比色法测定辣椒总碱凝胶剂中主药含量

目的　采用比色法测定辣椒总碱凝胶剂中辣椒总碱的含量。方法　将样品经NaOH(pH 12 5 )碱化后 ,加 0 9% 4 氨基安替比林溶液和 5 4 %铁氰化钾溶液 ,摇匀 ,在 2 5℃± 2℃的暗处放置 180min ,测定吸收度 ,测定波长为 4 70nm。结果　辣椒素在 70 32～ 2 11 0 μg·ml 1浓度范围内 ,r=0 9996 ,回收率为 98 94 % ,RSD为 1 4 %。结论　本法操作简便准确 ,可作为测定辣椒总碱制剂中辣椒总碱含量的方法。     

RP-HPLC同时测定代温灸膏中肉桂酸、桂皮醛、辣椒碱、二氢辣椒碱及6-姜酚的含量

目的:建立用高效液相色谱法同时测定代温灸膏中主要药效成分肉桂酸、桂皮醛、辣椒碱、二氢辣椒碱及6-姜酚的含量。方法:采用Alltima C18(4.6 mm×250 mm,5μm)色谱柱,流动相为乙腈-0.1%磷酸,梯度洗脱,流速1 mL·min-1,检测波长280nm,柱温30℃。结果:肉桂酸、桂皮醛、辣椒碱、二氢辣椒碱、6-姜酚浓度分别在5.66～42.45,20.40～153.0,16.20～121.5,10.16～76.20,13.12～98.40μg.mL范围内与峰面积呈良好的线性关系。上述化合物的回收率和RSD分别为98.2%,1.4%;100.0%,1.0%;99.2%,1.1%;99.2%,1.2%;100.1%,1.0%。结论:该方法能准确、快速分析代温灸膏中5个主要成分,灵敏度高,重现性好,可用于控制代温灸膏的质量。     

定氮法测定鱼藤酮辣椒碱悬浮剂中辣椒碱含量

目的:建立鱼藤酮-辣椒碱悬浮剂辣椒碱含量的化学定氮分析方法.方法:定氮法测定悬浮剂中辣椒碱含量.结果用空白对照及蛋白质氮含量进行校正.结果:定氮法测定辣椒碱含量均值为0.25 g/100 mL(精密度RSD 1.77%),回收率100.67%(RSD 1.65%),蛋白质对辣椒碱含量测定的影响在误差控制范围内.结论:本方法操作简便,重现性好,可作为鱼藤酮-辣椒碱悬浮剂中辣椒碱质量控制方法.     

RP-HPLC法同时测定辣椒碱透皮贴剂中4种生物碱类成分的含量

目的建立RP-HPLC法同时测定辣椒碱透皮贴剂中辣椒碱、二氢辣椒碱、降二氢辣椒碱和高二氢辣椒碱4种生物碱类成分含量。方法采用Kromasil C_(18)(250 mm×4.60 mm,5μm)柱,以乙腈-水(体积比为42∶58)为流动相,流速为1.0 mL·min~(-1),柱温为40℃,检测波长为280 nm。结果 4种生物碱类成分的色谱峰与邻近色谱峰均能良好的分离,辣椒碱和二氢辣椒碱的质量浓度分别在2.65～79.50 mg·L~(-1)(r=0.999 7)、1.08～64.80 mg·L~(-1)(r=0.999 9)内与峰面积呈良好的线性关系,平均回收率分别为100.2%(RSD=1.5%)、99.5%(RSD=0.7%)。结论该方法可用于辣椒碱透皮贴剂的质量控制。     

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。     

比色法测定辣椒总碱凝胶剂中主药含量

目的 采用比色法测定辣椒总碱凝胶剂中辣椒总碱的含量。方法 将样品经NaOH(pH12.5)碱化后,加0.9%4-氨基安替比林溶液和5.4%铁氰化钾溶液,摇匀,在25℃±2℃的暗处放置180min,测定吸收度,测定波长为470nm。结果 辣椒素在70.32～211.0μg·ml^-1浓度范围内,r=0.9996,回收率为98.94%,RSD为1.4%。结论 本法操作简便准确,可作为测定辣椒总碱制剂中辣椒总碱含量的方法。     

反相高效液相色谱法测定辣椒风湿软膏中总辣椒碱含量

目的用反相高效液相色谱法（RP—HPLC法）测定辣椒风湿软膏中总辣椒碱的含量。方法采用ZorbaxC18柱（250mm×4．6mm，5pm）；流动相为甲醇-2．5％乙酸溶液（60：40），流速为2．0mL／min，检测波长为280nm，进样量为20μL。结果方法的线性范围为2-200μg／mL（r=1．0000），精密度RSD为0．45％（n=5），加样回收率为98．64％（RSD为0．72％）。结论RP～HPLC法具有专属、灵敏、快速、准确等优点，可用于辣椒风湿软膏中总辣椒碱含量的测定。     

