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同位素稀释液相色谱串联质谱法测定血清17α-羟孕酮候选参考测量程序的研究

中华检验医学杂志 2022年第5期45卷 449-455页

作者：虞科颖孙贺伟金中淦张素洁李卿居漪上海市临床检验中心参考测量实验室上海200126

目的研究1种基于液相色谱串联质谱(LC-MS/MS)技术的血清17α-羟孕酮(17α-OHP)候选参考测量程序并对其进行评价。方法2019年12月,取上海市临床检验中心员工体检留存的血清标本,采用正己烷乙酸乙酯混合液(体积比3∶2)提取加入同位素内标... 详细信息

目的研究1种基于液相色谱串联质谱(LC-MS/MS)技术的血清17α-羟孕酮(17α-OHP)候选参考测量程序并对其进行评价。方法2019年12月,取上海市临床检验中心员工体检留存的血清标本,采用正己烷乙酸乙酯混合液(体积比3∶2)提取加入同位素内标的血清样本,C18反向色谱柱分离,正离子电喷雾质谱仪检测,参照美国临床实验室标准化协会(CLSI)C62-A文件对建立的候选参考方法进行校准曲线、检测限与定量限、基质效应、不精密度、正确度、特异性、携带污染、线性范围等基本分析性能验证。结果17α-OHP的线性范围为0.21~119.67μg/L。检测限和定量限分别为5.218 ng/L、0.116μg/L。3种不同比例(50∶50、80∶20、20∶80)的血清与溶液混合物的相对基质效应分别为-0.02%、-0.40%、-0.90%。批内变异系数(CV)在0.164、14.81、81.63μg/L浓度处分别为1.73%~2.11%、0.98%~1.71%、0.47%~0.87%,批间CV在上述3个浓度点处分别为1.82%、1.03%、0.80%。加标样本平均回收率在0.5、20、100μg/L处分别为100.4%、101.7%、102.8%;检测中国计量科学研究院标准参考物质GBW09829,结果均在规定的不确定度范围内。无携带污染和特异性干扰。结论成功建立了基于同位素稀释LC-MS/MS技术的血清17α-OHP候选参考测量程序。该候选参考测量程序有着良好的精密度和准确度,能用于常规临床检验方法的量值溯源。

关键词： 17-α-羟孕酮质谱分析法放射性同位素稀释技术参考测量程序

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LAMB FETAL PULMONARY FLUID .1. VALIDATION AND SIGNIFICANCE OF METHOD FOR DETERMINATION OF VOLUME AND VOLUME CHANGE

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PEDIATRIC RESEARCH 1975年第4期9卷 190-195页

作者： SCARPELLI, EM CONDORELLI, S COSMI, EV ALBERT EINSTEIN COLL MED DEPT PEDIATPULM DIV1300 MORRIS PK AVEBRONXNY 10461 ALBERT EINSTEIN COLL MED DEPT ANESTHESIOLBRONXNY UNIV ROME ROMEITALY

An indicator-dilution method is described for measuring fetal pulmonary fluid, FPF, volume (Ve), and volume change with time (Vs) in the lamb fetus in utero. The indicator, albumin, which is the predominant protein of normal FPF, was administered directly to FPF in the form of radio-iodinated human serum albumin (RISA). We have shown that (1) RISA is distributed homogeneously throughout the FPF compartment, (2) it is not altered within FPF, (3) it does not alter the functional characteristics of FPF, and (4) it remains within the FPF compartment throughout the period of measurement. Ve varies directly with the weight of the fetus so that Ve per kilogram was 31.6-35.6 ml/kg in four of five fetuses. The Ve per kilogram was lower in one fetus at the start of the experiment but increased rapidly to 29.0 ml/kg within 45 min. Thus the relaxation volume of the fetus in utero is in the range of functional residual capacity (FRC) and thoracic gas volume of the air-breathing neonate, which indicates that the pulmonary transformation at birth is, in essence, an isovolumic change in state wherein the fetal liquid-lung becomes the neonatal air-lung. The rate at which FPF is formed (Vs) is about 1.5 ml/hr-kg; however, the rate may change in so far as both rapid increases and rapid decreases of FPF volume were observed. When breathing movements were induced in the fetus by stimulation of the sciatic nerve, FPF volume decreased rapidly (Vs equals -3.0 ml/hr-kg), which indicates that negative intrathoracic pressures promote its resorption. When FPF volume fell transiently or was low initially, there was subsequent rapid restoration of volume to the range of anticipated FRC.

