RP-HPLC法测定辣椒中辣椒素、二氢辣椒素和降二氢辣椒素含量

目的:建立反相高效液相色谱法同时测定辣椒中辣椒素、二氢辣椒素和降二氢辣椒素含量。方法:采用Nucleosil C18(4.6 mm×250 mm,5μm)色谱柱,流动相为乙腈-水,梯度洗脱,流速1.0 mL.min-1,检测波长280 nm,柱温45℃,外标法定量。结果:3个辣椒碱的线性关系良好,线性范围均为1.0～160.0μg.mL-1,相关系数在0.9991～0.9995之间;精密度、稳定性试验的RSD均低于3%;辣椒素、二氢辣椒素和降二氢辣椒素平均回收率(n=6)分别为96.4%,98.1%,99.3%,RSD分别为2.6%,2.1%,1.2%;检出限分别为0.2,0.1,0.1μg.mL-1。结论:该方法简便快捷,应用本方法测定了10个品种辣椒中3个辣椒碱的含量,结果稳定。     

一测多评法测定藏药小米辣中3种辣椒碱的含量

目的：建立藏药小米辣中3种辣椒碱类化学成分含量测定的一测多评法,探讨一测多评法在民族药质量控制中的应用。方法：采用超高压液相色谱法,使用Waters ACQUITY BEH C18（2.1 mm×100mm,1.7 μm）色谱柱,流动相为乙腈-水（50︰50）,流速：0.2 mL·min^-1,柱温30℃,检测波长280 nm,进样量1 μL。以辣椒素为内参物,分别建立二氢辣椒素、降二氢辣椒素相对于辣椒素的相对校正因子,分别采用外标法和一测多评法测定3种辣椒碱的含量,并通过相对标准偏差（RSD）和Pearson系数比较二者结果的相对误差。结果：降二氢辣椒素、辣椒素、二氢辣椒素进样量分别在2.21~22.1 ng、8.96~89.6 ng、3.75~37.5 ng范围内线性关系良好;降二氢辣椒素和二氢辣椒素相对于辣椒素的相对校正因子分别为0.970和1.159;且在不同实验条件下重现性良好（RSD < 3.0%）;一测多评法的计算结果与外标法测得结果无显著差异。10批不同产地小米辣样品中降二氢辣椒素、辣椒素和二氢辣椒素的含量范围分别为0.0281%~0.0534%、0.258%~0.389%和0.115%~0.209%。结论：建立的一测多评法可作为小米辣中3种辣椒碱类化学成分的含量测定方法,一测多评法为民族药质量控制提供了一个新的模式与方法。     

HPLC法测定辣椒秸秆中辣椒素和二氢辣椒素含量

目的:检测分析超越馨香和福椒六号辣椒品种秸秆不同时期叶片、侧枝、主杆部位的辣椒素与二氢辣椒素含量,为有效利用辣椒秸秆提供依据。方法:辣椒秸秆分成叶片、侧枝、主杆,60℃烘干粉碎,与甲醇溶液充分混合,采用HPLC法测定辣椒素和二氢辣椒素含量,并用精确度、浓缩、重复性、稳定性和加样回收率试验检验测定方法的可靠性。HPLC色谱条件:采用Agilent C18(4.6 mm×250 mm,5μm)色谱柱,以甲醇-超纯水(85:15)为流动相,流速1 mL·min-1,检测波长280 nm,柱温37℃。结果:辣椒素与二氢辣椒素的标准和浓缩曲线呈显著的线性变化;连续5次进样,RSD分别为0.13%和0.08%;重复性试验,样品辣椒素和二氢辣椒含量的RSD分别为1.7%和2.7%;稳定性试验,12 h内供试品溶液中辣椒素和二氢辣椒素浓度的RSD分别为2.3%和1.7%;回收率分别为95.8%和94.3%。除超越馨香,辣椒素和二氢辣椒素含量在不同收获时间和部位之间均存在显著差异,并存在时间和部位的交互效应,叶片辣椒素和二氢辣椒素含量显著高于侧枝和主杆(P<0.05)。结论:本研究采用HPLC法检测辣椒秸秆中辣椒素和二氢辣椒素含量的方法是可行的;辣椒秸秆中含有较高浓度的辣椒素和二氢辣椒素,其含量与品种、部位和采收时间有关。     