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。     

HPLC测定辣椒风湿贴中辣椒碱的含量

目的 建立测定辣椒风湿贴中辣椒碱含量的方法.方法 采用HPLC法,Luna C18色谱柱(250 mm×4.6 mm,10 μm),甲醇-水(65:35)为流动相,检测波长281 nm.结果 在6.04～241.60 μg·ml-1范围内,辣椒碱浓度与峰面积的线性关系良好(r=0.9997),RSD=0.95%(n=6),平均回收率为100.6%(n=5).结论 所建方法准确、简便、快速,适用于辣椒风湿贴的质量控制.     

辣椒碱膜剂镇痛作用的研究

目的：评价辣椒碱膜剂（LM）的镇痛作用及皮肤刺激性。方法：以无水乙醇为溶剂，大豆磷脂、维生素E和胆酸钠为成膜材料制备LM；采用热板法和福尔马林法评价LM的镇痛作用；用家兔皮肤评价Ⅲ的皮肤刺激性。结果：按12．5mg/kg、25mg/kg剂量分别给药．LM的镇痛作用显著高于辣椒碱乙醇溶液，P〈0．01；LM对皮肤的刺激性显著低于辣椒碱乙醇溶液。结论：LM对热刺激和炎症引起的疼痛具有显著的镇痛作用；大豆磷脂和胆酸钠能显著提高辣椒碱的镇痛作用，同时显著减轻辣椒碱对皮肤的刺激性。     

Gastrointestinal absorption and metabolism of capsaicin and dihydrocapsaicin in rats

Gastrointestinal absorption of capsaicin and dihydrocapsaicin was studied in rats in vivo and in situ. Rapid absorption of capsaicin or dihydrocapsaicin from stomach and small intestine occurred in vivo. About 85% of the dose was absorbed in the gastrointestinal tract within 3 hr. In situ, within 60 min after the administration of capsaicin and dihydrocapsaicin into stomach, jejunum, and ileum, about 50, 80, and 70% of the respective dose had disappeared from the lumen. When 2,4-dinitrophenol or NaCN was added, no significant reduction in uptake of [3H]dihydrocapsaicin was observed in the jejunum. These results suggested that capsaicin and its analogs were absorbed by a nonactive process in jejunum. [3H]Dihydrocapsaicin was mainly absorbed via the portal system but not a mesenteric lymphangial one. The radioactivity in the portal blood was composed of 85% of [3H]dihydrocapsaicin and 15% of its metabolite (8-methyl nonanoic acid) bound to the albumin fraction. Dihydrocapsaicin-hydrolyzing enzyme activity was found in jejunal tissue. These results suggest that capsaicin and its analogs partly received a first-pass effect, i.e., metabolism of a compound following first absorption in the gastrointestinal tract. It is concluded that capsaicin and its analogs are readily transported to the portal vein through the gastrointestinal tract by a nonactive process and partly metabolized during absorption.     

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.     

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。     

Dihydrocapsaicin-induced hypothermia and substance P depletion

Administration of dihydrocapsaicin to rats resulted in a dose-dependent (0.5-10 mg/kg s.c.) hypothermia. Dihydrocapsaicin was approximately 65% more effective in producing hypothermia than capsaicin. Desensitization and cross-tolerance occurred to the hypothermic effects of both capsaicin and dihydrocapsaicin. Repeated administration of either dihydrocapsaicin or capsaicin resulted in chemogenic antinociception but not marked thermal antinociception. In addition, repeated administration of dihydrocapsaicin, like capsaicin, resulted in depletion of substance P from dorsal root ganglia and dorsal spinal cord but not from the hypothalamus, corpus striatum or ventral spinal cord. These data indicate that dihydrocapsaicin, or radiolabelled dihydrocapsaicin, may be a useful tool for investigating the mechanisms by which capsaicin alters thermoregulation and primary afferent neuron function.     

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.     

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.     

微波辅助萃取-高效液相色谱-串联质谱法测定辣椒中3种辣椒素

目的:建立微波辅助萃取-高效液相色谱-串联质谱法测定辣椒中3种辣椒素类化合物(辣椒素、二氢辣椒素、降二氢辣椒素)含量。方法:辣椒样品经微波辅助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%甲酸水溶液和乙腈,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:3种辣椒素的检出限为0.01～0.11 ng.mL-1,定量限为0.03～0.36 ng.mL-1,辣椒素、二氢辣椒素、降二氢辣椒素平均回收率分别为95.4%,94.3%,93.1%;RSD分别为2.1%,2.5%,2.7%。结论:该方法简便、准确、灵敏,可用于辣椒中3种辣椒素的分析。     

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。