粪便内中短链脂肪酸柱前衍生化UHPLC-QTOF-MS/MS分析方法研究

建立柱前衍生化超高效液相色谱-串联四级杆飞行时间质谱(UHPLC-QTOF-MS/MS)定性、定量表征小鼠粪便内中短链脂肪酸的方法,并用于评价其在抗生素处置前后小鼠粪便样品中的变化。动物实验方案获得江苏省中医药研究院动物实验伦理委员会的批准。以3-硝基苯肼为衍生化试剂,对衍生化反应条件、色谱及质谱条件进行优化,建立了粪便中16个中短链脂肪酸的定性、定量方法。所有分析物在各自浓度范围内线性关系良好(R2>0.99),日内、日间精密度RSD均小于10%,重复性RSD小于6%,加样回收率介于80%~120%之间,样品36 h内稳定性RSD小于7%;联合抗生素处置后小鼠粪便内的中短链脂肪酸种类和含量均发生了明显改变。其中,甲酸、乙酸、丙酸、丁酸、异丁酸、戊酸、乳酸等含量明显降低,而庚酸和丁二酸的含量明显增加。该方法准确、可靠,可作为粪便内中短链脂肪酸的表征方法。     

高效液相色谱法同时测定牙痛药水中丁香酚和胡椒碱含量

目的建立同时测定牙痛药水中丁香酚和胡椒碱含量的高效液相色谱(HPLC)法。方法色谱柱为C18柱(150 mm×4.6 mm,5μm),流动相为甲醇-水(72∶28),检测波长为280 nm,流速为1.0 m L/min,柱温为30℃。结果2种待测成分分离度良好,阴性无干扰。丁香酚、胡椒碱质量浓度线性范围分别为25.4~279.8μg/m L(r=0.9996)和12.5~137.8μg/m L(r=1.0000);平均回收率分别为100.53%和98.99%,RSD分别为0.82%和0.67%(n=9);丁香酚、胡椒碱含量范围分别为0.850~1.439 mg/m L和0.227~0.632 mg/m L。结论该方法操作简便、快速,结果准确,重复性好,可同时测定牙痛药水中丁香酚和胡椒碱的含量。     

HPLC同时测定丙戊酸钠口服溶液中丙戊酸钠及防腐剂的含量

目的 建立高效液相色谱(HPLC)同时测定丙戊酸钠口服溶液中丙戊酸钠及防腐剂含量的方法。方法 采用十八烷基键合硅胶色谱柱(Thermo Scientific BDS Hypersil C18,5 μm,250 mm×4.6 mm),以0.01mol/l磷酸二氢钠(磷酸调节pH为2.3)-乙腈(63：37)为流动相,等度洗脱,流速1.0 mL/min,检测波长210 nm、254nm。结果 丙戊酸钠线性范围为0.99 ~2.92 mg/ml,r=0.99991;回收率为98.86 %~100.30 %,总体RSD=0.5 %(n=9);羟苯甲酯钠线性范围为14.74μg/ml～58.96μg/ml,r=0.99994;回收率为100.59 %~101.57 %,总体RSD=0.4 %(n=15);羟苯丙酯钠线性范围为10.21μg/ml～20.41μg/ml,r=0.99995;回收率为99.78%~101.85 %,总体RSD=0.8 %(n=12)。结论 本方法简单、迅速、可靠,可用于丙戊酸钠口服溶液的检测和质量控制。     

HPLC法检测减肥类保健食品中非法添加的酚酞和盐酸西布曲明

目的:建立高效液相色谱法检测减肥类保健食品中非法添加的酚酞和盐酸西布曲明。方法:采用高效液相色谱法,色谱柱为Agilent TC-C18色谱柱(150 mm×4.6 mm,5μm),流动相为0.02 mol/L乙酸铵溶液(用乙酸调节p H至4.0)-甲醇(45∶55),流速为1.0 ml/min,柱温为35℃,检测波长为225 nm。结果:酚酞和盐酸西布曲明分别在10.77~107.70μg/ml(r=0.999 9)和11.15~111.50μg/ml(r=0.999 9)范围内,浓度与其峰面积呈良好线性关系,平均回收率分别为99.37%(RSD为0.73%)和98.97%(RSD为0.60%)。结论:本方法简便易行、准确可靠,可用于检测减肥类保健食品中非法添加的酚酞和盐酸西布曲明。     

