴定与分析,为炉甘石生品及炮制品的质量评价提供了新的可靠方法。     

基于UPLC指纹图谱结合多模式识别的荆芥穗炮制前后差异研究

目的 测定荆芥穗炮制前后的超高效液相色谱法（UPLC）指纹图谱,结合不同模式识别方法比较炮制前后差异,为荆芥穗和荆芥穗炭质量控制及评价提供参考。方法 采用UPLC对荆芥穗炮制前后样品进行测定,生成对照指纹图谱,通过相似度分析、对照品指认、方差分析、聚类分析（HCA）、主成分分析（PCA）和正交偏最小二乘法-判别分析（OPLS-DA）对荆芥穗炮制前后进行多模式识别研究。结果 分别建立了荆芥穗和荆芥穗炭的UPLC指纹图谱,确定了13个共有峰,指认4、9、11、13号峰分别为木犀草苷、迷迭香酸、胡薄荷酮、木犀草素;相似度结果显示,炮制前后相似度均大于0.9,HCA、PCA、OPLS-DA结果可明显将荆芥穗和荆芥穗炭区分为2类,2、4、7~12号峰是引起荆芥穗和荆芥穗炭间成分差异的主要标志性成分,与方差分析结果一致。结论 建立的多指标指纹图谱鉴别方法对荆芥穗药材及其炮制品的质量控制及整体性评价具有重要意义。     

Toxicity of Five Herbs in Aconitum L. on Tetrahymena thermophila Based on Spectrum-effect Relationship

Objective To explore the active components with toxic effects in five Aconitum L.herbal medicines on Tetrahymena thermophila.Methods The fingerprints of five Aconitum L.herbal medicines were established by ultra-high performance liquid chromatography(UPLC)and the toxicity was evaluated by using a TAM Air Isothermal Calorimeter on Tetrahymena thermophila SB110.Results By analyzing the spectrumeffect relationships between UPLC fingerprints and toxic effects,the active components which had the toxic effects were obtained.Conclusion This work provides a general model of the combination of UPLC and microcalorimetry to study the spectrum-effect relationships of the five Aconitum L.herbal medicines,which could be used to evaluate the toxic effects and analyze the principal toxic components of the five Aconitum L.herbal medicines.On the whole,this result provides the experimental basis for the safe use of the five Aconitum L.herbal medicines in clinic.     

苍术炒焦前后GC指纹图谱的比较

目的:建立生苍术和焦苍术挥发油的GC指纹图谱,阐明苍术炒焦前后挥发油部位化学成分的变化规律。方法:采用水蒸气蒸馏法制备挥发油;建立气相色谱指纹图谱,检测条件为HP-5石英毛细管柱(0.32 mm×30 m,0.25μm),载气(N2)流速0.8 m L·min-1,进样口温度240℃,进样量1μL,分流进样,分流比20∶1,检测器温度260℃,柱温采用程序升温模式;并对炒焦前后的GC指纹图谱进行比较与分析。结果:在苍术炒焦前后挥发油的GC指纹图谱中有25个共有峰,各共有峰相对保留时间基本相同,但相对峰面积发生了显著变化;其中15~17,19,23~25号峰的峰面积降低率均20%;16号与17号色谱峰的峰面积比值在生品中均3.0,而在焦品中却均3.0;19号与23号色谱峰的峰面积比值在生品中均3.0,而在焦品中却均3.0。鉴定16号峰为β-桉叶醇,19号峰为邻苯二甲酸二异丁酯,23号峰为白术内酯Ⅱ,24号峰为白术内酯Ⅰ,25号峰为白术内酯Ⅲ。结论:苍术生品和焦品的GC色谱特征峰差异明显、结果稳定、专属性强,可作为二者的定性鉴别特征之一,这为阐释焦苍术的降燥减毒机制提供了实验依据。     

The effects of Rhizoma Zingiberis on pharmacokinetics of six Aconitum alkaloids in herb couple of Radix Aconiti Lateralis−Rhizoma Zingiberis

Ethnopharmacological relevance

Radix Aconiti Lateralis (Fuzi in Chinese, derived from the lateral roots of Aconitum Carmichaeli Debx.) is widely used for the treatment of heart failure, internal cold, arthralgia, diarrhea and edema for thousands of years. It was usually prescribed in combination with Rhizoma Zingiberis (Ganjiang in Chinese, derived from the dry rhizome of Zingiber officinale Rosc.) to decrease toxicity and increase efficacy.

