Estimation of boswellic acids in herbal formulations containing Boswellia serrata extract and comprehensive characterization of secondary metabolites using UPLC-Q-Tof-MS^e

Boswellia serrata is a widely used herb in Indian systems of medicine and is well known for its potential medicinal properties.A chromatographic method was developed for the analysis and quanti cation of six boswellic acid marker compounds,i.e.,keto boswellic acid(1),3-O-Acetyl 11-keto-boswellic acid(2),-b Boswellic acid(3),-Boswellic acid(4),3-O-Acetyl--boswellic acid(5)and 3-O-Acetyl--boswellic acid b(6)in commercial herbal products containing B.serrata as an ingredient.Combining UPLCbwith Q-Tof-MS/MS makes the better identi cation of secondary metabolites and adulterants in the herbal formulations containing B.serrata in rapid time using fragmentation approach than the traditional approaches.In this study quanti cation of boswellic acids with UPLC-PDA method was performed as per the pharmacopeia guidelines.Furthermore,minor phytochemical constituenBoswellia serrata is a widely used herb in Indian systems of medicine and is well known for its potential medicinal properties.A chromatographic method was developed for the analysis and quantification of six boswellic acid marker compounds,i.e.,keto boswellic acid(1),3-O-Acetyl 11-keto b-boswellic acid(2),ɑ-Boswellic acid(3),b-Boswellic acid(4),3-O-Acetyl-ɑ-boswellic acid(5)and 3-O-Acetyl-b-boswellic acid(6)in commercial herbal products containing B.serrata as an ingredient.Combining UPLC with Q-Tof-MS/MS makes the better identification of secondary metabolites and adulterants in the herbal formulations containing B.serrata in rapid time using fragmentation approach than the traditional approaches.In this study quantification of boswellic acids with UPLC-PDA method was performed as per the pharmacopeia guidelines.Furthermore,minor phytochemical constituents were identified and characterized with the help of LC-Q-Tof-MS/MS fragmentation data and various isoforms of boswellic acids and tirucallic acids in B.serrata oleo-gum-resin extract were identified.ts were identi ed and characterized with the help of LC-Q-Tof-MS/MS fragmentation data and various isoforms of boswellic acids and tirucallic acids in B.serrata oleo-gum-resin extract were identi ed.

Chemical composition and anti-inflammatory activity of Boswellia ovalifoliolata essential oils from leaf and bark

We isolated the essential oils from Boswellia ovalifoliolata N.P.Balakr and A.N.Henry using hydrodistillation,identified the obtained compounds using gas chromatography–mass spectrometry(GC–MS) and Fourier transform-infrared spectroscopy(FT-IR),and studied the photophysical and electrochemical properties using UV–visible and fluorescence spectroscopy and cyclic voltammetry.On the basis of GC–MS spectra,38 compounds were identified in the essential oil from leaves and 26 in the oil from bark.The bark oil contained 13.44% b-Farnesene(sesquiterpene),10.45% caryophyllene oxide(sesquiterpene) and 6.6% spathulenol,(2 Z,6 E)-(sesquiterpene alcohol); the leaf oil contained 11.1% spathulenol(sesquiterpene alcohol),9.0% caryophyllene oxide(sesquiterpenoids) and 6.3% decyl acetate(ester).FT-IR spectra confirmed the presence of aliphatic aldehydes and ketones,carboxylic acid,alcohols,esters and ethers in both oil types.UV–visible absorption spectra showed maximum absorbance at 245,290 and 402 nm for bark oil,and 250,285 and 325 nm for leaf.Bark oil showed strong emission with maximum emission wavelength at 456 nm was higher than that of leaf oil at 414 nm.The essential oil had significant anti-inflammatory activity,and the reduction potential of the leaf oil was-0.44 and-0.56 e V for bark.

Boswellia resin extract and propolis derived polyphenols in patients with type III chronic prostatitis/chronic pelvic pain syndrome:An Italian prospective multicenter study

