高效液相色谱-蒸发光散射测定三七中三七皂苷R1及人参皂苷Rg1、Re、Rb1的含量

目的:用 HPLC—ELSD 测定三七药材中三七皂苷 R_1及人参皂苷 Rg_1、Re、Rb_1的含量。方法:色谱柱为 Lichrospher NH_2柱(4.6 mm×250 mm,5μm),流动相为乙腈-水(80:20);漂移管温度为90℃,载气流速为2.1 L·min~(-1)。结果:三七皂苷 R_1及人参皂苷 Rg_1、Re、Rb_1分别在0.316～1.58μg、1.21～6.05μg、0.448～2.24μg及1.49～7.47μg呈良好线性关系;药材中4种成分的平均回收率(n=6)分别为102.2%(RSD=2.6%)、99.4%(RSD=2.2%)、101.8%(RSD=2.5%)及97.6%(RSD=2.4%)。结论:该方法简便、准确,分离效果好,无干扰,可用于三七药材的质量评价。     

RP—HPLC梯度洗脱法测定血塞通片中三七皂苷R1、人参皂苷Rb1及人参皂苷Rg1的含量

目的:用高效液相色谱梯度洗脱法同时检测血塞通片中三七皂苷R_1、人参皂苷Rb_1和人参皂苷Rg_1 3种皂苷的含量。方法:用Eclipse XDB—C_8分析柱;乙腈-水二元梯度洗脱;0—15 min(25:75—32:68),平衡5 min;流速1.0 mL·min~(-1);检测波长203 nm。结果:三七皂苷R1、人参皂苷Rg_1和人参皂苷Rb_1的线性范围分别为O.51-3.05,1.68—10.09,2.73—16.39μg。该方法回收率三七皂苷R_1为99.92%(RSD=0.70%)、人参皂苷Rg_1为98.93%(RSD=1.4%)、人参皂苷Rb_1为102.8%(RSD=0.81%)。结论:HPLC梯度洗脱法能将多种皂苷很好地分离检测,提高了时效,减少了误差,结果表明该方法准确可靠,重现性好,结果稳定。     

高效液相色谱法测定三七-丹参药对不同配比水煎液中3种成分溶出率的变化

目的采用HPLC法研究三七-丹参药对不同配比水煎液中三七皂苷R_1、人参皂苷Rg_1和人参皂苷Rb_1 3种成分溶出率的变化规律。方法采用ZORBAX SB-C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈-水,梯度洗脱;流速1 mL·min~(-1);检测波长203 nm;柱温35℃。结果三七-丹参药对配伍比例为2∶1时,3种成分的总溶出率最高,而配伍比例为1∶3时溶出率最低。药对配伍比例为1∶2时,丹参对三七皂苷R_1、人参皂苷Rg_1的溶出率影响最大,而3∶1时对溶出率影响最小。配伍比例为2∶1时,丹参对人参皂苷Rb_1溶出率影响最大,而1∶3时对溶出率影响最小。结果显示,丹参、三七合煎可影响三七有效成分的溶出,可能两者合煎产生了新的化合物。结论三七-丹参不同的配伍比例会影响3种成分的溶出,以2∶1配比时最利于3种成分的溶出,2∶1是最佳配伍比例。     

脑得生片中10种成分的定量研究

目的建立HPLC法,对脑得生片中3′-羟基葛根素、葛根素、葛根素芹菜糖苷、3′-甲氧基葛根素、大豆苷、染料木苷、三七皂苷Rb_1、人参皂苷Rg_1、大豆苷元和人参皂苷Rb_1 10种有效成分进行定量分析。方法色谱条件:色谱柱为Kromasil C_(18)柱(250 mm×4.6 mm,5μm);流动相为乙腈-2.0 g·L~(-1)磷酸水溶液,梯度洗脱;流速为0.8 mL·min~(-1);检测波长为250和203 nm;柱温为35℃。结果 10个成分的峰具有良好的分离度;3′-羟基葛根素、葛根素、葛根素芹菜糖苷、3′-甲氧基葛根素、大豆苷、染料木苷、三七皂苷Rb_1、人参皂苷Rg_1、大豆苷元和人参皂苷Rb_1的线性范围分别为14.99～299.88,15.00～300.03,8.26～165.13,10.24～204.82,10.01～200.13,6.40～128.00,15.75～314.93,21.00～419.97,7.01～140.14和17.50～349.97μg·mL~(-1),相关系数分别为0.999 5,0.999 8,0.999 9,0.999 5,0.999 6,0.999 5,0.999 6,0.999 8,0.999 6和0.999 6;精密度、稳定性与重复性实验的RSD值均小于1.60%;10个对照品的加样回收率均在97.10%～98.48%之间,RSD值均在0.76%～1.56%之间;10批制剂中3′-羟基葛根素、葛根素、葛根素芹菜糖苷、3′-甲氧基葛根素、大豆苷、染料木苷、三七皂苷Rb_1、人参皂苷Rg_1、大豆苷元和人参皂苷Rb_1的平均含量分别为7.43,13.33,8.29,10.64,9.35,1.04,2.21,8.44,1.73和2.97 mg·g~(-1),RSD值分别为1.23%,0.82%,1.11%,1.24%,1.07%,1.58%,1.28%,0.86%,1.45%和1.28%。结论脑得生片中10种主要有效成分的含量检测方法操作简便、准确、可靠、稳定且重复性好,为该制剂的质量控制提供了有效的评价方法。     