Aim of the study

In order to investigate the influence of Rhizoma Zingiberis on pharmacokinetics of six Aconitum alkaloids, i.e. aconitine (AC), hypaconitine (HA), mesaconitine (MA), benzoylaconine (BAC), benzoylhypaconine (BHA) and benzoylmesaconine (BMA), in Fuzi−Ganjiang herb couple, the comparative pharmacokinetics of six Aconitum alkaloids after oral administration of Fuzi and Fuzi−Ganjiang aqueous extract was carried out.

Materials and methods

A sensitive, specific and rapid LC−MS/MS method was developed to determine the six analytes in plasma. Then the rats were randomly divided into two groups and orally administered with Fuzi and Fuzi−Ganjiang aqueous extract. At designated time points after oral administration, the concentrations of the six Aconitum alkaloids in rat plasma were determined, and main pharmacokinetic parameters were investigated using 3P97 (Practical Pharmacokinetics Program Version 1.0).

Results

Comparing with Fuzi group, both T_1/2 and AUC_0−t of AC and HA decreased (P<0.05), while T_1/2, AUC_0−t and C_max of BAC, BHA increased (P<0.05) in Fuzi−Ganjiang group, which indicated that Ganjiang could promote the elimination of AC and HA and enhance the absorption of BAC, BHA and BMA.

Conclusion

The differences of pharmacokinetics of Aconitum alkaloids in rat plasma could support those of pharmacologics and toxicity in previous reports between Fuzi and Fuzi−Ganjiang herb couple. The results might be helpful in explaining the mechanism of combination of Fuzi−Ganjiang to decrease toxicity and increase efficacy.     

榼藤子生品与炮制品HPLC指纹图谱研究

目的:建立榼藤子生、炮制品HPLC指纹图谱的评价方法,分析各样品指纹图谱的变化。方法:采用HPLC-UV分别对榼藤子生品和炮制品的指纹图谱进行研究,用HPLC-ESI-MS对其生品共有峰进行鉴定。结果:标示出生品16个共有峰,炮制品21个共有峰,应用HPLC-MS和对照品保留时间的对应初步鉴定了其中9个共有峰,主要为榼藤子三萜皂苷类和其他糖苷类成分的吸收峰。结论:榼藤子生品炮制后,60%甲醇提取液中部分化学成分含量有不同程度的下降或上升,指纹图谱的主峰数目也有变化,说明该法可以较全面地反映榼藤子炮制前后化学成分的差异,用于控制生品和炮制品的内在质量。     

苍术麸炒前后的专属性HPLC特征指纹图谱

目的:比较苍术麸炒前后专属性HPLC特征图谱,明确特征性图谱差异性,为苍术生品和麸炒品提供专属性质量评价指标。方法:取10批生苍术和对应麸炒苍术饮片的正己烷提取物进行HPLC分析,流动相色谱乙腈-水梯度洗脱,流速1.0 m L·min~(-1),检测波长254 nm。以苍术素色谱峰为参照峰,构建了10批苍术生品和麸炒品的HPLC指纹图谱的共有模式。结果:苍术生品和麸炒品HPLC中有26个共有峰,各共有峰相对保留时间基本相同,RSD均0.5%;但相对峰面积有明显差异:第47.36 min和第46.35 min色谱峰的峰面积比值在生品中1,但在麸炒品中却1;第51.70 min和第49.25 min色谱峰的峰面积比值在生品中1,但在麸炒品中却1。结论:苍术生品和麸品的色谱特征峰差异明显、结果稳定、专属性强,可作为生苍术和麸炒苍术饮片的定性鉴别特征之一。     

