Quantitation of phenylalanine and tyrosine from dried Blood/Plasma spots with impregnated stable isotope internal standards (SIIS) by FIA-SRM.

BACKGROUND: Dried Blood Spot (DBS) analysis has been used for identification and quantification of diseases and disorders in large populations. Simply collecting blood or plasma samples on cotton paper, followed with an organic solvent extraction, many small molecules can be detected and quantified. In a typical procedure of DBS analysis in newborn screening, stable isotope internal standards (SIIS) are added to extraction solvent as a reference. However, this way of employing SIIS does not reflect extraction efficiency, or protein binding issues, nor does it reflect potential degradation that could occur. In addition, punched-out discs from larger DBS are known to have imprecision typically ≥ 15%. METHODS: We developed and tested an approach, internal quantitative DBS (iqDBS), which delivers an exact volume of whole blood or plasma to a paper disc that is impregnated with a dried concentration of SIIS for quantitation. Amino acids were derivatized to make butyl esters and measured using Flow Injection Analysis with Selected Reaction Monitoring (FIA-SRM). RESULTS: We demonstrated with phenylalanine and tyrosine improved sensitivity and accuracy by applying iqDBS. CONCLUSIONS: We established a new method for quantitative analysis of small molecules from dried blood spots that incorporates stable isotope internal standard at the time of blood collection.

[Determination of polychlorinated biphenyls in water by stable isotope dilution through automatic solid-phase extraction membrane coupled to gas chromatography-mass spectrometry].

OBJECTIVE: The method for analyzing of 20 polychlorinated biphenyls(PCBs) in water by stable isotope internal standard-automatic solid-phase extraction membrane(auto SPEM) coupled to gas chromatography-mass spectrometer(GC-MS) was established, and applied to the analysis of trace PCBs in water samples. METHODS: The sample backgrounds of bottled mineral water production in Fuzhou(groundwater), inland water in Fuzhou university city(surface water), tap water(pipe tip water) and stream water in Xiyuan Gong(water source of life) were analyzed by GC-MS. Surrogate solution(SS)internal standards including ~(13)C_(12 )PCBs stable isotope was used for quantification. The method was confirmed by calculating recovery rate(RR) and relative standard deviation(RSD) of spiked water samples(20, 40 and 80 ng/L, n=7). And recovery rates of SS were used to evaluate the effect of the pretreatment. And the method was used to determine PCBs of a total of 17 cross-sectiona river water in minjiang river tributaries upstream to the downstream estuary. RESULTS: The method detection limits(MDL) were 1. 9-6. 6 ng/L, and RRs were 70. 9%-127%, and RSDs were 0. 5%-13%. The RRs of SS were 40%-90%. Positive sample was not detected. CONCLUSION: The method of auto SPEM is less solvent, more automatic than traditional manual SPE. And the method is more accurate and reliable by stable isotope internal standard coupled to GC-MS. Therefore, it is useful for the trace PCBs determination of waters.

Quantification of cefazolin in serum and adipose tissue by ultra high performance liquid chromatography-Tandem mass spectrometry (UHPLC-MS/MS): application to a pilot study of obese women undergoing cesarean delivery.

Higher doses of cefazolin are required in obese patients for preoperative antibiotic prophylaxis, owing to its low lipophilicity. An ultra high performance liquid chromatography-tandem mass spectrometry method was developed to quantify cefazolin in serum and adipose tissue from 6 obese patients undergoing cesarean delivery, and using stable-isotope labeled cefazolin as an internal standard. The method has a 2µg/g lower limit of quantitation. The concentration in adipose tissue was 3.4±1.6µg/mL, which is less than half of the reported minimum inhibitory concentration of 8µg/mL for cefazolin. Serum cefazolin concentrations were more than 30-fold higher than in adipose tissue.