高效液相色谱-串联质谱法测定羌活中异欧前胡素和紫花前胡苷北大核心CSCD

目的:建立高效液相色谱-串联质谱法测定羌活中异欧前胡素和紫花前胡苷含量。方法:羌活样品超声萃取甲醇稀释后分析。色谱分离采用C18反相色谱柱(150 mm×2.1 mm,3.5μm),流动相为0.1%乙酸水溶液和乙腈,梯度洗脱。串联质谱在多反应监测模式下检测目标分析物,以保留时间和特征离子对(母离子和2个碎片离子)信息比较进行定性和定量分析。结果:异欧前胡素和紫花前胡苷的检出限分别为0.03 ng·mL-1和0.015 ng·mL-1,定量限分别为0.10 ng·mL-1和0.05ng·mL-1,异欧前胡素和紫花前胡苷平均回收率分别为93.6%和94.5%,RSD分别为2.4%和2.6%。结论:该方法经方法学验证,可用于羌活中异欧前胡素和紫花前胡苷的分析。     

HPLC法同时测定关节解痛膏中的辣椒素、二氢辣椒素、欧前胡素和异欧前胡素

目的：建立HPLC测定关节解痛膏中辣椒素、二氢辣椒素、欧前胡素和异欧前胡素含量的方法。方法：CAPCELL PAK C18色谱柱（250 mm×4.6 mm,5μm）;流速：0.8 ml·min^-1;流动相A：甲醇-乙腈（1∶1）,流动相B：1%磷酸溶液,梯度洗脱;检测波长分别为280 nm（辣椒素和二氢辣椒素）和254 nm（欧前胡素和异欧前胡素）。结果：辣椒素、二氢辣椒素、欧前胡素和异欧前胡素分别在0.298-5.956μg（r=0.999 8）、0.152-3.044μg（r=0.999 6）、0.018-0.352μg（r=0.999 5）、0.010-0.204μg（r=0.999 3）范围内进样量与峰面积呈良好的线性关系,平均加样回收率分别为97.8%、97.0%、96.6%、98.1%,RSD分别为1.02%、0.76%、0.65%、1.35%（n=6）。结论：方法简便、准确、灵敏、重复性好,可作为关节解痛膏中辣椒素、二氢辣椒素、欧前胡素和异欧前胡素的含量控制方法。     

液相色谱-串联质谱法测定人血浆氯吡格雷浓度及其生物等效性评价

目的:建立液相色谱-串联质谱法(LC-MS/MS)测定人血浆中氯吡格雷的浓度,研究2种硫酸氢氯吡格雷片的人体药动学及相对生物利用度。方法:血浆样品中加入内标美利曲辛,经乙腈沉淀蛋白提取,采用液相色谱-串联质谱法。用建立的方法测定20例健康男性受试者单剂量口服硫酸氢氯吡格雷受试制剂或参比制剂后的血药浓度,求得药动学参数,并对2种制剂的生物等效性进行评价。结果:在0.02～20 ng·mL-1内呈良好的线性关系,方法回收率98.4%～103.2%,日内、日间RSD均小于15%。单次口服75 mg硫酸氢氯吡格雷受试制剂或参比制剂后的Cmax分别为(1.9±1.5)ng·mL-1和(1.8±1.1)ng·mL-1;tmax分别为(0.8±0.5)h和(1.0±0.8)h;t1/2分别为(3.4±1.6)h和(3.5±1.5)h;AUC(0-48)分别为(4.4±4.3)h·ng·mL-1和(4.4±4.6)h·ng·mL-1;AUC(0-∞)分别为(4.7±4.4)h·ng·mL-1和(4.7±4.7)h·ng·mL-1。受试制剂对参比制剂的相对生物利用度为(98.2±32.8)%。结论:该方法灵敏,无杂质干扰。测得的受试制剂与参比制剂的主要药动学参数之间无明显差异,表明2种制剂在人体内生物等效。     

