高效液相色谱法同时测定赤丹丸中6种有效成分的含量

 目的建立高效液相色谱法测定赤丹丸中丹参素钠、黄芩苷、黄芩素、汉黄芩素、汉黄芩苷和丹参酮ⅡA含量的方法。方法Agilent Zorbax Extend-C₁₈(4.6mm×250mm,5μm)柱;流动相组成为A:甲醇,B:水(磷酸调pH3.0),C:乙腈,进行梯度洗脱;流速1.0mL·min^-1;检测波长280nm。结果丹参素钠线性范围16.4～327.0mg·L^-1,平均回收率为97.5%,RSD为1.31%;黄芩苷线性范围15.8～316.0mg·L^-1,平均回收率为97.4%,RSD为1.02%;黄芩素线性范围10.3～206.0mg·L^-1,平均回收率为99.3%,RSD为0.68%;汉黄芩素线性范围12.6～251.0mg·L^-1,平均回收率为100.2%,RSD为1.76%;汉黄芩苷线性范围11.3～225.0mg·L^-1,平均回收率为99.4%,RSD为2.6%;丹参酮ⅡA线性范围5.04～100.7mg·L^-1,平均回收率为97.8%,RSD为1.4%;结论本方法测定结果准确可靠,可用于测定赤丹丸中6成分的含量。     

折光法快速判断中药柱层析过程中的始终点及其在树脂精制丹参有效部位中的应用

目的:介绍运用折光原理快速判断中药柱层析过程中始终点的方法及其在大孔树脂精制丹参有效部位的应用。方法:通过丹参提取物经AB-8型大孔吸附树脂精制有效部位,水-30%乙醇-80%乙醇梯度洗脱,间隔取样,在线监测洗脱液折光率并分析变化规律,同时以高效液相色谱法和紫外可见分光光度法分别检测丹酚酸B、丹参萜类成分含量作参比,判断洗脱过程的始点和终点。结果:丹参有效部位精制过程中洗脱的始终点为:水洗终点第4BV(折光率0),即30%乙醇洗脱始点,30%乙醇洗脱终点第8BV(折光率10.5),即80%乙醇洗脱始点,80%乙醇洗脱终点第10.75BV(折光率19.5)。结论:折光法准确可靠,便捷快速,在中药柱层析处理过程有很强的实践意义。     

PREDICTION OF THE SEVERITY OF ABO HAEMOLYTIC DISEASE OF THE NEWBORN BY CORD BLOOD TESTS

ABSTRACT. Whyte, J. and Graham, H. (Bellshill Maternity Hospital, Bellshill, Lanarkshire, Scotland). Prediction of the severity of ABO haemolytic disease of the newborn by cord blood tests. Acta Paediatr Scand, 70:217, 1981. –Seventy-one ABO incompatible (heterospecific) infants and 71 controls, who were free from other potential causes of jaundice, were studied to ascertain which cord blood tests reliably predict the severity of ABO haemolytic disease of the newborn (ABO HDN). The modified Direct Antiglobulin Test (spin DAGT) was positive in all infants who required treatment for haemolytic jaundice and only DAGT positive children showed evidence of impending haemolytic anaemia or compensated haemolysis in cord or capillary blood. Cord serum bilirubin concentration had some predictive value, particularly when the level exceeded 85 µmol/l, but it was a less reliable indicator and had greater value if used in association with the DAGT. The elution test, which is frequently used as a diagnostic tool in ABO HDN, had no predictive value and we felt that its putative value is due to overdiagnosis of ABO HDN in jaundiced heterospecific infants. We conclude that the spin DAGT, despite the weakness of the reaction, reliably identifies infants at risk from severe ABO HDN and is sufficiently sensitive to be used as a single screening test for the early detection of the disorder.     

FBL-3肿瘤抗原的研究:Ⅱ.从多肽池中分析肿瘤抗原

在抗肿瘤免疫效应中细胞毒T淋巴细胞(CTL)是通过识别肿瘤细胞MHC I类分子结合的肿瘤源性多肽.为分析肿瘤细胞的抗原性及肿瘤抗原的多肽性质,本文选用FBL-3肿瘤细胞,用pH3.3的枸橼酸-磷酸盐缓冲液间隔24～36小时多次酸洗肿瘤细胞,以获得MHC I类分子结合的多肽,并分析酸洗液中多肽混合物的肿瘤抗原性,以及RP-HPLC分离的部分收集液多肽的肿瘤抗原性,为FBL-3肿瘤细胞抗原的确定提供基本数据.结果表明:酸洗法能有效获得肿瘤细胞MHC I类分子相关的多肽抗原,肿瘤细胞多肽抗原的分离、纯化、HPLC分析以及抗原性鉴定是肿瘤抗原性确定的一个重要方法.     