UPLC-MS/MS法同时检测抗风湿中药制剂中非法添加的12种非甾体抗炎药

目的:建立同时检测抗风湿中药制剂中非法添加的12种非甾体抗炎药。方法:采用超高效液相色谱-质谱法。色谱条件:色谱柱为Hypersil Golden C18,流动相为5 mmol/L甲酸铵溶液-甲醇(梯度洗脱),流速为0.2 m L/min,柱温为40℃,进样量为2μL。质谱条件:离子源为电喷雾离子源,气帘气压为25 k Pa,雾化气压为60 k Pa,辅助气压为55 k Pa,电喷雾电压为4 500 V,离子源温度为650℃,采集方式为多反应监测模式。结果:对乙酰氨基酚、乙酰水杨酸、氨基比林、美洛昔康、布洛芬、萘普生、舒林酸、尼美舒利、双氯芬酸、吲哚美辛、酮洛芬、塞来昔布检测质量浓度线性范围分别为0.01~2.0μg/m L(r=0.995 6)、0.05~5.0μg/m L(r=0.997 6)、0.01~2.0μg/m L(r=0.998 7)、0.02~5.0μg/m L(r=0.995 0)、0.02~5.0μg/m L(r=0.995 3)、0.02~5.0μg/m L(r=0.996 5)、0.05~5.0μg/m L(r=0.995 4)、0.02~5.0μg/m L(r=0.996 0)、0.05~5.0μg/m L(r=0.995 9)、0.02~5.0μg/m L(r=0.9957)、0.02~5.0μg/m L(r=0.996 8)、0.01~2.0μg/m L(r=0.998 7);定量限≤0.20 mg/g,检测限≤0.05 mg/g;精密度、稳定性、重复性试验的RSD<5.0%;加样回收率为80.8%~114.2%(RSD为3.85%~7.32%,n=9)。结论:该方法操作简便,精密度、稳定性、重复性好,可用于抗风湿中药制剂中非法添加的12种非甾体抗炎药的同时检测。     

HPLC梯度洗脱法同时测定地麻滴鼻液中2种组分的含量

目的建立测定地麻滴鼻液中地塞米松磷酸钠和盐酸麻黄碱的高效液相色谱法。方法色谱柱为Agilent ZORBAX SB-C18柱(150 mm×4.6 mm,5μm),流动相为磷酸二氢钾缓冲液-甲醇-乙腈,梯度洗脱,流速1 m L/min,检测波长242 nm。结果地塞米松磷酸钠在5.2~52.0μg/m L范围内线性关系良好(r=0.999 9),盐酸麻黄碱在0.501~5.01 mg/m L范围内线性关系良好(r=0.999 5)。地塞米松磷酸钠平均回收率为99.13%,RSD为1.46%;盐酸麻黄碱平均回收率为99.19%,RSD为1.04%。结论该法简便、准确并且重现性好,可用于地麻滴鼻液中地塞米松磷酸钠和盐酸麻黄碱的质量控制。     

高效液相色谱法测定盐酸氟西汀胶囊含量

目的:建立反相高效液相色谱法测定盐酸氟西汀胶囊的含量.方法:采用kromasil C18色谱柱(5μm,4.5mm×15mm)为分离柱;流动相为甲醇:0.05mol/L磷酸二氢钾缓冲液(55:45);流速:1mL/min;检测波长:230nm;柱温:室温.结果:盐酸氟西汀在52.5mg/L～315mg/L浓度范围内线性良好(r=0.9994);方法平均回收率为99.66%(RSD=0.98%,n=5);5次测定的精密度0.16%～0.35%.结论:本方法操作简便、快速、准确、灵敏,适用于盐酸氟西汀胶囊的含量测定.     

