HPLC Fingerprint and Chemical Pattern Recognition of Wild and Cultivated Millettia speciosa Champ.

[Objectives]To establish HPLC fingerprints of wild and cultivated Millettia speciosa Champ.,and identify medicinal materials combined with chemical pattern recognition methods,and provide a reference system for the identification and quality control of M.speciosa from different sources.[Methods]20 batches of M.speciosa from different sources were determined by HPLC method,and the similarity analysis and evaluation were performed using the Similarity Evaluation System of Traditional Chinese Medicine Chromatographic Fingerprints(2012 edition).Principal component analysis(PCA)and least partial squares method-discrimination analysis(PLS-DA)were used to conduct chemical pattern recognition research on wild and cultivated M.speciosa.[Results]The HPLC fingerprints of wild and cultivated M.speciosa were established,10 common peaks were calibrated,and the similarity of 20 batches of samples was greater than 0.9;PCA can better classify M.speciosa from different sources into 2 categories,and PLS-DA can completely distinguish between wild and cultivated M.speciosa.[Conclusions]The established M.speciosa fingerprint,combined with chemical pattern recognition methods,can effectively distinguish between wild and cultivated M.speciosa,so it can provide a reference for quality control and evaluation of M.speciosa.

GC-MS Analysis of Liposoluble Constituents from Buds,Flowers and Fruits of Millettia speciosa Champ.

[Objectives] To study the material foundation of liposoluble constituents from buds,flowers and fruits of Millettia speciosa Champ.and provide a reference for the development of flowers and fruits of Millettia speciosa Champ.[Methods]The liposoluble constituents were extracted from buds,flowers and fruits of Millettia speciosa by Soxhlet extraction and solvent extraction method,and analyzed by GC-MS.[Results]24 compounds were identified from the liposoluble constituents of buds,accounting for 88. 31 % of the total liposoluble constituents,mainly comprising alkanes and olefins compounds( 52. 00%) and alcohols compounds( 17. 46%); 29 compounds were identified from the liposoluble constituents of flowers,accounting for 91. 38 % of the total liposoluble constituents,mainly comprising alkanes and olefins compounds( 60. 64%) and alcohols compounds( 17. 17%); 32 compounds were identified from the liposoluble constituents of fruits,accounting for 80. 01 % of the total liposoluble constituents,mainly comprising alkanes and olefins compounds( 32. 56%),phenyl and its derivatives compounds( 22. 46%) and fatty acids compounds( 12. 54%). 6 compounds were common in buds,flowers and fruits. [Conclusions] Although there were some differences in liposoluble constituents from flowers,fruits,leaves and roots of Millettia speciosa Champ.,the different parts of Millettia speciosa Champ. had development value.

Development and Processing of Millettia speciosa Instant Tea

This paper analyzed the effect of raw material crushing fineness,cooking time,ethanol content during ethanol precipitation and other factors on the preparation of raw material extract from Millettia speciosa Champ. instant tea. The raw materials of Millettia speciosa Champ.,Philippine flemingia root and radix fici simplicissimae were crushed into 10 mesh or finer powder,and cooked for 60 min. During ethanol precipitation,the ethanol content was about 50% to 70%,standing 12 h. The ophiopogon root was cooked in 1∶ 15 boiling water for 45 min,and chrysanthemum was leached for 45 min with 1∶ 20 demineralized water at 80 ℃. After concentration,preparation and spray drying,the finished Millettia speciosa Champ. instant tea was created. The detection of each product index indicated that Millettia speciosa Champ. instant tea had good taste and flavor,and there were no heavy metals,harmful substances and excessive microbes,thereby showing that the raw material of Millettia speciosa Champ. instant tea was selected reasonably,the mixture ratio was rational,and the processing technology was of some security,stability and maturity,which provided a theoretical basis for its development and application.