HPLC法测定人参、西洋参和三七不同部位中人参皂苷的含量

目的:为了探讨人参、西洋参和三七中人参皂苷的资源含量,以确保人参、西洋参和三七中人参皂苷资源充分被利用,为开发以人参皂苷为主的创新药物提供科学数据。方法:采用高效液相色谱法对人参、西洋参和三七不同部位中人参皂苷的含量进行测定。色谱条件为:Agilent 1100 Series 高效液相色谱仪;色谱柱为德国 Nucleosil-C_(18)(4.6 mm×150 mm,5μm);流动相为水-乙腈梯度洗脱,流速1.5 mL·min~(-1);A为水,B为乙腈;梯度洗脱程序为:0～16 min,56%B;16～20 min,56%→100%B;20～38 min,100%B;38～45 min,100%→56%B;45～60 min,56%B;60～70 min,56%B。所有组分均70 min 内出完。检测波长203 nm,柱温35℃,灵敏度为0.02AUFS。线性关系考察r=0.9994;精密度试验 RSD=0.26%,平均回收率为99.88%,重现性试验 RSD=2.0%,分离度为R=3.042。结果:人参须根含有较高的人参皂苷 Rb_1(1.082%),人参茎叶则含有较高的人参皂苷 Rc(1.002%)、Re(3.430%)和 Rg_1(1.303%);西洋参根含有较高的人参皂苷 Rb_1(2.213%)和 Re(0.9188%),西洋参芦头中含有较高的人参皂苷 Rb_1(2.840%)和Re(1.224%);三七根含有较高的人参皂苷 Rb_1(2.163%)和 Rg_1(2.633%),三七芦头中含有较高的人参皂苷 Rb_1(4.376%)和 Rg_1(4.145%)。结论:高效液相色谱法分离、分析人参皂苷效果好、准确、迅速、简便,也可作为评价人参属植物质量的有效分析方法。建议对人参、西洋参和三七中含量较高的人参皂苷进行提取分离,直接用于创新药物的开发。     

RP-HPLC法同时测定西洋参含片中4种成分

目的建立同时测定西洋参含片中人参皂苷Rg_1、人参皂苷Re、人参皂苷Rb_1及拟人参皂苷F11含量的RP-HPLC方法。方法色谱柱为Agilent TC-C18(250 mm×4.6 mm,5μm);流动相为0.5 mL·L~(-1)磷酸溶液(A)-乙腈(B),梯度洗脱;流速:1.0mL·min~(-1);检测波长:203nm;柱温:30℃。结果人参皂苷Rg_1和人参皂苷Re及人参皂苷Rb_1在2.5~500μg·mL~(-1)、拟人参皂苷F11在5.0~1 000μg·mL~(-1)范围内线性关系良好(r≥0.999 4);平均加样回收率均大于95.0%。结论该方法简便、准确,重复性好,可作为西洋参含片的质量控制方法。     

RP-HPLC法同时测定舒胸颗粒中3种皂苷类成分的含量

目的建立RP-HPLC法同时测定舒胸颗粒中三七皂苷R_1、人参皂苷Rg_1、人参皂苷Rb_1的含量测定方法。方法采用色谱柱Diamosil C_(18)(4.6×250mm,5μm),柱温30℃,流动相乙腈-水,梯度洗脱,检测波长203nm。结果 3个皂苷类成分色谱峰面积与对照品进样量均呈良好的线性关系,r≥0.9993;样品加样回收率在98.9%~99.7%之间,RSD均小于1.4%。结论该方法灵敏度高、专属性好、操作简便、重复性好,可为舒胸颗粒质量控制提供依据。     