超高效液相色谱串联质谱法同时测定走川骨刺酊中3种乌头类生物碱含量

目的建立同时测定走川骨刺酊中3种乌头类生物碱含量的超高效液相色谱-串联质谱(UPLC-MS/MS)法。方法色谱柱为ACQUITY UPLCBEH C18柱(50 mm×2.1 mm,1.7μm),流动相为0.1%甲酸溶液-乙腈(梯度洗脱),流速为0.25 m L/min,柱温为35℃,进样量为10μL;采取电喷雾离子源(ESI),正离子模式,采集方式为多反应监测(MRM)。结果乌头碱、次乌头碱、新乌头碱质量浓度分别在0.9198~91.9800 ng/m L(r=0.9978,n=7),1.048~104.800 ng/m L(r=0.9974,n=7),1.268~126.800 ng/m L(r=0.9998,n=7)范围内与峰面积线性关系良好,加样回收率分别为104.22%,105.05%,104.34%,RSD分别为3.13%,1.63%,1.62%(n=6)。结论该方法操作简便,精密度、重复性好,专属性高,结果准确,可用于同时测定走川骨刺酊中3种乌头类生物碱的含量。     

HPLC法同时测定氨咖黄敏胶囊中二组分的含量

目的建立高效液相色谱法同时测定氨咖黄敏胶囊中对乙酰氨基酚、咖啡因2种成分的含量。方法采用Agilent extend-C_(18)(150 mm×4.6 mm,5μm)色谱柱,流动相为乙腈(A)-三乙胺溶液(1 m L三乙胺加79 m L水,调p H至3.1,B)(17∶83),流速1.0 m L/min,检测波长217 nm;柱温25℃,进样量15μL。结果对乙酰氨基酚、咖啡因的线性浓度范围分别为831.33~2 078.33μg/m L(r=0.999 6)和50.27~125.67μg/m L(r=0.999 4),两组分平均回收率分别为99.99%(RSD=0.02%)和100.14%(RSD=0.52%)。结论该方法灵敏度、准确度高,重复性好,适用于氨咖黄敏胶囊中对乙酰氨基酚和咖啡因2种有效成分的测定。     

鹅不食草中短叶老鹤草的定量方法研究

目的:建立鹅不食草中有效成分短叶老鹤草的定量分析方法. 方法:采用反相高效液相色谱法测定含量,色谱柱为Kromasil C18色谱柱(250mm×4.6mm,5μm),流动相为乙腈-水(45:55),流速:1.0mL/min;柱温:30℃;进样量:10μL;检测波长:225nm.结果:短叶老鹤草在0.010065~0.30195mg/mL范围内线性关系良好(r=1.000), 平均回收率(n=6) 为99.53%, 样品含量范围为0.048%~0.43%. 结论:建立的方法简单灵敏,准确可靠,重复性好,可用于鹅不食草的质量控制.     

Comparison of the choleretic properties of bile acids

The choleretic properties of cholic, chenodeoxycholic, and deoxycholic acid and their taurine and glycine conjugates were compared to their ability to form micelles. It has previously been concluded that deoxycholate has the lowest critical micellar concentration; chenodeoxycholate is slightly higher and cholic is much higher. Conjugation with glycine and taurine has little or no effect on the critical micelle concentration. Since the choleretic properties of bile salts are thought to be directly proportional to their osmotic activities, one might suspect that deoxycholic acid would be the least choleretic, chenodeoxycholic slightly more choleretic and cholic much more choleretic, with little difference between the conjugated and unconjugated forms. However, in the present study, cholic, chenodeoxycholic and taurocholic acid produced similar increases in bile flow (450–700 μl/kg) after an equimolar dose (55 μM/kg). Except for the conjugation of deoxycholic acid with taurine, conjugation of these bile acids with glycine or taurine always decreased the choleretic properties of the bile acids. Therefore, it has been concluded that there is not a good correlation between the in vitro osmotic properties of bile acids and their ability to increase bile flow.     

抗癌活性成分满五酸等三个化合物的分离和鉴定

从产于四川宜宾翼梗五味子果实中,分得抗癌活性成分满五酸(1),同时分得schizandronic acid(2)和cuparenic acid(3),结构由与标准化合物比较光谱数据和物理常数而确证。     