Content Determination of Flavonoids in Radix Millettiae Speciosae from Different Areas of Guangxi

[Objectives] This study aimed to investigate the quality of Radix Millettiae Speciosae at different planting sites in Guangxi. [Methods] The content of flavonoids in Radix Millettiae Speciosae was determined by HPLC. The chromatographic conditions were as follows: column, ZORBAX SB-C_(18); mobile phase, acetonitrile-0.1% glacial acetic acid(42∶58); detection wavelength, 250(formononetin) and 310(maackiain) nm; flow rate, 1.0 mL/min; and column temperature, 30℃. [Results] Under the chromatographic conditions above, formononetin and maackiain could be completely separated from impurities. The standard curve had a good linear relationship(R^2>0.999 9). The precision and stability met analysis requirements. The average recovery rates were 97.53% and 98.54%, respectively, and the RSD values were 2.03% and 1.88%, respectively, indicating that the established method has good reproducibility, durability and accuracy. The content of formononetin was highest in the Radix Millettiae Speciosae from Yongning District, Nanning City, and the content of maackiain was highest in the Radix Millettiae Speciosae from Xichang Town, Hepu County. [Conclusions] The HPLC method established in this study is simple, accurate and stable. It can be used as a quality control method for Radix Millettiae Speciosae. The contents of flavonoids in Radix Millettiae Speciosae from Yongning District, Nanning City and Xichang Town, Hepu County were higher than those from other planting bases.

A Preliminary Study on Anti-inflammatory and Analgesic Effects of Millettia speciosa and Tinpspora sinensis and Their Compatibility

[Objectives]To study the anti-inflammatory and analgesic effects of Millettia speciosa and Tinpspora sinensis and their compatibility.[Methods](i)In the glacial acetic acid writhing experiment,60 SPF-grade Kunming mice were adopted,and the mice were randomly divided into 6 groups at male-female ratio of 1∶1,namely,the blank control group,M.speciosa group,T.sinensis group,M.speciosa compatible with T.sinensis group at the ratio of 1∶2(expressed as 1∶2 compatibility group),M.speciosa compatible with T.sinensis group at the ratio of 1∶1(expressed as 1∶1 compatibility group),and M.speciosa compatible with T.sinensis group at the ratio of 2∶1(expressed as 2∶1 compatibility group),12 mice for each group.Mice of the experimental groups were administered at a dose of 20 mL/kg,and the corresponding concentration of the Chinese medicine extract was given at 1 g/mL.The control group was administered with an equal volume of 0.9%physiological saline,and was intragastrically administered once every 24 h for 14 d.After intragastric administration for one hour on day 14,intraperitoneal injection of 0.5%glacial acetic acid solution was performed to induce pain.(ii)In the hot plate experiment,60 Kunming female SPF mice were adopted,grouped,intragastrically administered with the same glacial acetic acid writhing experiment for 14 d.After intragastric administration for one hour on day 14,the mice were placed on a hot plate apparatus at(55±0.5)℃.to measure the time of licking their hind feet.(iii)In the anti-inflammatory experiment,60 Kunming SPF mice were adopted,grouped,intragastrically administered with the same glacial acetic acid writhing experiment for 14 d.After intragastric administration for one hour on day 14,xylylene was administered to the left ears of mice at a dose of 50μL/piece to induce inflammation.[Results](i)In the glacial acetic acid writhing experiment,compared with the blank control group,the experimental group showed analgesic effects.Specifically,M.speciosa group,T.sinensis group,1∶2 compatibility group,1∶1 compatibility group,2∶1 compatibility group showed significant effect(P<0.05),the writhing inhibition rate was 17.65%,20.59%,29.41%,26.47%,and 44.12%,respectively,and 2∶1 compatibility group showed the most significant analgesic effects.(ii)In the hot plate experiment,compared with the control group,all experimental groups showed analgesic effect.Specifically,M.speciosa group,T.sinensis group,1∶2 compatibility group,1∶1 compatibility group,2∶1 compatibility group showed significant effect(P<0.05),the pain threshold improvement rates were 16.13%,14.55%,14.96%,29.95%,and 58.68%,respectively,and 2∶1 compatibility group showed the most significant analgesic effect.(iii)In the anti-inflammatory experiment,the swelling degree of the 1∶2 compatibility group was significantly different from that of the blank control group,M.speciosa group,T.sinensis group(P<0.05).and 1∶2 compatibility group showed the most significant anti-inflammatory effect.[Conclusions]M.speciosa,T.sinensis,and their compatibility had anti-inflammatory and analgesic effects.The 2∶1 compatibility group had the best analgesic effects,and 1∶2 compatibility group had the best anti-inflammatory effects.