Objective:To assess the efficacy and safety of a treatment regimen based on rectal administration of Boswellia resin extract and propolis derived polyphenols in patients with type IIIa and type IIIb chronic prostatitis and chronic pelvic pain syndrome(CP/CPPS).Methods:Patients with type IIIa and type IIIb CP/CPPS received one rectal suppository a day for 15 days per month for 3 consecutive months.Participants were evaluated with National Institutes of Health-Chronic Prostatitis Symptom Index(NIH-CPSI),the International Prostate Symptom Scores(IPSS),International Index of Erectile Function(IIEF),four-glass test,uroflowmetry,and prostate-specific antigen assessments at baseline and at Week 4,and Week 12.Primary endpoints were improvement in pain domain of NIH-CPSI and improvement of NIH-CPSI total score.Secondary outcomes included improvement of micturition and quality of life(QoL)domains of NIH-CPSI questionnaire.Results:A total of 61 males were enrolled.No adverse events were reported.Significant improvements from baseline to Day 30 were reported for NIH-CPSI total score(mean difference:-9.2;p<0.01),NIH-CPSI pain domain(mean difference:-5.5;p<0.01),NIH-CPSI micturition domain,NIH-CPSI QoL domain,and IPSS total score(mean difference:-5.6;p<0.01).No significant changes from baseline in terms of IIEF score or maximum flow rate were observed.At final follow-up(Day 90),further significant improvements in terms of NIH-CPSI total score(mean difference:-12.2;p<0.01),NIH-CPSI pain domain(mean difference:-6.6;p<0.01),NIH-CPSI micturition domain,NIH-CPSI QoL domain,and IPSS total score were reported.Conclusion:Rectal administration of Boswellia resin extract and propolis derived polyphenols is well tolerated and delivers a significant symptomatic improvement in most patients with type IIIa and type IIIb CP/CPPS.

Parasitic plant in natural Boswellia papyrifera stands at Humera, Northern Ethiopia

In Boswellia papyrifera （Del.） Hochst natural stands, we stud- ied the association of parasitic plants with B. papyrifera trees from which frankincense was tapped and marketed for domestic and export markets. Data on the rate of infection of parasitic plants on B. papyrifera was collected in three transects located at separate locations around Baha kar, northern Ethiopia. Each transect had ten circular sample plots of 400 m2 and separated by 100 m. Species composition, DBH, height, crown di- ameter, number of main, secondary and tertiary branches and number of parasitic plants on individual trees were recorded. Sixteen tree species were recorded in the combined sample plots. The parasitic plant associ- ated with B. papyrifera was identified as Tapinanthus globiferus. This parasite infected 38% of Boswellia trees in sample plots. The infection rate of the parasitic plant varied from 1 to 33 per Boswellia tree. The infection of T. globiferus on B. papyrifera was predominantly limited to tertiary small branchlets arising from secondary branches; parasitic plants were absent on thick main and secondary branches. In all plots, infectionof T. globiferus was exclusively limited to Boswellia trees. The influence of T. globiferus parasitism on growth of Boswellia trees and its influence on yield of incense production needs further investigation. Management of natural stands for frankincense production should include measures to reduce infection by T. globiferus.

Gum resin extract of Boswellia serrata attenuates lipopolysaccharide-induced inflammation and oxidative damage in hepatic and renal tissues of rats

Objective:To explore the effect of ethyl acetate gum resin extract of Boswellia serrata on lipopolysaccharide(LPS)induced inflammation and oxidative damage in hepatic and renal tissues of rats.Methods:The rats were divided into four groups:control,LPS,LPS+Boswellia serrata extracts(100 mg/kg and 200 mg/kg).LPS(1 mg/kg)and the extract(100 and 200 mg/kg,30 min before LPS)were administered intraperitoneally for 3 weeks.The levels of liver enzymes,albumin,total protein,creatinine,blood urea nitrogen(BUN),interleukin(IL)-6,malondialdehyde(MDA),and total thiol groups and superoxide dismutase(SOD)and catalase(CAT)activities were measured.Results:The levels of liver enzymes,creatinine,and BUN,IL-6,MDA in the LPS group were markedly increased(P<0.001)while albumin,total protein,and total thiol concentration,as well as SOD and CAT activities,were decreased compared with the control group(P<0.05 or 0.01).Boswellia serrata extracts diminished the levels of liver enzymes,creatinine,BUN,IL-6,and MDA(P<0.01 and P<0.001),and elevated the concentration of total protein and total thiol and SOD and CAT activities(P<0.05 or 0.01).Conclusions:The ethyl acetate gum resin extract of Boswellia serrata reduces LPS-induced inflammatory reactions and oxidative damage,thus ameliorating hepatic and renal function.