在线SPE-HPLC系统在补骨脂二氢黄酮甲醚药代动力学研究中的应用

目的:探讨在线固相萃取(SPE)-高效液相色谱(HPLC)系统在小鼠血浆中补骨脂二氢黄酮甲醚药代动力学研究的应用。方法:基于在线SPE-HPLC技术的Ultimate 3000系统测定补骨脂二氢黄酮甲醚血药浓度。色谱条件:采用Venusil MPC18(150 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-5 mmol·L-1磷酸二氢钾缓冲液(pH 3.5),流速1.0 mL·min-1,梯度洗脱;使用Capcell MF Ph-1在线SPE柱(10 mm×4 mm,5μm),洗脱剂为5 mmol·L-1磷酸二氢钾缓冲液(pH 3.5);检测波长238 nm。WinNonlin 5.2软件计算药动学参数。结果:补骨脂二氢黄酮甲醚线性范围为20.0～4000 ng·mL-1,最低定量限为20.0 ng·mL-1,提取回收率为90.5%～94.6%,日内、日间精密度试验的RSD均小于10%,短期稳定性、冻融稳定性及长期稳定性准确度为92.38%～101.7%。结论:该方法工作流程简化,样品分析快速,提高了工作效能,其结果可信且重复性好。     

液相色谱-串联质谱法测定Beagle犬血浆中黄体酮浓度及其药动学

目的:建立液相色谱-串联质谱法(LC-MS/MS)测定Beagle犬血浆中黄体酮含量。方法:取Beagle犬血浆样品,加入适量内标(炔诺酮),用液相萃取技术,经超高效液相色谱分离。色谱柱:Agilent ZORBAX-SB C18分析柱(150 mm×2.1 mm,5μm);保护柱:Agilent C18分析柱(10 mm×2.1 mm,5μm);流动相:甲醇-0.1%甲酸水溶液(90∶10);流速0.2 mL.min-1。采用APCI离子化-串联质谱多离子反应监测(MRM)模式进行检测,放电电流(NC)=3.00,雾化气(GS1)=30.00,辅助气(GS2)=50.00,气帘气(CUR)=30.00,温度(TEM)=350.00,碰撞活化裂解(CAD)=Medium(以上单位均为仪器单位),离子选择通道分别为m/z 315.0/109.2(黄体酮)和m/z 299.0/109.1(内标)。结果:黄体酮血浆浓度在0.5～25 ng.mL-1范围内线性关系良好(r=0.9991),日内和日间精密度试验的RSD均小于7.5%,方法回收率为91.8%～100.7%,提取回收率为84.2%～91.5%,最低定量限为...     

注射用磷霉素钠中磷霉素钠二醇物含量测定方法的研究

目的:建立液相色谱串联质谱法和高效液相色谱-示差折光法测定注射用磷霉素钠中的磷霉素钠二醇物含量,并与现行中国药典方法比较,提高对药品的质量控制。方法:液相色谱串联质谱法采用Promosil CN(250 mm×4.6 mm,5μm)色谱柱,流动相为5 mmol.L-1醋酸铵-甲醇(82∶18),采用(-)ESI电离源,多反应监测(MRM)扫描方式,用于定量分析的离子分别为m/z 154.9→80.9(磷霉素二醇物)和m/z 120.9→77.1(内标苯甲酸)。高效液相色谱-示差折光法采用Agilent Zorb-ax NH2(250 mm×4.6 mm,5μm)色谱柱,流动相为10.89 g.L-1磷酸二氢钾溶液,流速为1.0 mL.min-1;柱温36℃;检测器温度为35℃。结果:采用液相色谱串联质谱法,二醇物在22.46～359.4 ng.mL-1范围内线性关系良好,定量限为4.492 ng.mL-1,每一样品的分析时间为5 min。采用高效液相色谱-示差折光法,二醇物在0.104～5.205 mg.mL-1浓度范围内线性关系良好,精密度为0.7%,重复性良好。结论:本研究2种方法专属性高、简便、可靠,可用于注射用磷霉素钠的质量研究和质量控制。     