Analysis on Development Potential of Millettia Specisoa Champ Industry in Guangdong Province

In recent years,Millettia specisoa Champ has attracted extensive attention because of its high medicinal and edible value. This paper will summarize the composition and application of Millettia specisoa Champ,analyze the advantages and existing problems in developing Millettia specisoa Champ industry in Guangdong Province according to the demand for Millettia specisoa Champ and the present situation in Guangdong Province,and put forward reasonable countermeasures and suggestions.

牛大力热风干燥工艺参数优化

热风干燥有利于提高鲜制牛大力干燥时的干燥速率和产品品质。为此,对新鲜的牛大力进行单因素热风干燥实验,结果表明:干燥过程中热风温度越高、切片厚度越薄时,牛大力的干燥速率越快;而热风温度过高时,牛大力切片表面会发生褐化和裂纹。结合响应面优化分析方法,建立各因素与指标值之间的回归分析模型。由响应面优化分析和实验结果可知:当热风温度为50~60℃、切片厚度为3~7 mm和热风风速为0.5~1.5 m/s时,牛大力热风干燥的最佳干燥参数为:热风温度60℃,切片厚度5 mm,热风风速1.116 m/s。

2种A型淀粉的消化特性研究

以牛大力淀粉(MSS)和海南红山兰米淀粉(RRS)为研究对象,采用体外模拟消化模型建立一阶消化动力学方程,并对消化后淀粉的微观结构进行表征。结果表明:MSS抗消化能力优于RRS。经体外消化后,牛大力消化淀粉(MSDS)中的直链淀粉含量减少4.95%,而海南红山兰米消化淀粉(RRDS)中的直链淀粉含量增加5.11%。在体外消化过程中,与原淀粉相比,MSDS和RRDS的短程有序结构比例逐渐增大,相对结晶度提高。

UPLC测定不同牛大力样品中刺桐碱的含量

目的:优化牛大力中刺桐碱的提取方法,并建立测定牛大力中刺桐碱含量的超高效液相色谱(UPLC)方法,从而比较不同牛大力中刺桐碱的含量。方法:采用单因素和正交实验优化牛大力中刺桐碱的提取,含量测定采用Agilent XDB-C_(18)(1.8μm,2.1×100 mm)色谱柱,乙腈水溶液梯度洗脱,检测波长278 nm,流速0.3 mL/min,柱温30℃。结果:确定牛大力中刺桐碱80℃水浴回流提取的最佳条件为料液比为1∶15 g/mL,提取时间为1.5 h,乙醇浓度为70%;建立的刺桐碱含量测定方法快速、准确、稳定、可靠。结论:22个不同的牛大力样品中刺桐碱含量差别较大,与品种、生长环境和生长年限等因素有关。

岭南道地药材牛大力质量标准研究进展

牛大力为豆科植物美丽崖豆藤的干燥根,具有补虚润肺、强筋活络之效,为岭南地区著名的药食两用植物。综述了牛大力质量标准收载情况、品种鉴别、检查、有效成分测定方法研究及指纹图谱质量评价等方面的研究进展,分析了已有牛大力质量研究方法的优势与不足,提出了建立牛大力质量综合评价体系的方法,为牛大力药材质量评价研究与质量控制提供参考。