Antioxidant and hepatoprotective effects of Boswellia ovalifoliolata bark extracts

Paracetamol(PCM) hepatotoxicity is related to reactive oxygen species(ROS) formation and excessive oxidative stress; natural antioxidant compounds have been tested as an alternative therapy. This study evaluated the hepatoprotective activity of an alcoholic extract of Boswellia ovalifoliolata(BO) bark against PCM-induced hepatotoxicity. BO extract also demonstrated antioxidant activity in vitro, as well as scavenger activity against 2, 2-diphenyl-1-picrylhydrazyl. Administration of PCM caused a significant increase in the release of transaminases, alkaline phosphatase, and lactate dehydrogenase in serum. Significant enhancement in hepatic lipid peroxidation and marked depletion in reduced glutathione were observed after parac intoxication with severe alterations in liver histology. BO treatment was able to mitigate hepatic damage induced by acute intoxication of PCM and showed a pronounced protective effect against lipid peroxidation, deviated serum enzymatic variables, and maintained glutathione status toward control. The results clearly demonstrate the hepatoprotective effect of BO against the toxicity induced by PCM.

Isolation of Boswelliains A-E,Cembrane-Type Diterpenoids from Boswellia papyifera,and an Evaluation of Their Wound Healing Properties

Boswelliai ns A-E(1-5),five new cembra ne-type diterpenoids,were isolated from the resin of Boswellia papyifera.Their structures,determined by using spectroscopic,computational and chemical methods,demonstrate that 2-5 are novel 4,5-seco-,4,5-seco-,9,10-seco-,and 6,7-seco-cembrane diterpenoids,and that 1,4 and 5 bear aldehyde moieties.An evaluation of the wound healing properties of these natural products along with a derivative of 2 shows that compounds 2 and 3 significantly stimulate proliferation,mobility and tuber formation of human umbilical vein endothelial cells(HUVECs)at the concentration of 20μmol/L.

乳香提取物调控Wnt/β-catenin信号通路抑制结直肠癌细胞侵袭转移的作用机制研究

目的研究乳香提取物对人结肠癌细胞HCT-116增殖和侵袭转移的影响及可能的作用机制。方法体外培养人结肠癌细胞珠HCT-116,使用不同浓度乳香提取物乙酰基-11-酮基-β-乳香酸(AKBA)分别作用于细胞相同时间后,以CCK-8实验、Transwell细胞侵袭实验、划痕实验检测其对细胞增殖和侵袭转移的影响,以Western blotting、荧光定量PCR等方法探究其对细胞侵袭转移相关蛋白、mRNA表达的影响。结果AKBA可抑制人结肠癌细胞HCT-116的增殖、迁移能力,且呈剂量依赖(P<0.05);AKBA可抑制C-Myc、β-catenin的蛋白和mRNA表达,且呈剂量依赖(P<0.05)。结论AKBA可以抑制人结肠癌细胞HCT-116的增殖和迁移,诱导细胞凋亡。这可能与其调控Wnt/β-catenin信号通路相关。

New tirucallane-type triterpenoids from the resin of Boswellia carterii and their NO inhibitory activities

Six new tirucallane-type triterpenoids(1-6),along with ten known triterpenoids,were isolated from methylene chloride extract of the resin of Boswellia carterii Birdw.By the application of the comprehensive spectroscopic data,the structures of the compounds were clarified.The experimental electronic circular dichroism spectra were compared with those calculated,which allowed to assign the absolute configurations.Compounds 5 and 6 possesed a 2,3-seco tirucallane-type triterpenoid skeleton,which were first reported.Their inhibitory activity against NO formation in LPS-activated BV-2 cells were evaluated.Compound 9 showed appreciable inhibitory effect,with an IC_(50)value of 7.58±0.87μmol·L^(-1).

Cardioprotective and antioxidant effects of oleogum resin "Olibanum" from Boswellia carteri Birdw.(Bursearceae)

One of the leading causes of death worldwide is cardiovascular disease, hence searching for a cure is an important endeavor. The totally safe, edible, and inexpensive Boswellia plant exudate, known as olibanum or frankincense, is considered to possess diverse medicinal values in traditional medicine and from recent biological studies. Investigating the cardioprotective and antioxidant activities of olibanum from a Boswellia species, family Bursearaceae, namely Boswellia carteri Birdw. was the aim of this study. Cardioprotective activity was evaluated using a model of myocardial infarction induced by isoprenaline(ISO), while antioxidant activity was tested adopting nitric oxide scavenging(NOS) and azino-bis-3-ethyl benzthiazoline-6-sulfonic acid(ABTS) assays. The results revealed a mild cardioprotective effect and weak antioxidant activity.