关键词：血量体液电泳聚丙烯酰氨凝胶胎儿/分析肺/分析肺/胚胎学放射性同位素稀释技术坐骨神经血清白蛋白放射性碘标记绵羊硫喷妥钠/药理学子宫/解剖学和组织学动物女(雌)性妊娠

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LAMB FETAL PULMONARY FLUID .2. FATE OF PHOSPHATIDYLCHOLINE

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PEDIATRIC RESEARCH 1975年第4期9卷 195-201页

作者： SCARPELLI, EM CONDORELLI, S COLACICCO, G COSMI, EV ALBERT EINSTEIN COLL MED DEPT PEDIATPULM DIV1300 MORRIS PK AVEBRONXNY 10461 ALBERT EINSTEIN COLL MED DEPT ANESTHESIOLBRONXNY UNIV ROME ROMEITALY

Using the radio-iodinated human serum albumin ([131-I]-RISA) dilution method to measure lamb fetal pulmonary fluid (FPF) volume, we followed the disappearance of protein-complexed, 14-C-labeled phosphatidylcholine ([14-C]PC) during the first 90 min after its injection into FPF. The FPF samples were analyzed for total lipid 14-C activity and for distribution of 14-C in PC, other phospholipids (PL), fatty acids (FA), and neutral lipids (NL). For most sampling periods ascending aortic (AAo) and right atrial (RA) blood samples were obtained simultaneously with FPF and serum was analyzed for total lipid 14-C activity and for distribution of 14-C in total PL, FA, and NL. These studies indicate that (1) PC is cleared rapidly from FPF with an estimated half-time of 15-57 min; (2) FPF-PC may be metabolized to lyso-PC and FA within the fluid itself; and (3) FA derived from PFP-PC enter the pulmonary circulation, thus establishing a pulmonary arteriovenous FA gradient. The possible sites at which PC may be cleared from FPF are considered.

关键词：体液碳放射性同位素/诊断应用导管插入术上皮细胞脂肪酸类/分析胎儿/代谢脂类/分析肺/胚胎学肺/代谢磷脂酰胆碱类/代谢磷脂类/分析肺泡/细胞学放射性同位素稀释技术血清白蛋白放射性碘标记/诊断应用绵羊动物女(雌)性人类妊娠

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Quantitation of gluconeogenesis by ²H nuclear magnetic resonance analysis of plasma glucose following ingestion of ²H₂O

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ANALYTICAL BIOCHEMISTRY 2000年第1期277卷 121-126页

作者： Jones, JG Carvalho, RA Sherry, AD Malloy, CR Univ Texas SW Med Ctr Dept Radiol Mary Nell & Ralph B Rogers Magnet Resonance Ctr Dallas TX 75235 USA Univ Coimbra Dept Biochem Coimbra Portugal Univ Dallas Dept Chem Richardson TX USA Dept Vet Affairs Med Ctr Dallas TX USA

We present a simple H-2 MMR assay of the fractional contribution of gluconeogenesis to hepatic glucose output following ingestion of (H2O)-H-2. The assay is based on the measurement of relative deuterium enrichment in hydrogens 2 and 3 of plasma glucose. Plasma glucose was enzymatically converted to gluconate, which displays fully resolved deuterium 2 and 3 resonances in its H-2 NMR spectrum at 14.1 T. The signal intensity of deuterium 3 relative to deuterium 2 in the gluconate derivative as quantitated by H-2 NMR was Shown to provide a precise and accurate measurement of glucose enrichment in hydrogen 3 relative to hydrogen 2. This measurement was used to estimate the fractional contribution of gluconeogenesis to hepatic glucose output for two groups of rats;one group was fasted for 7 h and the other was fasted for 29 h, Rats were administered (H2O)-H-2 to enrich total body water to 5% over the last 4-5 h of each fasting period. For the 7-h fasted group, the hydrogen 3/hydrogen 2 enrichment ratio of plasma glucose was 0.32 +/- 0.09 (n = 7). This indicates that gluconeogenesis contributed 32 +/- 9% of total hepatic glucose output with glycogenolysis contributing the remainder, For the 29-h fasted group, the hydrogen 3/hydrogen 2 enrichment ratio of plasma glucose was 0.81 +/- 0.10 (n = 6), indicating that gluconeogenesis supplied the bulk of hepatic glucose output (81 +/- 10%). (C) 2000 Academic Press.