HPLC法同时测定牛大力中豆甾醇和β-谷甾醇含量

【目的】建立一种同时测定牛大力中豆甾醇和β-谷甾醇含量的高效液相色谱法。【方法】采取超声提取法,提取溶剂为100%甲醇,提取料液比为1∶20 g/mL,提取时间为60 min;检测色谱柱为CAPCELL PAK C18(4.6 mm×250 mm,5μm),流动相为甲醇-水(97∶3),流速为1.0 mL/min,柱温为35℃,检测波长为210 nm,进样量为20μL。【结果】豆甾醇、β-谷甾醇分别在0.0202~1.0120 mg/mL(r=0.9997)、0.0202~1.0075 mg/mL(r=0.9998)范围内与峰面积呈现良好的线性关系,平均回收率分别为98.3%、96.8%,RSD分别为1.20%、1.64%,方法精密度、重复性、稳定性均良好。【结论】该方法简便快速、准确稳定、灵敏度高、专属性强,适用于牛大力中豆甾醇和β-谷甾醇含量的测定;该方法可为牛大力进行质量控制提供参考方法,可为牛大力进行深入开发提供科学借鉴。

广东省牛大力茎叶定性质量标准研究

建立广东省牛大力茎叶药材的定性质量标准。收集广东省5个地区牛大力茎叶,按照《中华人民共和国药典》(2020年版)四部通则方法,对其粉末进行显微结构和薄层色谱(TLC)鉴别;用烘干法测定水分,用马弗炉测定总灰分和酸不溶性灰分,采用浸泡法和加热回流法测定浸出物(水溶性、醇溶性)含量,采用电感耦合等离子体发射光谱仪(ICP-OES)测定铅(Pb)、铜(Cu)、砷(As)、镉(Cd)、汞(Hg)等元素含量。结果显示,不同产地牛大力茎叶样品显微特征基本一致,薄层色谱重现性好、专属性强,水分含量为6.83%~9.16%,总灰分含量为3.74%~5.90%,酸不溶性灰分含量为0.49%~0.92%,冷水浸出物含量为3.01%~10.78%,热水浸出物含量为5.88%~15.03%,冷醇浸出物含量为2.38%~4.22%,热醇浸出物含量为3.29%~6.36%,Pb含量为0.185~2.863 mg/kg,Cu含量为1.840~6.601 mg/kg,As含量为未检出~0.028 mg/kg,Cd含量为0.016~0.086 mg/kg,Hg含量为未检出~0.019 mg/kg。本试验方法操作简单,可行性好,结果可靠,可为牛大力茎叶药材定性质量标准的建立提供参考。

蒸制次数对牛大力黄酮和多糖含量的影响

目的研究不同蒸制次数对牛大力饮片的黄酮和多糖含量的影响,为牛大力的蒸制研究提供参考。方法将同一批次牛大力根饮片随机分为对照组和1~5次蒸制组,共6组;分别采用超声波提取法和水提醇沉法提取各组饮片的黄酮和多糖,分别以芦丁和无水葡萄糖作为对照品,通过分光光度计测量黄酮和多糖的含量。结果多次蒸制可提高牛大力总黄酮的析出含量,2~5次蒸制析出含量显著提高,差异具有统计学意义(P<0.05),其中蒸晒3次含量最高;蒸制可提高牛大力多糖的析出含量,但差异无统计学意义(P>0.05)。结论蒸制能显著提高牛大力的总黄酮析出含量,对多糖含量的提高并不显著。