壳聚糖树脂吸附提取辣椒素及其含量的测定

目的以干辣椒为原料,用壳聚糖树脂吸附提取辣椒碱并测定其含量。方法采用1%NaOH粗提取,提取液脱脂后用壳聚糖树脂柱吸附,乙醇洗脱、重结晶得淡黄色针状辣椒素的晶体。采用香兰素-亚硝酸钠比色法测定辣椒素含量,用HPLC分析晶体中辣椒碱和二氢辣椒碱的含量。结果辣椒素收率为0.38%,辣椒素晶体纯度为91.20%,其中辣椒碱为63.17%,二氢辣椒碱为26.75%。结论采用壳聚糖树脂吸附提取辣椒碱,方法简便,成本低廉,产率和纯度较高。     

HPLC法测定辣椒总碱中辣椒碱及二氢辣椒碱的含量

在 C6H5键合硅胶为填充剂的色谱柱上 ,以乙腈 - 0 .1% H3 PO4(33∶ 6 7)为流动相 ,检测波长为 2 80 nm,同时测定了辣椒总碱中辣椒碱及二氢辣椒碱的含量。结果表明 ,辣椒总碱中辣椒碱及二氢辣椒碱的含量约为5 8%和 30 % ,方法的平均加样回收率分别为 98.2 %和 97.6 %。     

Determination of capsaicin,nonivamide, and dihydrocapsaicin in blood and tissue by liquid chromatography-tandem mass spectrometry

A sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the analysis of capsaicin, nonivamide, and dihydrocapsaicin in blood and tissue has been developed. The method utilized a one-step liquid-liquid extraction that yielded an approximate 90% recovery of capsaicinoids from blood. Chomatographic separation of the capsaicinoids was achieved using a reversed-phase high-performance liquid chromatography column and a stepwise gradient of methanol and distilled water containing 0.1% (v/v) formic acid. Identification and quantitation of the capsaicinoids was achieved using electrospray ionization-tandem mass spectrometry monitoring the precursor-to-product-ion transitions for the internal standard octanoyl vanillamide (m/z 280 --> 137), capsaicin (m/z 306 --> 137), dihydrocapsaicin (m/z 308 -->137), and nonivamide (m/z 294 --> 137). Calibration curves, 1.0 to 250 ng/mL, were constructed by plotting concentration versus peak-area ratio (analyte/internal standard) and fitting the data with a weighted quadratic equation. The accuracy of the assay ranged from 90% to 107% for all analytes. The intra-assay precision (%RSD) for capsaicin was 4% at 2.5 ng/mL, 3% at 10 ng/mL, and 7% at 100 ng/mL. The interassay precision (% RSD) for capsaicin was 6% at 2.5 ng/mL, 6% at 10 ng/mL, and 7% at 100 ng/mL. Similar values for inter- and intra-assay precision were obtained for nonivamide and dihydrocapsaicin. This method was used to assay for capsaicinoids in blood and tissue samples collected from rats exposed to capsaicinoids via nose-only inhalation. The concentration of capsaicin in these samples ranged from < 1.0 to 90.4 ng/mL in the blood, < 5.0 to 167 pg/mg in the lung, and < 2.0 to 3.4 pg/mg in the liver.     

RP-HPLC法测定辣椒中辣椒碱的含量

目的:建立以反相高效液相色谱法测定辣椒中辣椒碱含量的方法。方法:色谱柱为Zorbax Eclipse XDB-C18(150mm×4.6mm,5μm),流动相为乙腈-0.1%磷酸溶液(45∶55,pH2.6),流速为1.0mL·min-1,柱温为40℃,检测波长为281nm。结果:辣椒碱检测浓度在30～70mg·L-1范围内与峰面积积分值呈良好线性关系(r=0.9999);平均加样回收率为101.33%,RSD=1.57%(n=9)。结论:本方法操作简便、快速,结果准确,可用于辣椒中辣椒碱的含量测定。     