胶束电动毛细管色谱法分离分析三七中的人参皂苷Rg1、人参皂苷Re和三七皂苷R1

目的:用胶束电动毛细管色谱法分离测定三七中人参皂苷 Rg_1、人参皂苷 Re 和三七皂苷 R_1 3种皂苷含量。方法:采用Beckman MDQ 电泳仪,40 cm×50 μm的熔融石英毛细管,有效柱长30 cm,缓冲液为20 mmol·L~(-1)SDS、40 mmol·L~(-1)胆酸钠、60 mmol·L~(-1)硼砂,检测波长200 nm。结果:人参皂苷 Rg_1低、中、高剂量的平均加样回收率分别为101.5%,100.6%,100.4%,RSD 分别为2.4%,2.2%,1.0%;人参皂苷 Re 低、中、高剂量的平均加样回收率分别为101.3%,99.23%,99.66%,RSD 分别为2.0%,1.3%,1.1%;三七皂苷 R_1低、中、高剂量的平均加样回收率分别为100.6%,100.5%,100.7%,RSD 分别为1.1%,1.1%,0.86%。结论:本方法能够将三七药材中人参皂苷 Rg_1、人参皂苷 Re 和三七皂苷 R_1完全分离,具有灵敏度高、快速、易于重现且所用试剂价格低廉的特点,因此适合于中药三七的测定,并对今后三七的鉴别和质量控制具有重要意义。     

炮制前处理对三七中三七皂苷含量的影响

目的:研究前处理对三七主根中皂苷含量的影响,为研究三七炮制工艺及探讨生熟三七功效异同提供物质基础。方法:采用蒸制法将三七主根炮制成熟三七,高效液相色谱法测定10种单体皂苷含量。Vision HT C₁₈柱(4.6 mm×250 mm,5μm),乙腈和水梯度洗脱,流速1.0 ml·min^-1,柱温24℃,检测波长203 nm。结果:三七经蒸制后皂苷成分三七皂苷R₁、人参皂苷Rg₁、Re、Rb₁、Rd含量均有不同程度的降低,而人参皂苷Rh₁、Rh₄、Rk₃、20(S)-Rg₃、20(R)-Rg₃含量均有不同程度的增加;炮制品中5种皂苷成分含量下降程度和新产生成分含量增加程度与蒸制时间、温度相关;皂苷成分在120℃条件下降解和形成的速度比105℃快,总体上降解和形成的快慢:蒸制鲜三七主根切片>蒸制干三七主根整体>蒸制鲜三七主根整体,其中,Rg₁降解最快,Rh₄形成最快。结论:前处理方法影响皂苷成分降解和形成的快慢,高温高压对皂苷成分有一定的破坏作用,该方法可用于三七炮制品中皂苷类成分的含量测定和质量控制,为三七"生打熟补"与物质变化之间的相关性研究提供依据。     

HPLC法测定红药胶囊中三七皂苷R1及人参皂苷Rg1的含量

目的:建立红药胶囊中三七皂苷 R_1及人参皂苷 Rg_1的含量测定方法。方法:采用 Alltima—C_(18)(4.6 mm×150 mm,5μm),流动相为乙腈-0.1%磷酸溶液(20:80),流述为1.0 mL·min~(-1),检测波长为203 nm;柱温为40℃,外标法测定。结果:三七皂苷 R_1和人参皂苷 Rg_1进样量分别在0.345～2.76μg(r=0.9999)和0.945～7.56μg(r=0.9999)范围内与峰面积呈良好线性关系;平均回收率(n=6)分别为98.6%和99.2%。结论:本法简便可靠,专属性强,可用于红药胶囊的质量检测。     

The aminomethylation of 1-cyano-isochromane and 1-cyano-isothiochromane

The Aminomethylation of 1-Cyano-isochromane and 1-Cyano-isothiochromane Aminomethylation of 1-cyano-isochromane (1a) and 1-cyano-isothiochromane (1i) can be achieved via reaction of carbanions 2 with α-haloamines 3 . Dialkylaminomethyl and dialkylaminobenzyl compounds 4 are formed.     

Preferential induction of CYP1A1 and CYP1B1 in CCSP-positive cells.