牛大力多糖对育肥期海南猪生长性能、血清生化指标、免疫指标及肠道菌群的影响

试验旨在研究牛大力多糖对育肥期海南猪生长性能、血清生化指标、免疫指标及肠道菌群的影响。选取体重(68.11±1.20)kg、健康、阉割的育肥期海南猪32头,随机分为2组,每组4个重复,每个重复4头猪。对照组猪饲喂基础日粮,牛大力多糖组在基础日粮中添加0.1%牛大力多糖。预试期7 d,试验期70 d。结果显示,与对照组相比,牛大力多糖组海南猪的平均日采食量(ADFI)显著降低(P<0.05);猪血清甘油三酯(TG)含量和谷丙转氨酶(ALT)活性显著降低(P<0.05),高密度脂蛋白胆固醇(HDL-C)含量显著提高(P<0.05);血清免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、免疫球蛋白A(IgA)和肿瘤坏死因子-α(TNF-α)的含量均显著提高(P<0.05)。与对照组相比,牛大力多糖组Chao1物种的丰富度显著降低(P<0.05);在门水平上,脱硫菌门相对丰度显著降低(P<0.05);在科水平上,普雷沃菌科相对丰度显著升高(P<0.05),Muribaculaceae、肠杆菌科、脱硫弧菌科的相对丰度显著降低(P<0.05);在属水平上,丁酸厌氧杆菌、高氏瘤胃球菌群(Ruminococcus_gauvreauii_group)、粪球菌属、月形单胞菌属、布劳特氏菌属的相对丰度显著上调(P<0.05),大肠志贺氏杆菌、脱硫弧菌、Shuttleworthia的相对丰度显著下调(P<0.05)。HDL-C含量与Marvinbryantia的相对丰度呈显著正相关(P<0.05)。研究表明,日粮中添加0.1%牛大力多糖能够降低育肥期海南猪的ADFI,改善血脂代谢,提高机体免疫力,优化结肠菌群结构,促进有益菌生长和抑制有害菌增殖。

牛大力提取物对小鼠的急性毒性与遗传毒性研究

本研究评价了牛大力提取物的急性毒性和遗传毒性。采用限量法进行急性毒性试验;采用平板掺入法,对有组氨酸营养缺陷的5株鼠伤寒沙门氏菌突变型菌株TA97a、TA98、TA100、TA102和TA1535进行回变菌落计数;经口灌胃法进行哺乳动物红细胞微核试验,计数骨髓嗜多染红细胞占总红细胞的比例及含微核嗜多染红细胞比率;采用中国仓鼠肺细胞(CHL)培养技术进行染色体畸变试验,观察受试样品牛大力提取物致CHL细胞株染色体数目及结构变化。结果显示,受试样品牛大力提取物属实际无毒级物质,细菌回复突变试验、红细胞微核试验、染色体畸变试验结果均为阴性。牛大力提取物在本试验剂量范围内未观察到急性毒性和遗传毒性。

牛大力水提物对斑马鱼抗氧化和免疫能力的影响

为探究牛大力(Millettia speciosa Champ.,MSP)水提物对斑马鱼(Danio rerio)抗氧化和免疫能力的影响,本研究在斑马鱼养殖水体中加入4个不同水平(5、15、25和35 mg/L)的MSP水提物分别饲养,14 d后分析斑马鱼肠道、肝脏的抗氧化能力和免疫指标,并使用嗜水气单胞菌(Aeromonas hydrophila)进行攻毒试验,统计攻毒后各组斑马鱼的死亡率。结果表明,MSP水提物25 mg/L组斑马鱼肠道和肝脏的超氧化物歧化酶(Superoxide Dismutase,SOD)、溶菌酶(Lysozyme,LZM)的活性和总抗氧化能力(Total Antioxidant Capacity,T-AOC)均显著高于空白对照组,同时丙二醛(Malondialdehyde,MDA)含量明显降低;与免疫相关的防御素(Defensins)、肿瘤坏死因子α(Tumor necrosis factorα)、核转录因子κB(Nuclear factorκB)、白细胞介素-1(Interleukin-1)、溶菌酶和补体(Complement)等基因(分别简写为defbl1、tnf-α、nfκb、il-1β、lyz、c3a)的相对表达量也显著高于空白对照组(P<0.05)。嗜水气单胞菌感染结果显示,试验组的存活率均高于空白对照组,其中35 mg/L组免疫保护率达到最高,为28.6%。由此可知MSP水提物可以有效提高斑马鱼的抗氧化和免疫能力,增强其机体的免疫力。