Recent advances in the study on capsaicinoids and capsinoids

Chili peppers are the major source of nature capsaicinoids, which consist of capsaicin, dihydrocapsaicin, nordihydrocapsaicin, homodihydrocapsaicin, and homocapsaicin, etc. Capsaicinoids are found to exert multiple pharmacological and physiological effects including the activities of analgesia, anticancer, anti-inflammation, antioxidant and anti-obesity. Therefore, capsaicinoids may have the potential value in clinic for pain relief, cancer prevention and weight loss. In addition, capsaicinoids also display the benefits on cardiovascular and gastrointestinal system. It has been shown that capsaicinoids are potential agonists of capsaicin receptor or transient receptor potential vanilloid subfamily member 1 (TRPV1). They could exert the effects not only through the receptor-dependent pathway but also through the receptor-independent one. CH-19 Sweet peppers are the source of nature capsinoids, which share similar structure with capsaicinoids and consist of capsiate, dihydrocapsiate, and nordihydrocapsiate, etc, Comparing with capsaicinoids, capsinoids are less pungent and easily broken down in the normal aqueous conditions. So far, it has been found that capsinoids possess the biological properties of antitumor, antioxidant and anti-obesity. Since capsinoids are less toxic than capsaicinoids, therefore, capsinoids may have the advantages over capsaicinoids in clinical applications such as cancer prevention and weight loss.     

Effects of vehicle on the uptake and elimination kinetics of capsaicinoids in human skin in vivo

While the physiologic and molecular effects of capsaicinoids have been extensively studied in various model systems by a variety of administration routes, little is known about the uptake and elimination kinetic profiles in human skin following topical exposure. The present study evaluated the uptake and elimination kinetics of capsaicinoids in human stratum corneum following a single topical exposure to 3% solutions containing 55% capsaicin, 35% dihydrocapsaicin, and 10% other analogues prepared in three vehicles: mineral oil (MO), propylene glycol (PG), and isopropyl alcohol (IPA). Capsaicinoid solutions were evaluated simultaneously in a random application pattern on the volar forearms of 12 subjects using a small, single 150-microg dose. Capsaicin and dihydrocapsaicin were recovered from human skin using commercial adhesive discs to harvest stratum corneum from treated sites. Capsaicinoids were extracted from the stratum corneum-adhesive discs and quantified by liquid chromatography/mass spectroscopy (LC/MS). Both capsaicinoids were detected in stratum corneum 1 min after application with all vehicles and achieved a pseudo-steady state shortly thereafter. IPA delivered three times greater capsaicin and dihydrocapsaicin into the human stratum corneum than PG or MO at all time points investigated. The Cmax of capsaicin in IPA, PG, and MO was 16.1, 6.2, and 6.5 microg, respectively. The dihydrocapsaicin content was 60% of capsaicin with all vehicles. The estimated T(half) of capsaicin and dihydrocapsaicin in the three vehicles was similar (24 h). Thus, maximal cutaneous capsaicinoid concentrations were achieved quickly in the human stratum corneum and were concentration and vehicle dependent. In contrast, capsaicinoid half-life was long and vehicle independent.     

Effects of capsaicin and dihydrocapsaicin on human and rat liver microsomal CYP450 enzyme activities in vitro and in vivo

Capsaicin and dihydrocapsaicin, the two most abundant members of capsaicinoids in chili peppers, are widely used as food additives and for other purposes. In this study, we examined the inhibitory potentials of capsaicin and dihydrocapsaicin against CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4/5 activities in human liver microsomes. The effects of these two capsaicinoids on CYP450 enzymes were also evaluated in vivo in rats. The results demonstrated that capsaicin and dihydrocapsaicin moderately inhibited five isozymes (IC??) values ranging from 4.4 to 61.8 μM), with the exception of CYP2E1 (IC?? > 200 μM). Both capsaicinoids exhibited competitive, mixed, and noncompetitive inhibition on these isozymes (K (i) = 3.1 ± 0.5 - 78.6 ± 8.4 μM). Time-dependent inhibition of CYP3A4/5 by capsaicin was found. After multiple administrations of capsaicin and dihydrocapsaicin (1, 4, and 10 mg/kg) to rats, chlorzoxazone 6-hydroxylase activity and the expression of CYP2E1 were increased in liver microsomes. Our findings indicated that the possibility of food-drug interactions mediated by capsaicin and dihydrocapsaicin could not be excluded, and provided the useful information for evaluating the anticarcinogenic potentials of these two capsaicinoids.     