款冬花不同炮制品的HPLC指纹图谱比较

目的:采用HPLC指纹图谱技术,综合运用相似度评价、聚类分析(CA)及主成分分析(PCA)对款冬花炮制前后化学成分变化进行研究。方法:采用HPLC-DAD建立10个产地款冬花不同炮制品的指纹图谱,流动相乙腈-0.03%三氟乙酸水溶液梯度洗脱,检测波长240 nm;运用"中药色谱指纹图谱相似度评价系统"(2004A版)软件,CA及PCA综合探索款冬花不同炮制品的质量及成分变化规律。结果:建立了不同炮制品的HPLC共有模式,款冬花生品、蜜炙品、甘草炙品指纹图谱的相似度分别为0.867~0.991,0.785~0.979,0.785~0.980。CA将不同炮制品均分为4类,与相似度评价结果基本一致,表明不同产地间款冬花炮制品具有一定的相似性和稳定性。PCA筛选出方差累计值达74.230%的2个主成分,并得到与之相关的成分群,不同产地炮制品的综合得分情况表明该成分群在生品、蜜炙品及甘草炙品中稳定存在。结论:指纹图谱技术结合相似度评价,CA,PCA能对款冬花炮制品的质量进行系统评价,可为其他中药炮制品的评价提供参考。     

近红外光谱结合主成分分析和聚类分析鉴别炉甘石生品、伪品和炮制品

目的:利用主成分判别分析和聚类分析建立炉甘石生品、伪品和炮制品的近红外光谱鉴别模型。方法:采集炉甘石生品、伪品和炮制品的近红外光谱,每类样品随机划分为训练集和测试集。对光谱预处理方法和建模谱段进行筛选,分别建立主成分判别分析模型及聚类分析模型。结果:光谱经一阶导数预处理,主成分判别分析模型的特征谱段为4 800~4 000 cm~(-1),聚类模型的特征谱段为7 300~7 000,4 800~4 000 cm~(-1)。在主成分判别分析模型中,预测准确率94.34%;在聚类模型中,模型的预测准确率96.23%。结论:所建立的近红外主成分判别分析模型和聚类分析模型均可用于炉甘石生品、伪品和炮制品的鉴别。     

不同产地穿心莲的含量测定、化学指纹图谱及抑菌活性评价

目的：从化学和生物两方面综合考察不同产地穿心莲药材品质差异。方法：采用微量量热法比较13批不同产地穿心莲药材的抗痢疾杆菌生物活性。同时建立穿心莲的UPLC指纹图谱,采用偏最小二乘法（PLS）回归分析抑菌活性与化学成分之间的相关性。结果：13批不同产地的穿心莲药材使痢疾杆菌生长代谢的时间延长,生长速率常数减小,表明不同产地穿心莲对痢疾杆菌的生长代谢均有不同程度的抑制作用,且以西双版纳产地的穿心莲药材抑菌效果最好。此外,通过偏最小二乘法回归分析找到了四个与抑菌活性密切相关的化学成分。结论：不同产地穿心莲药材质量差异较大,云南、海南、广西和四川综合质量较好。生物活性测定结合化学指纹图谱评价药材质量的方法,可准确、可靠地评价不同产地穿心莲药材的质量,为其质量控制提供依据。     

栀子不同炮制品标准汤剂的对比研究

目的 研究栀子不同炮制品标准汤剂质量指标的差异。方法 测定19批栀子、炒栀子和焦栀子标准汤剂出膏率、指标成分栀子苷的含量和转移率,并建立标准汤剂指纹图谱,计算指纹图谱相似度,采用对照品比对的方式对共有峰进行确证,以指纹图谱共有峰的峰面积为变量,进行正交偏最小二乘法-判别分析（OPLS-DA）,寻找差异性标志物。结果 标准汤剂的出膏率排序焦栀子>栀子>炒栀子;栀子苷的含量排序栀子>炒栀子>焦栀子;栀子苷的转移率排序焦栀子>栀子>炒栀子。栀子、炒栀子和焦栀子指纹图谱均标识出7个共有峰,指认出其中5个成分,栀子不同炮制品标准汤剂指纹图谱的相似度均大于0.95;OPLS-DA将栀子、炒栀子和焦栀子分为两类,并找到5个差异性成分,分别为峰6、峰5、峰7、峰3和峰2,表明随着炒制程度的加深,上述化学成分在标准汤剂中的含量逐渐降低。结论 可为栀子、炒栀子和焦栀子配方颗粒及其经典名方复方制剂的研究与开发提供参考。     