牛大力总黄酮提取工艺优化及其抗氧化活性研究

采用超声波辅助法提取牛大力总黄酮。在单因素试验的基础上,通过响应面法优化总黄酮的提取工艺,并评价其抗氧化活性。结果表明:最佳提取工艺为乙醇体积分数59%、液料比53∶1 (mL/g)、提取时间50 min、提取温度67℃,在此条件下总黄酮提取量为3.357 mg/g。牛大力总黄酮对ABTS、OH自由基的IC_(50)分别为16、88μg/mL,具有较强的抗氧化活性。

广西牛大力品质研究

【目的】全面分析与评价广西牛大力品质,挖掘广西牛大力质量优势,助力广西地方特色资源优势转化为产业优势。【方法】实验根据质量控制要求,对广西不同地区牛大力中的水分、浸出物、多糖、总黄酮、总甾醇、总酚酸、金属元素、农药残留量等质量指标和卫生指标进行测定。【结果】测定结果表明广西牛大力水分含量均值为59.2 g/100 g,水冷浸出物含量均值为13.2%,水热浸出物含量均值为28.9%,醇冷浸出物含量均值为12.7%,醇热浸出物含量均值为13.5%,多糖含量均值为12.63%,总黄酮含量均值为48.79 mg/g,总甾醇含量均值为22.44 mg/g,总酚酸含量均值为35.05 mg/g,Mn、Fe、Zn、Se、As、Cd和Pb等7种金属含量有所差异,吡虫啉、毒死蜱和乐果等19种农药均未检出。【结论】本研究提供了广西牛大力水分、浸出物、多糖、总黄酮、总甾醇、总酚酸、金属元素、农药残留等含量范围,为指导牛大力新品种选育、种植栽培、初加工、精深加工产品开发、牛大力及其精深加工产品的质量控制提供具有一定科学价值的参考。

不同栽培措施对牛大力产量的影响

为满足牛大力产业化发展的需求,提高牛大力的产量,本研究以牛大力营养杯苗为材料,研究不同种植地、整地方法、种植密度等栽培措施对牛大力产量的影响。结果表明:种植在广西桂林兴安、桂林全州两地的牛大力产量为60 000 kg/ha及以下,种植在广西钦州灵山、南宁上林、百色田林以及广东湛江的牛大力产量为60 000 kg/ha以上;3种整地方法中,单株平均产量最高的是全垦,为3.31 kg;种植密度为15 000株/ha时,药材产量最高;与对照组相比,4个不同基肥处理组产量都明显增加,其中生物有机肥(1 300 kg/ha)+复合肥(200 kg/ha)>生物有机肥(1 500 kg/ha)>沼肥(1 200 kg/ha)>普通化肥(尿素75 kg/ha,磷酸二胺300 kg/ha,过磷酸钙225 kg/ha)>对照(不施肥);随着种植年限的延长,牛大力的产量越高,品质越好,但种植成本也会增加。综合考量,牛大力最佳整地方式为全垦,最佳种植密度为15 000株/ha,种植地以北回归线以南的地区最佳,基肥以生物有机肥+复合肥最优,种植年限以4年最宜。

响应面法优化牛大力总黄酮提取工艺研究

[目的]对牛大力中总黄酮的提取工艺进行优化,建立其含量测定方法。[方法]此研究以市售广西牛大力饮片为对象,在单因素实验基础上,筛选出影响牛大力总黄酮提取量较显著的液料比、超声时间和乙醇浓度三个因素,采用Box-Behnken软件设计响应面试验优化牛大力中总黄酮提取工艺条件。[结果]结果表明,牛大力总黄酮提取的最优工艺条件为液料比43.04∶1(mL/g),超声时间37.05 min,乙醇浓度36.89%,预测总黄酮提取量为39.3286mg/g。[结论]此提取工艺的研究可为牛大力的开发利用提供参考。