Effects of capsaicin and dihydrocapsaicin on human and rat liver microsomal CYP450 enzyme activities in vitro and in vivo

Capsaicin and dihydrocapsaicin, the two most abundant members of capsaicinoids in chili peppers, are widely used as food additives and for other purposes. In this study, we examined the inhibitory potentials of capsaicin and dihydrocapsaicin against CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4/5 activities in human liver microsomes. The effects of these two capsaicinoids on CYP450 enzymes were also evaluated in vivo in rats. The results demonstrated that capsaicin and dihydrocapsaicin moderately inhibited five isozymes (IC₅₀ values ranging from 4.4 to 61.8 μM), with the exception of CYP2E1 (IC₅₀>200 μM). Both capsaicinoids exhibited competitive, mixed, and noncompetitive inhibition on these isozymes (K _i = 3.1 ± 0.5 ? 78.6 ± 8.4 μM). Time-dependent inhibition of CYP3A4/5 by capsaicin was found. After multiple administrations of capsaicin and dihydrocapsaicin (1, 4, and 10 mg/kg) to rats, chlorzoxazone 6-hydroxylase activity and the expression of CYP2E1 were increased in liver microsomes. Our findings indicated that the possibility of food–drug interactions mediated by capsaicin and dihydrocapsaicin could not be excluded, and provided the useful information for evaluating the anticarcinogenic potentials of these two capsaicinoids.     

高效液相色谱荧光法测定家兔血浆中辣椒碱纳米乳的浓度及其药动学研究

目的:建立测定家兔血浆中辣椒碱纳米乳含量的方法,并考察其药动学参数。方法:采用双周期交叉试验设计,取6只家兔均分为2组,分别单剂量静脉注射辣椒碱纳米乳和辣椒碱溶液,剂量为1mg·kg-1,给药7d后交叉给药,分别于给药后3、6、9、12、15、25、40、60、90、120min时取血,以高效液相色谱荧光法测定其血药浓度,并计算药动学参数;色谱柱为Shimadzu VP-ODSC18,流动相为甲醇-水-磷酸(85∶15∶0.1),激发波长为280nm,检测波长为310nm。结果:辣椒碱检测浓度的线性范围为0.01974～4.934μg·mL-1(r=0.9995),最低检测限为4.9ng·mL-1,平均回收率为90.4%～113.8%(RSD≤10.1%);辣椒碱纳米乳和辣椒碱溶液的t1/2分别为(11.77±2.32)、(9.53±2.19)min,AUC0～∞分别为(5311±1324.6)、(4909.5±1110.4)ng·min·mL-1,cmax分别为315.7、300.8ng·mL-1,二者比较无显著性差异。结论:该法简便、准确、灵敏,适用于辣椒碱的血药浓度检测;辣椒碱制成纳米乳经静脉注射后基本上不影响其药动学参数。     

一测多评法测定天然辣椒碱中2种主要成分

目的建立以1种对照品同时测定天然辣椒碱中2种成分的质量评价方法。方法以辣椒辣素为对照,采用一测多评法,同时测定辣椒辣素和二氢辣椒辣素的含量。结果测得辣椒辣素与二氢辣椒辣素的相对校正因子为1.12,同时采用外标法和一测多评法测定样品中二氢辣椒辣素含量,采用t检验对二者测定值进行比较,二者之间无显著差异(P>0.05);以辣椒辣素的保留时间为1.00,计算得二氢辣椒辣素的相对保留时间为1.355,相对保留时间及保留时间差的RSD均小于5%。结论采用一测多评法,以辣椒辣素为对照,利用相对校正因子可实现同时测定辣椒辣素和二氢辣椒辣素的含量。