关键词：投药口服血糖/分析血糖/代谢氘氧化氘/投药和剂量氧化氘/药代动力学糖原异生磁共振波谱学/方法模型化学放射性同位素稀释技术大鼠 Sprague-Dawley 动物男(雄)性大鼠

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ABSORPTION, DISTRIBUTION, AND METABOLIC-FATE OF 7-CHLORO-3,3A-DIHYDRO-2-METHYL-2H,9H-ISOXAZOLO-(3,2-B)(1,3)-BENZOXAZIN-9-ONE IN RATS, DOGS, AND HUMANS

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JOURNAL OF PHARMACEUTICAL SCIENCES 1975年第8期64卷 1316-1321页

作者： EDELSON, J DOUGLAS, JF LUDWIG, BJ SCHUSTER, EB SHAHINIAN, S WALLACE LABS CRANBURYNJ 08512

The absorption and metabolic fate of 7-chloro-3,3a-dihydro-2-methyl-2H,9H-isoxazolo-(3,2-b)(1,3)-benzoxazin-9-one (I) was studied in rats, dogs, and humans. Orally administered I was readily absorbed by all species. In the rat, orally administered I was converted to its metabolite, 5-chlorosalicylic acid, by the intestinal wall. The half-lives of blood radioactivity, after the oral administration of I-9-14C, were about 18 and 12 hr in the rat and beagle hound, respectively. In human subjects, no intact I was detected in the bloodstream; however, the clearance of the metabolite, 5-chlorosalicylic acid, had a half-life of about 33 hr. Cleavage of the oxazine ring of I generated 5-chlorosalicylic acid, which was excreted both in the free form and conjugated with glycine and glucuronic acid. The isoxazole moiety was converted to beta-hydroxybutyric acid and its metabolites carbon dioxide and fumaric, citric, alpha-ketoglutaric, succinic, and malic acids. Binding of I to plasma proteins was extensive but was less than that of 5-chlorosalicylic acid.

关键词：消炎药/代谢药物稳定性肠吸收异恶唑类/代谢恶嗪类/代谢恶唑类/代谢蛋白质结合放射性同位素稀释技术水杨酸盐类/分离和提纯时间因素动物狗人类男(雄)性大鼠

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BODY-COMPOSITION IN MIDDLE-AGED WOMEN WITH SPECIAL REFERENCE TO THE CORRELATION BETWEEN BODY-FAT MASS AND ANTHROPOMETRIC DATA

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AMERICAN JOURNAL OF CLINICAL NUTRITION 1979年第7期32卷 1388-1395页

作者： NOPPA, H ANDERSSON, M BENGTSSON, C BRUCE, A ISAKSSON, B GOTHENBURG UNIV SAHLGRENS HOSP INST CLIN NUTR S-41345 GOTHENBURG SWEDEN

A variety of anthropometric measurements [used to determine obesity] was made in a randomized population sample of middle-aged women in 5 age strata in whom body composition was estimated from total body K and total body water determined by whole body counting and isotope dilution technique, respectively. No significant differences with age were found for total body K or total body water. A significant age difference was found for body fat mass. Simple linear correlations between anthropometric variables and body fat mass estimates were 0.90, 0.86, 0.77 for body weight, buttock circumference and sum of triceps and subscapular skinfold thicknesses, respectively. Multiple regression analysis showed that these 3 variables accounted for 80-91% of the variation in body fat in the different age strata studied. Multiple regression equations for prediction of body fat from anthropometric variables are given. In about 2/3 of the subjects, the difference between predicted body fat mass and estimated body fat was less than .+-. 2.5 kg.

