Automated immunochemical method for determination of urinary protein of plasma origin.

An automated continuous-flow procedure has been developed for the rapid determination of urinary proteins of plasma origin. Antiserum to whole human plasma was used as the reagent, and the antigen-antibody reactions were quantitated by nephelometry. By adding polyethylene glycol (mol wt 6000-7500) to the reaction medium, reaction time was decreased to less than 3 min; no sample blanks were required; and samples were analyzed at a rate of 70 per hour. Recovery studies yielded an average of 98.5% of the added protein. In-run replicate precision (CV) of the method was 1.45%; day-to-day precision was 2.58%.     

液相色谱-串联质谱法检测人血浆中碘帕醇

目的建立一种稳定的检测人血浆碘帕醇的液相色谱-串联质谱法(LC-MS/MS),并对该方法进行性能评价。方法以氘代同位素作为内标,人血浆样品经直接沉淀法前处理后进样,用XSelect^TM HSS PFP色谱柱进行分离,用含0.1%甲酸-5mmol/L九氟戊酸-10%乙腈的水溶液等度洗脱;正离子电喷雾离子化扫描检测,多反应监测(MRM)扫描分析,建立检测血浆碘帕醇的LC-MS/MS方法。根据CLSI EP10-A和CLSI C62-A,评价该方法的线性范围、正确度、精密度、选择性、基质效应、稳定性。结果 LC-MS/MS检测血浆碘帕醇的线性范围为1～1 000μgI/mL(以含碘量计算);低、中、高浓度(3、50、800μgI/mL)血浆质控品检测结果与理论靶值的偏移为-6.1%～7.5%,重复性以不精密度表示,CV<6.6%(4.6%～6.6%),期间精密度CV<8.5%(6.6%～8.5%),方法回收率为92.5%～95.9%,基质效应为88.8%～95.2%;方法选择性良好,溶血和脂血因素均不影响检测结果。血浆样品在室温下放置最好不要超过48 h,在-80℃下反复冻融不超过2次,经前处理后的血浆样品4℃自动进样器中放置不超过72 h,碘帕醇可稳定检测。结论建立并评价了一种可靠的检测人血浆碘帕醇的LC-MS/MS方法。     

Automated method for the determination of angiotensin-converting enzyme in serum

A method for the determination of angiotensin-converting enzyme in serum (S-ACE; EC 3.4.15.1.) with use of 3-(2-furylacryloyl)-L-phenylalanyl-glycyl-glycine (FAPGG) as substrate has been adapted for the Cobas Bio microcentrifugal analyser. The method allows 24 determinations per hour in a sample volume of 28 microliters with a within run precision of less than 3% and a between run precision of less than 5%. The reaction is linear up to at least 470 U/l. The reference interval in 92 blood donors has been determined to 14-110 U/l. The method correlates well with the manual method of Hurst & Lovell-Smith (r = 0.982). We have found the method excellently suited for routine assay.     

Automated method for the determination of angiotensin-converting enzyme in serum

A method for the determination of angiotensin-converting enzyme in serum (S-ACE; EC 3.4.15.1.) with use of 3-(2-furylacryloyl)-L-phenylalanyl-glycyl-glycine (FAPGG) as substrate has been adapted for the Cobas Bio microcentri-fugal analyser. The method allows 24 determinations per hour in a sample volume of 28 μl with a within run precision of less than 3% and a between run precision of less than 5%. The reaction is linear up to at least 470 U/l. The reference interval in 92 blood donors has been determined to 14–110 U/l. The method correlates well with the manual method of Hurst & Lovell-Smith [4] (r=0.982). We have found the method excellently suited for routine assay.     

Automated cyanide-free method for haemoglobin determination on Technicon H.1

We have developed a cyanide-free haemoglobin method for our Technicon H.1 counter. By adding 2.5 mmol/l of the ionic surfactant sodium dodecyl sulphate in 154 mmol/l sodium chloride, the haemoglobins are converted to a stable product, denatured globin haemichrome, within the 25 s available on the instrument. The method is as precise and accurate as the original H.1 method, and the reagent is easily made in the laboratory at a very low price. Since the method is relatively safe with regard to waste disposal, the effluent from the counter can be drained into the laboratory sink. The method, a 'SDS-haemichrome method', has now worked very satisfactorily in routine use for a year. Manufacturers of haematological analysers should consider this haemoglobin method for their automated instruments.     

Automated method for the direct determination of total serum cholesterol

An automated method for determining total cholesterol in serum is described which does not require a preliminary solvent extraction. A special glass unit has been constructed and details of the preparation of cholesterol acetate standards in acetic acid are given. Thirty samples, including five standards, can be estimated per hour and the volume of sample required is 0.5 ml.     

Automated determination of fluvoxamine in plasma by column-switching high-performance liquid chromatography.

A column-switching system with high-performance liquid-chromatographic separation and ultraviolet detection is described for automated determination of fluvoxamine in human plasma or serum. Samples were injected and the drug was retained in a clean-up column [20 x 4.6 mm (i.d.)] filled with C8 reversed-phase material (10-micron particles). After unwanted material was washed out, the drug was eluted and separated with an analytical chromatography column, 4.6 x 250 mm (i.d.), filled with Nucleosil 100 CN (5-micron particles) with an acetonitrile:methanol:0.01 mol/L phosphate buffer eluent (188:578:235 by vol) at a flow rate of 1.5 mL/min for < 20 min and detected by spectrometry at 214 nm. With oxaprotiline as internal standard, fluvoxamine could be easily quantified, and it was well separated from endogenous plasma constituents and various psychoactive drugs. The detection limit was 10 micrograms/L (31.6 nmol/L), the analytical recoveries were 97-100%, and the relationship between drug concentration and detector response was linear from 0 to 1000 micrograms/L (3160 nmol/L). The automated method is suitable for therapeutic monitoring of fluvoxamine in the treatment of psychiatric patients.     

High-dose methadone and the need for drug measurements in plasma.

We report a case of high-dose methadone prescribed to a heroin addict for pain control. The patient was prescribed methadone during convalescence from surgery and subsequently for maintenance treatment. Dosing was started at 360 mg of methadone per day and reduced over 12 days to an 80 mg/day maintenance dose. Although the patient was drowsy on the initial dose, his recovery was uneventful. However, there were complaints of pain and withdrawal discomfort when the plasma concentration decreased to less than 1 mg/L. Measurements of methadone in plasma were helpful for monitoring the recovery of this patient after surgery and are likely to prove useful in similar cases.     

Automated determination of total plasma cholesterol: a serum calibration technique.

The automated (AutoAnalyzer II) determination of cholesterol in nine serum pools in the concentration range 3.465-8.871 mmol/l, gave results that were approximately 10% higher than reference values when the analyses were based on unesterified cholesterol standards containing the same amount of water as the sample extracts (1963 analyses in 12 laboratories during a 12 month period; automated value = 0.032 + 1.10 X (Reference value)). A serum calibration procedure was successful ilues, and was equally effective in correcting the values observed for aliquots of 368 fresh-frozen plasma samples analyzed in each of the 12 laboratories during a 38 month period.