Plants Extracts and a Mycoinsecticide in Cowpea Yield Improvement in Guinean Savanah and Sahelian Savanah Agro-Ecological Zones of Cameroun

Cowpea yield improvement is done by adding agricultural inputs. The use of natural substances as pesticides is being encouraged to fight against cowpea field pests. The pesticidal potentials of Azadirachta indica and Boswellia dalzielii water extracts, Metarhizium anisopliae, alone and in combination with plant extracts, and a commercial synthetic pesticide (Decis®) were tested in field on two varieties of cowpeas, Vigna unguiculata in two agroecological zones (Guinean Savanah and Sahelian Savanah) of Cameroon. The field trials were carried out in a full randomized block design including nine treatments and a control. Four replications were made concerning the different treatments and control. At the flowering stage, the cowpea field was sprayed three times with different pesticidal formulations at the interval of five days. The number of ramifications per plant, and that of pods per block and seed yield were determined. The pesticide formulations considerably (p ®). Bafia variety treated with the combination of M. anisopliae and A. indica recorded the highest ramification rate (37.03 ± 1.59) in Maroua (Sahelian Savanah). The same variety also produced more important pods number (90.50 ± 16.66) in Ngaoundere (Guinean Savanah) with the binary combination of two plants used in this experiment. The highest seed yield (44.23 ± 2.31) was recorded in Ngaoundere with B125 variety treated with the combination of the three pesticidal formulations separately (A. indica, B. dalzielii, M. anisopliae). The plant products used in this work, M. anisopliae and their mixtures could supersede the synthetic pesticides considering environmental issue in cowpea crop protection. Then, such formulations would not only improve crop productivity but also preserve environment from the pollution due to the use of synthetic residual chemicals.

乳香挥发油抑制人肝癌SMMC-7721细胞株增殖及诱导凋亡的作用

目的探讨乳香挥发油体外抑制人肝癌SMMC-7721细胞株增殖及诱导凋亡的作用。方法使用四甲基偶氮唑蓝(MTT)法观察乳香挥发油对SMMC-7721的增殖抑制作用;通过吖啶橙染色,观察SMMC-7721的形态学变化;使用琼脂糖凝胶电泳法检测乳香挥发油对细胞DNA降解的影响;利用流式细胞术分析乳香挥发油诱导SMMC-7721凋亡的凋亡率及对细胞周期的影响;应用间接免疫荧光法观察凋亡调控基因bax和bcl-2蛋白的表达变化。结果乳香挥发油浓度为0.7、0.07、0.007、0.0007、0.00007mg.mL-1作用24h时,能抑制SMMC-7721的生长,并呈浓度依赖性。在浓度为0.07mg.mL-1时,作用48h或72h,电泳结果显示了DNA梯状条带,流式细胞仪检测到凋亡峰,且细胞被阻滞于G1期。间接免疫荧光法的结果表明,促凋亡蛋白bax的表达增强,抗凋亡蛋白bcl-2的表达无明显变化。结论乳香挥发油能抑制肝癌细胞株SMMC-7721的增殖,并可能通过上调线粒体内bax/bcl-2的表达比例诱导SMMC-7721细胞的凋亡,而且其诱导的凋亡具有细胞周期依赖性。

乳香抗哮喘有效部位化学成分的研究

目的研究乳香抗哮喘活性部位的化学成分。方法以抗哮喘活性为追踪,采用硅胶柱色谱对乳香抗哮喘有效部位进行分离,运用IR、MS、NMR等方法进行结构鉴定。结果从乳香的抗哮喘活性部位分离并鉴定了8个化合物,分别为β乙酰乳香酸(acetyl β-boswellic acid,Ⅰ)、β-乳香酸(B-boswellic acid,Ⅱ)、pinnicolic acid A(3-ox-olanosta-8,24-dien-21-oic acid,Ⅲ)、栓菌酸β(3β-hydroxylanosta-8,24-dien-21-oic acid,Ⅳ)、α-乳香酸(α-boswellic acid,Ⅴ)、11-羰基-β-乙酰乳香酸(11-keto-β-boswellic acid,Ⅵ)、脱氢松香酸(Ⅶ)、山达海松酸(Ⅷ)。结论化合物Ⅶ、Ⅷ均为首次从该植物中分离得到。

大豆甙元(S86019)与乳香有效成分Bc-4或阿糖胞苷对HL-60细胞分化的联合诱导

大豆甙元($86019)单独处理HL-60细胞,对细胞生长诱导分化作用较弱,NBT阳性细胞数低于10%。当S86019与乳香有效成分Bc-4联合应用时,对HL-60细胞的生长有明显抑制,并有明显的分化诱导;S86019与Bc-4或Ara-c联合应用,对细胞的增殖有较强的抑制和分化诱导作用。S86019与Bc-4联合应用4d,NBT阳性细胞达80%,75%细胞获得吞噬乳胶颗粒能力,90%细胞向成熟粒细胞方向分化;S86019与Ara-c联合应用4d,NBT阳性反应细胞达70%,82%细胞获得吞噬能力,90%细胞形态发生分化。S86019与Bc-4或Ara-c联合用药4d,可明显阻断细胞由G1期向s期移行,G1期细胞数明显增加。