关键词：脂肪组织/解剖学和组织学衰老人体测量术身体成分体质身高身体水份/分析体重钾/分析放射性同位素稀释技术皮肤壁厚度瑞典全身辐射量计数成年人老年人女(雌)性人类中年人

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A validated stable isotope dilution liquid chromatography tandem mass spectrometry assay for the trace analysis of cocaine and its major metabolites in plasma

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ANALYTICAL CHEMISTRY 1999年第10期71卷 2021-2027页

作者： Singh, G Arora, V Fenn, PT Mets, B Blair, IA Univ Penn Ctr Canc Pharmacol Dept Pharmacol Philadelphia PA 19104 USA

A validated method has been developed for the simultaneous quantitation of cocaine and its major metabolites (ecgonine methyl ester, benzoylecgonine, and norcocaine) in rat plasma. The method is based upon the use of stable isotope dilution liquid chromatography/atmospheric pressure chemical ionization/tandem mass spectrometry, Previously reported methods do not have the sensitivity and specificity that can be attained with this method. Plasma samples required no cleanup apart from protein precipitation, and no derivatization was required. Selected reaction monitoring was performed on the transitions of m/z 200 to m/z 182 (ecgonine methyl ester), m/z 290 to m/z 168 (benzoylecgonine), m/z 304 to m/z 182 (cocaine), and m/z 290 to m/z 168 (norcocaine), The standard curves were linear over the range from 2 ng/ml(benzoylecgonine, cocaine, and norcocaine) or 5 ng/mt (ecgonine methyl ester) to 1000 ng/mL in rat plasma. The lower limit of quantitation (LLQ) for benzoylecgonine, cocaine, and norcocaine was 2 ng/mL, and for ecgonine methyl ester, the LLQ was 5 ng/mL for plasma. This simple, rapid, reliable, and sensitive method of quantitation had excellent accuracy and precision for the four analytes, The method was sensitive enough to permit a detailed study of the pharmacokinetics of cocaine and its metabolites after administration of a bolus intravenous dose to rats.

关键词：生物转化色谱法高压液相可卡因/血液可卡因/药代动力学多巴胺摄取抑制剂/血液多巴胺摄取抑制剂/药代动力学质谱分析法放射性同位素稀释技术动物大鼠

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AN ANION-EXCHANGE RADIOASSAY FOR GLUCOSE-6-PHOSPHATE PHOSPHATASE - USE IN TOPOLOGICAL STUDIES WITH ENDOPLASMIC-RETICULUM VESICLES

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ANALYTICAL BIOCHEMISTRY 1992年第2期206卷 328-333页

作者： RUSH, JS WAECHTER, CJ UNIV KENTUCKY ALBERT B CHANDLER MED CTR COLL MED DEPT BIOCHEM LEXINGTON KY 40536 USA

A simple procedure is presented for the enzymatic preparation of [2-3H]mannose 6-phosphate (Man 6-P) with purified yeast hexokinase and unlabeled ATP. The enzymatically synthesized [2-3H]Man 6-P is utilized as the radiolabeled substrate in a new rapid assay for glucose 6-phosphate (Glc 6-P) phosphatase. The principle of the assay procedure is that the unreacted substrate, [2-3H]Man 6-P, is retained by the anion-exchange resin, AG 1-X8 (acetate), while the enzymatic product, [2-3H]-mannose, is eluted directly into a scintillation counting vial. When Glc 6-P phosphatase activity associated with mouse liver endoplasmic reticulum (ER) vesicles is assayed by the new chromatographic assay, the same characteristic latency and properties are observed, as determined by the commonly used colorimetric assay of inorganic phosphate produced. The anion-exchange radioassay described should be useful for a variety of topological studies on enzymes associated with membrane vesicles derived from liver and kidney ER.