HPLC法测定熊去氧胆酸片有关物质的含量

[摘要]目的：建立高效液相色谱法测定熊去氧胆酸片有关物质含量的方法。方法：用十八烷基硅烷键合硅胶为填充剂，以磷酸盐缓冲液（pH3.0）-甲醇-乙腈（35:37:28）为流动相，示差折光检测器测定。结果：在选定的色谱条件下，各成分可达到很好的分离，7-酮基胆石酸、胆酸、鹅去氧胆酸、胆石酸分别在0.05~2.00，0.05~2.00，0.05~2.00，0.10~2.00mg·mL-1的范围内线性关系良好(r>0.9989)；精密度、稳定性试验的RSD均不大于0.5%；回收率分别为100.1%、99.8%、100.7%和101.9%（n=9,RSD<1.4%）。7-酮基胆石酸、胆酸、鹅去氧胆酸、胆石酸定量限分别为3.0、2.5、7.0、20μg·mL-1，检测限分别为0.8、0.7、2.0、3.5μg·mL-1。测定供试品两批，供试品一7-酮基胆石酸检出量为0.03%，胆酸和胆石酸均未检出，鹅去氧胆酸检出量为0.66%；供试品二7-酮基胆石酸、鹅去氧胆酸检出量均为0.10%，胆酸和胆石酸均未检出。结论：该方法灵敏度高，专属性强，适于测定熊去氧胆酸片中有关物质的含量。     

Pomegranate (<Emphasis Type="Italic">Punica granatum</Emphasis>) pure chemicals show possible synergistic inhibition of human PC-3 prostate cancer cell invasion across Matrigel™

Summary Four pure chemicals, ellagic acid (E), caffeic acid (C), luteolin (L) and punicic acid (P), all important components of the aqueous compartments or oily compartment of pomegranate fruit (Punica granatum), and each belonging to different representative chemical classes and showing known anticancer activities, were tested as potential inhibitors of in vitro invasion of human PC-3 prostate cancer cells in an assay employing Matrigel™ artificial membranes. All compounds significantly inhibited invasion when employed individually. When C, P, and L were equally combined at the same gross dosage (4 μ g/ml) as when the compounds were tested individually, a supradditive inhibition of invasion was observed, measured by the Kruskal-Wallis non-parametric test.An erratum to this article can be found at     

HPLC同时测定升麻中三个酚酸类成分的含量

目的建立快速测定升麻中咖啡酸、阿魏酸及异阿魏酸含量的HPLC方法。方法色谱柱为C18柱(4.6 mm×250 mm,5μm);流动相:乙腈-0.01%磷酸梯度洗脱;柱温:30℃;检测波长:320 nm;流速:1.0 ml/min。结果咖啡酸、阿魏酸、异阿魏酸分别在5.78~115.6μg/ml(r=0.9998,n=6),4.03~80.60μg/ml(r=1,n=6),16.45~329.00μg/ml(r=1,n=6)范围内线性关系良好,回收率在97.6%~101.5%。结论建立的方法准确可靠,可以用于测定升麻中咖啡酸、阿魏酸、异阿魏酸的含量。     

Antitumor triterpenes from medicinal plants

Thirteen kinds of naturally occurring or derivatised triterpenes, reported to have an antitumoral property, were reinvestigated on the basis of their direct cytotoxicity or the inhibitory activity on cell growth against five kinds of cultured human tumor cells,i. e., A-549, SK-OV-3, SK-MEL-2, XF498 and HCT15,in vitro. Ursonic acidIII, betulinic acidVIII, betulonic acidX and glycyrrhetinic acidXI were exhibited a marked inhibition on cell growth.     

Oral antipyretic therapy: Evaluation of the N-aryl-anthranilic acid derivatives mefenamic acid,tolfenamic acid and flufenamic acid

Summary The antipyretic activity of three N-aryl-anthranilic acid derivatives, mefenamic acid, tolfenamic acid and flufenamic acid, was compared and their optimal antipyretic dose determined in a trial in 87 children (aged 5 months to 15 years), who suffered from infections and fever exceeding 38.5°C. Tolfenamic acid proved to be the most potent antipyretic agent of the three drugs; it was eight times more powerful than mefenamic acid and three times more powerful than flufenamic acid. The optimal antipyretic doses were: mefenamic acid 4 mg/kg, tolfenamic acid 0.5 mg/kg and flufenamic acid 1.5 mg/kg. It is evident that the antipyretic activity of these anthranilic acid derivatives is even greater than their antirheumatic effect, the difference being most noticeable in the case of tolfenamic acid.     

HPLC测定L-天门冬氨酸中的有关物质

目的建立L-天门冬氨酸中羟基丁二酸、丁二酸和富马酸等有关物质的检查方法。方法采用HPLC法,使用AQ C18色谱柱,0.01 mol.L-1的磷酸二氢钾水溶液(磷酸调pH3)为流动相,检测波长200 nm。结果主成分和杂质可完全分离。结论建立的方法灵敏、专属,可用于L-天门冬氨酸中有关物质的测定。