Both benzo[a]pyrene (BaP) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are potent ligands of aryl hydrocarbon receptors (AhR). Although animal studies indicate that both compounds induce pathological changes in the peripheral lung, the specific cell type involved remains unclear. Clara cells, expressing Clara cell specific protein (CCSP) and abundant in cytochrome P450, are nonciliated bronchiolar epithelial cells in the peripheral lung. Here we explore the hypothesis that CCSP-positive Clara cells are highly responsive to AhR ligands and are the primary cell type involved in BaP- and TCDD-induced toxicities. The responsiveness to AhR ligands was evaluated by measuring the respective mRNA and protein levels of cytochrome P450 1A1 (CYP1A1) and 1B1 (CYP1B1) using real-time RT-PCR and immunocytochemistry assays. Two in vitro models were used: primary cultures of human small airway epithelial (SAE) cells and rat lung slice cultures. In the presence of calcium, human SAE cells differentiated into CCSP-positive cells. BaP- and TCDD-induced mRNA and protein levels of CYP1A1 and CYP1B1 levels were significantly elevated in CCSP-positive cell cultures. Similarly, AhR mRNA and protein levels were increased in CCSP-positive cell cultures, as determined by real-time RT-PCR and Western blot analysis. When rat lung slice cultures were treated with BaP or TCDD for 24 h, CYP1A1 and CYP1B1 proteins were strongly induced in Clara cells. These results indicate that, in the peripheral lung of both rats and humans, CCSP-positive cells (Clara cells) may be more sensitive to AhR ligands than other cell types.     

Influence of genetic polymorphisms of CYP1A1 and GSTM1 on the urinary levels of 1-hydroxypyrene

The aim of the present study is to evaluate the influence of the genetic polymorphism of two enzymes involved in the biotransformation of xenobiotics, cytochrome P450 1A1 (CYP1A1) and glutathione-S-transferase M1 (GSTM1), on the urinary levels of 1-hydroxypyrene (1-OH-P) in workers exposed to polycyclic aromatic hydrocarbons (PAHs) and in unexposed workers (controls). The study group consisted of 30 controls recruited among employees of a service company and 171 PAHs-exposed workers from two electric steel plants and an iron foundry (all males, ranging between 18 and 60 years of age). Determination of airborne PAHs and urinary 1-OH-P was performed by high-performance liquid chromatography (HPLC) with fluorimetric detection. Polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) was used to determine the genetic polymorphisms of CYP1A1 (CYP1A1*2A and CYP1A1*2B) and GSTM1. No influence of the genetic polymorphism of CYP1A1 and GSTM1 on the urinary levels of 1-OH-P was observed in this study.     

Toxicokinetics and Bioavailability of Oral and Intravenous 1, 1 -Dichloroethylene

Toxicokinetics and Bioavailability of Oral and Intravenous 1,1-Dichloroethylene.PUTCHA, L., BRUCKNER, J. V., D'SOUZA, R., DESAI, F., AND FELDMAN,S. (1986). Fundam. Appl. Toxicol. 6, 240–250. Althoughaliphatic halocarbons have been identified as contaminants ofdrinking water supplies, little definitive information is availableon their gastrointestinal (G.1) absorption and toxicokinetics.Therefore, a study of a representative halocarbon, 1,1-dichloroethylene(1,1-DCE), was undertaken to contrast the kinetics of the chemicalfollowing iv injection with that following oral administration.Four dosage-levels of 1,1-DCE (10, 25, 50, and 100 mg/kg BW)in 50% aqueous polyethylene glycol 400 were given iv and poto fasted and nonfasted male Sprague—Dawley rats. Serialblood samples were taken from the tail artery of the lightlyetherized animals for up to 490 min after dosing. 1,1-DCE concentrationsin the whole blood were determined by gas chromatographic head-spaceanalysis. Evaluation of the iv data revealed that disappearanceof 1,1-DCE from the systemic circulation followed a triexponentialpattern. Light ether anesthesia did not appear to alter thepharmacokinetics of iv-injected 1,1-DCE. There was no differencebetween nonfasted and fasted iv rats in biological half-life(t) or in any other pharmacokinetic parameter. Total body clearance,t apparent volume of distribution and volume of distributionin the central compartment did show increases with increasingdose in these animals. Oral dosing experiments revealed that1,1-DCE was absorbed very rapidly and completely from the G.I.tract. Peak blood levels were reached 2 to 8 min following oraladministration of 1,1-DCE as an aqueous suspension. The t of1,1-DCE in orally dosed rats was somewhat longer than in theiriv counterparts The t values for nonfasted, orally dosed ratswere longer than for their fasted counterparts, suggesting delayedabsorption due to the presence of food in the G.I. tract. Bioavailabilityof 1,1-DCE, as determined by comparing areas under blood concentrationversus time curves (AUCs), was equivalent in animals given thesame dose of 1,1-DCE iv and po. AUCs increased with increasingdose in iv and po groups, but the increases were not proportionalto dose.