关键词： B淋巴细胞/酶学脑/酶学 CHO细胞细胞培养的色谱法离子交换/方法仓鼠亚科内质网/酶学葡糖-6-磷酸酶/分析葡糖-6-磷酸酶/代谢己糖激酶同位素标记/方法动力学甘露糖磷酸盐类/生物合成甘露糖磷酸盐类/分离和提纯甘露糖磷酸盐类/代谢小鼠近交DBA 微粒体肝/酶学磷酸单酯水解酶类/分析磷酸单酯水解酶类/代谢放射性同位素稀释技术氚动物男(雄)性小鼠大鼠

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METHODOLOGICAL ASPECTS OF BLOOD-FLOW MEASUREMENT IN OVARIES CONTAINING CORPORA-LUTEA

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JOURNAL OF APPLIED PHYSIOLOGY 1975年第2期38卷 288-293页

作者： JANSON, PO ALBRECHT, I UNIV GOTHENBURG DEPT PHYSIOL GOTHENBURG SWEDEN UNIV GOTHENBURG DEPT OBSTET & GYNECOL GOTHENBURG SWEDEN CZECHOSLOVAK ACAD SCI INST PHYSIOL PRAGUE CZECHOSLOVAKIA

Ovarian blood flow measurements were carried out in pseudopregnant anesthetized rabbits in order to compare three different techniques. Direct measurement by venous cannulation and flow determination by 15-mum radioactive microspheres resulted in values 5 and 10 times higher, respectively, than indicator fraction technique using -86Rb. The direct method inevitably involves surgical handling of the ovary and its vessels, which may account for the lower values found with this method than with microspheres. A comparison between the dynamics of -86Rb uptake in the luteal ovary and the hindleg tissues, mainly consisting of striated muscle, indicated that this technique will underestimate ovarian blood flow due to a low ovarian extraction of the radionuclide. In the preparation for direct measurement the relation between blood pressure and flow was linear. This lack of autorgeulation is an important factor to consider in studies of ovarian steroid secretion rates. The radioactive microsphere technique appears to reflect physiological blood flow in the ovary and corpus leteum of the rabbit and can be useful in studies of vascular mechanisms in this gland.

关键词：血流速度/方法血压导管插入术黄体/生理学评价研究(主题) 微球体肌/血液供给卵巢/血液供给假孕放射性同位素稀释技术局部血流铷动物女(雌)性兔

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CARDIOPULMONARY BLOOD VOLUMES AT REST AND DURING MUSCULAR EXERCISE MEASURED BY M-113 IN RADIOCARDIOGRAPHY

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ACTA PHYSIOLOGICA SCANDINAVICA 1975年第2期95卷 145-152页

作者： KUIKKA, J PYORALA, T LEHTOVIRTA, P REKONEN, A CENT HOSP JYVASKYLA DEPT RADIOTHERAPYJYVASKYLAFINLAND

Cardiac output, stroke volume, right heart and left heart blood volume, pulmonary blood volume and pulmonary mean time were measured at rest and during muscular exercise in 14 healthy subjects in the supine position. The method used was 113mIn radiocardiography measured precordially with a single detector. The results were analyzed with a minicomputer by a modified gamma function fitting method. The increase in cardiac output during exercise was 140 per cent (p is less than 0.001). The stroke volume increased 15 per cent (p is less than 0.05) in subjects with heart rates less than 160 beats/min, and decreased in subjects with heart rates greater than or equal to 160 beats/min. The right heart blood volume decreased from 115 ml/m2 to 105 ml/m2 to 215 ml/m2 (5 per cent; p is less than 0.1) and the left heart blood volume decreased from 120 ml/m2 to 110 ml/m2 (5 per cent; p is less than 0.01). The total heart blood volume thus decreased from 235 ml/m2 (5 per cent; p is less than 0.05). The pulmonary blood volume showed a corresponding increase from 250 ml/m2 to 280 ml/m2 (12 per cent; p is less than 0.005), and the cardiopulmonary blood volume did not markedly change (485 ml/m2 and 495 ml/m2). Pulmonary mean time decreased from 5.0 s to 2.4 s (52 per cent; p is less than 0.001).

关键词：血液循环时间血量心排血量用力心脏功能试验心率肺循环放射性同位素稀释技术成年人女(雌)性人类男(雄)性

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