离子色谱法测定浓维磷糖浆中甘油磷酸根及磷酸根的含量

摘 要 目的：建立抑制电导离子色谱法同时测定浓维磷糖浆中有效成分甘油磷酸根及磷酸根的含量。方法: 采用AG19 HC（50 mm×4 mm）保护住和IonPac AS18 HC（250 mm×4 mm）阴离子分析柱,以氢氧化钾溶液梯度淋洗,检测器为电导检测器,检测方式为抑制电导检测。结果:在该色谱条件下,甘油磷酸根峰与磷酸根峰均能有效分离,线性范围分别为1.054～84.35 μg·ml-1（r=0.999 4）及0.316 8～25.35 μg·ml-1（r=0.999 8）;平均回收率分别为99.35%（RSD=1.16%,n=9）和98.96%（RSD=2.61%,n=9）。结论:本法简便,准确,专属性强,有助于更好地控制本品的质量。     

双波长梯度洗脱高效液相色谱法测定冲和软膏中芍药苷和蛇床子素的含量

目的:建立RP-HPLC法同时测定冲和软膏中芍药苷和蛇床子素含量的方法。方法:色谱柱为Phenomisl C18柱(150 mm×4.6 mm5,μm);以乙腈-水为流动相进行梯度洗脱;流速为1.0 mL·min-1;柱温为35℃;检测波长为230和330 nm。结果:芍药苷与蛇床子素的线性范围分别为3.12～249.8μg·mL-1(r=0.999 8)和1.54～123μg·mL-1(r=0.999 9);平均加样回收率分别为101.0%(RSD=1.07%,n=9)和100.8%(RSD=1.21%,n=9)。结论:本方法操作简便,测定结果准确,重复性好,可作为该产品质量控制的方法。     

梯度洗脱HPLC法测定苄星青霉素中的有关物质

目的建立测定苄星青霉素有关物质的梯度洗脱HPLC法。方法采用端基封尾的C18柱,流动相A为0.05mol/L磷酸二氢钾溶液(用磷酸调节pH值至3.1),流动相B为甲醇,流速1.0mL/min,线性梯度洗脱,检测波长220nm,柱温40℃。结果二苄基乙二胺和青霉素分别与相邻杂质峰分离完全,二苄基乙二胺的检测限为2.5ng,青霉素的检测限为0.6ng。结论本法专属、灵敏,可作为苄星青霉素有关物质的测定方法。     

HPLC分离西沙必利有关物质的方法学研究

 目的建立西沙必利有关物质分离的HPLC方法。方法采用二元泵,在室温下以流速1.0 mL·min-1进行线性梯度洗脱,以(A)乙腈(B)0.05 mol·L-1磷酸盐缓冲液(pH7.5)组成流动相,检测波长为275 nm,进样量10μL或20μL。结果与氟哌啶醇分离度＞3,西沙必利、有关物质Ⅰ和有关物质Ⅱ线性良好(r=0.999)。最小检出量有关物质Ⅰ为0.025μg·mL-1,有关物质Ⅱ为0.05μg·mL-1,日内日间差异分别为6.9%和6.6%。结论本法灵敏、专属,可有效分离有关物质及降解产物,可用于有关物质及降解产物检查。     

Immunoaffinity isolation of native membrane glucocorticoid receptor from S-49++ lymphoma cells

The membrane glucocorticoid receptor (mGR), previously correlated with glucocorticoid-induced lymphocytolytic competency, was purified under nondenaturing conditions from mGR-enriched mouse S-49 T lymphoma cells. Proteins were immunoaffinity batch adsorbed to BUGR-2 monoclonal antibody-coupled protein A Sepharose 4B beads, and elution by epitope competition was compared with standard denaturation procedures. Elution with BUGR-2 epitope peptides released multiple mGRs (42–150 kDa) and heat shock proteins 70 and 90, suggesting that mGR interacts with these protein chaperones under physiological conditions. The mGR-heat shock protein 90 interaction was inhibited by 1 μM geldanamycin. Several other mGR binding partners were captured and most were dissociated from mGR by 0.6M salt. Peptide maps of purified mGR displayed immunoreactive bands unique to mGR. Scatchard analysis estimated a k_d value of 239 nM and a B_max of 384 fmol/mg protein for mGR, compared to a k_d of 19.5 nM and a B_max of 90.3 fmol/mg protein for the intracellular GR (iGR). The rank order of affinities for mGR were RU-486 > dexamethasone > triamcinolone acetonide=aldosterone. Other steroids had no significant binding affinity. These results show that epitope-purified mGR on the plasma membrane of mouse lymphoma cells is similar but not identical to iGR.     

高效液相色谱梯度洗脱法测定大鼠胆汁中9-硝基喜树碱含量

目的建立大鼠胆汁中 9 硝基喜树碱的高效液相色谱梯度洗脱方法。方法胆汁样品经乙腈 水 ( 1∶1 )稀释后 ,直接进样。检测波长为 370nm ,采用HypersilBDSC18柱 ( 5 μm,1 5 0mm× 4 6mmI D )分离 ,使用以乙腈 2 %甲酸水溶液为流动相的二元梯度洗脱方法。结果线性范围为 0 5～2 5 μg/mL,定量限为 0 5 μg/mL。结论本法操作简便 ,成功应用于胆汁中 9 硝基喜树碱的含量的测定及胆汁中 9 硝基喜树碱代谢物的初步研究。