乳香的化学成分

目的 研究乳香的化学成分。方法 应用色谱技术进行分离和纯化 ,IR ,NMR ,MS等波谱解析化学结构。结果 从乳香中分离出 6个化合物 ,分别鉴定为α 乙酰乳香酸 (1) ,β 乙酰乳香酸 (2 ) ,羽扇 2 0 (2 9) 烯 3α 乙酰氧基 2 4 酸 (3) ,α 乳香酸 (4) ,β 乳香酸 (5 ) ,11 羰基 β 乙酰乳香酸 (6 )。 结论 化合物 3为新成分。

HPLC法测定乳香中乙酰乳香酸类成分的含量

目的:建立乳香中3种乙酰乳香酸类成分的HPLC含量测定方法,为全面评价乳香药材质量提供依据。方法:采用HPLC方法测定,Diamonsil(钻石)C8色谱柱(250 mm×4.6mm,5μm);乙腈-0.1%磷酸(A:80:20;B:70∶30)为流动相;检测波长210nm,250nm;柱温35℃。结果:α-乙酰乳香酸、β-乙酰乳香酸、11-羰基-β-乙酰乳香酸分别在0.099～9.9μg(0.999 9)、0.097 6～9.76μg(0.999 9)、0.099 7～9.97μg(0.999 9)范围内呈线性,平均回收率分别为98.6%、101.9%、98.25%,RSD分别为1.38%、1.49%、1.86%(n=6);乳香药材供试品中3种乙酰乳香酸的百分含量分别在0.23%～2.41%、0.45%～4.27%、0.35%～5.27%范围。结论:本方法简便,稳定,可用于乙酰乳香酸类成分的定量分析。

气质联用法分析炮制对乳香挥发油的影响

研究了乳香炮制前后挥发油含量的变化，用ＧＣ－ＭＳ法分析鉴定了挥发油中的组分，并用ＧＣ法测定了各种组分炮制前后的含量变化。结果表明，乳香经炮制后各炮制品的挥发油含量变化不大，其组分含量有一定变化。

乳香提取物固体纳米脂质微粒的制备与性能研究

采用高剪切超声乳化法制备乳香提取物固体纳米脂质微粒(BCBE-SLN),通过改变剪切速率、乳化温度和配方组成,得出适宜的制备条件;对所得产品的粒径、Zeta电位及形貌进行了表征;研究了其离心、储存及稀释稳定性;并对其进行体外经皮试验。结果表明:在乳化温度为60℃,乳化剂鲸蜡醇聚醚-20(BC-20)和硬脂醇聚氧乙烯醚-25(A-25)的质量分数均为0.5%,脂质原料和乳香提取物的质量分数均为1.0%,剪切速率为13 000 r/min,剪切时间为3 min时,制备的BCBE-SLN的平均粒径为70.6 nm,多分散指数(PDI)为0.388,Zeta电位为-20.0 m V,产品有很好的皮肤渗透性,并具备良好的离心、储存及稀释稳定性。

萃取—紫外法测定乳香挥发油脂质体的包封率

目的建立测定乳香挥发油（BCBV）脂质体包封率的方法。方法采用石油醚萃取-紫外法测定乳香挥发油脂质体的包封率。结果乳香挥发油在307nm附近有最大吸收峰，浓度在41．36～310．20μg·ml^-1范围内与吸光度A呈良好的线性关系，精密度良好（RSD=0．3％），低、中、高三个浓度的乳香挥发油溶液平均回收率均符合规定，分别为98．5％、99．3％、98．1％。结论各项方法学考察指标均符合规定，适用于乳香挥发油脂质体包封率的测定。

乳香诱导急性早幼粒细胞白血病细胞凋亡与细胞周期改变

目的 :为探讨乳香抗白血病的作用机制 ,研究乳香诱导急性早幼粒细胞白血病细胞凋亡与细胞周期关系。方法 :应用形态学观察 ,DNA凝胶电泳、DNA片段百分率 ,流式细胞仪分析检测白血病细胞凋亡及细胞周期。结果 :乳香提取物与全反式维甲酸相似 ,具有诱导急性早幼粒细胞白血病细胞凋亡及作用细胞周期 G1 及 S期( G1 期细胞减少 ,S期细胞增多 )。结论