基于指纹图谱探讨吴茱萸不同炮制品成分的差异

目的:探讨吴茱萸不同炮制品化学成分的差异.方法:甘草和开水作为辅料分别炮制吴茱萸,获得2个吴茱萸炮制品,并建立吴茱萸药材和吴茱萸炮制品3种样品各10批的指纹图谱,采用中药色谱指纹图谱相似度评价系统提取相关信息,用SPSS进行数据分析.结果:以10批吴茱萸生品所生成的对照图谱为参照,吴茱萸生品、甘草水泡吴茱萸及开水泡吴茱萸样品的相似度均＞0.9.对30个吴茱萸样品全匹配色谱峰的聚类分析获得22个特征峰,并对此22个特征峰的峰面积进行分析,结果表明有8个特种峰经过炮制后峰面积显著性下降,另有6个特征峰经甘草水泡制后峰面积显著降低,而吴茱萸内酯及其吴茱萸碱的峰经甘草水泡制后峰面积显著性增加.结论:吴茱萸不同炮制品及吴茱萸生品的特征峰所代表的成分有显著性的差异.     

微量热法对不同生长年份黄连品质的评价

目的尝试构建基于生物热力学表达的中药品质评价方法体系。方法以不同生长年份黄连为例,利用微量热法,测定黄连作用于大肠杆菌生长代谢过程中的热功率图,建立生物热动力学模型,分析热动力学参数、生物活性与化学成分之间的相关性。结果在不同生长年份黄连的作用下,大肠杆菌生长代谢的热功率图不尽一致,呈现较明显的指纹特征;与正常对照组比较,不同生长年份黄连均能不同程度地使大肠杆菌生长速率常数减小,传代时间延长,最大产热功率降低,说明不同生长年份黄连对大肠杆菌生长代谢均有不同程度的抑制作用,其中以含总生物碱最高的四年生黄连的抑制作用最强。结论不同生长年份黄连作用于大肠杆菌生长代谢过程的部分热动力学参数与热生物活性和化学成分之间具有显著的相关性;是具有实时、在线、微量、高效、高通量、普适性好等特点的生物热力学方法和指标,既可以间接地反映黄连的生物活性,也可作为评价中药品质的一种新方法。     

红花UPLC法特征指纹图谱研究

目的建立红花Carthamus tinctorius超高效液相色谱(UPLC)指纹图谱,为评价不同产地和不同品种红花的质量提供参考。方法对产地来自新疆和河南共34批红花样品应用UPLC建立其指纹图谱,按"中药指纹图谱相似度评价系统"建立对照图谱,并进行相似度评价。运用SAS 9.30和Matlab软件对新疆不同品种和不同产地红花的UPLC指纹图谱特征峰进行聚类分析和主成分分析。结果建立了红花药材的专属性UPLC指纹图谱,确立了26个共有峰;30批新疆红花样品和4批河南红花样品质量差异较大,新疆不同产地间和同一产地不同品种间具有一定差异。聚类分析和主成分分析均实现了新疆红花指纹图谱差异的识别。结论该方法简便、高效、可靠,为红花药材的质量控制提供了有效的快速评价方法。     

多波长覆盖融合指纹图谱评价不同产地川芎药材差异性

目的:建立10批不同产地川芎药材多波长覆盖融合指纹图谱,并对共有峰进行初步指认,通过指纹图谱比较不同产地川芎药材的差异性,为其质量控制和药材鉴别提供依据。方法:采用超高液相色谱法建立不同产地川芎药材的指纹图谱;采用全时段多波长融合技术对dif数据进行多波长融合;采用Spss 19.0数据统计软件,对不同产地川芎药材的共有峰进行聚类分析,区别其差异;采用Q-TOF-MS法对指纹图谱中的共有峰进行指认。结果:建立了川芎药材的多波长融合指纹图谱,确定了20个共有峰并对其中的8个峰进行了指认,通过聚类分析和相似度评价可知云南川芎与其他产地的川芎相比存在着一定的差异性。结论:该指纹图谱重复性良好,能够区分不同产地川芎药材,可作为川芎药材质量评价的一种手段。