Milk, human chromatography

Schmid, D., Behnke, B., Metzger, J., and Kuhn, R., Nano-HPLC-mass spectrometry and MEKC for the analysis of oligosaccharides from human milk, Biomedical Chromatography 16(2), 151-156, 2002. 

Figure 10.13 GC clrromatogram obtained after on-line LC-GC(ECD) of a human milk sample analysed for PCBs (attenuation X 64). Peak identification is as follows (1) PCB 28 (2) PCB 118 (3) PCB 153 (4) PCB 138 (5) PCB 180 (6) PCB 170 (7) PCB 207. Reprinted from Journal of High Resolution Chromatography, 20, G. R. van der Hoff et al, Determination of organochlorine compounds in fatty matiices application of normal-phase LC clean-up coupled on-line to GC/ECD , pp. 222-226, 1997, with permission from Wiley-VCH.

When pushed to the limit by overriding human health concerns, residue chemists have achieved detection limits of Ippt (Ingkg ) or even into the low ppqr (1 pg kg ) range. An example at the 1 ppt level is provided by methods for 2,3,7,8-tetrachlorodibenzodioxin (TCDD) in milk and TCDD in adipose tissue. Eor relatively clean matrices such as water and air, preconcentration on solid-phase adsorbents followed by GC or gas chromatography/mass spectrometry (GC/MS) can provide detection limits of 1 ng m and less for air (examples in Majewski and Capel ) and 1 ngL and less for water (examples in Larson et A summary of units of weight and concentration used to express residue data is given in Table 1. 

Thurl, S., Offermanns, J., Muller-Werner, B., and Sawatzki, G., Determination of neutral oligosaccharide fractions from human milk by gel permeation chromatography, /. Chromatogr., 568, 291, 1991. 

Glausch, A., Hahn, J., Schurig, V. (1995). Enantioselective determination of chiral 2,2, 3,3,4,6 -hexachlorobiphenyl (PCB 132) in human milk samples by multidimensional gas chromatography/electron capture detection and by mass spectrometry. Chemosphere 30, 2079-2085. 

Miyazaki T, Yamagishi T, Matsumoto M. 1986. [Identification of 1,2,4,5-tetrabromobenzene and mirex in human milk by gas chromatography-mass spectrometry.] Journal of the Food Hygienic Society of Japan 27(3) 267-271. (Japanese)... 

Finke, B. Stahl, B. Pfenninger, A. Karas, M. Daniel, H. Sawatzki, G. Analysis of High-Molecular-Weight Oligosaccharides From Human Milk by Liquid Chromatography and MALDI-MS. Anal. Chem. 1999, 77,3755-3762. 

Analytical methods exist for measuring heptachlor, heptachlor epoxide, and/or their metabolites in various tissues (including adipose tissue), blood, human milk, urine, and feces. The common method used is gas chromatography (GC) coupled with electron capture detection (ECD) followed by identification using GC/mass spectrometry (MS). Since evidence indicates that heptachlor is metabolized to heptachlor epoxide in mammals, exposure to heptachlor is usually measured by determining levels of heptachlor epoxide in biological media. A summary of the detection methods used for various biological media is presented in Table 6-1. 

Heptachlor and heptachlor epoxide have been measured in samples of human milk using GC/ECD and GC/MS (Mussalo-Rauhamaa et al. 1988 Polishuk et al. 1977b Ritcey et al. 1972). Sample preparation steps for milk involve homogenization with chloroform/methanol, lipid extraction with petroleum ether, hexane or acetone-hexane, clean-up by column chromatography, and elution with acetonitrile, hexane, methylene chloride, or dichloromethane-petroleum ether. Precision, accuracy, and sensitivity were not reported for most of the studies however, one study reported a sensitivity in the low-ppb range (Ritcey et al. 1972). 

Wang H, Zhang J, Gao P, Yang Y, Duan H, Wu Y, Berset JD, Shao B (2011) Simultaneous analysis of synthetic musks and triclosan in human breast milk by gas chromatography tandem mass spectrometry. J Chromatogr B 879 1861-1869... 

Schwartz, H.E., Anzion, C.J.M., Van Vliet, H.P.M., Copius-Peereboom, J.W. Brinkman, U.A.T. (1979) Analysis of phthalate in sediments from Dutch rivers by means of high performance liquid chromatography. Int. J. environ, anal. Chem., 6, 133-134 Scott, R.C., Dugard, P.H., Ramsey, J.D. Rhodes, C. (1987) In vitro absorption of some o-phthalate diesters through human and rat skin. Environ. Health Perspect, 74, 223-227 Shaffer, C.B., Carpenter, C.P Smyth, H.F., Jr (1945) Acute and subacute toxicity of di(2-ethylhexyl) phthalate with note upon its metabolism. J. ind. Hyg. Toxicol., 27, 130-135 Sharman, M., Read, W.A., Castle, L. Gilbert, J. (1994) Levels of di-(2-ethylhexyl) phthalate and total phthalate esters in milk, cream, butter and cheese. Eood Addit Contam., 11, 375-385... 

Size exclusion chromatography coupled to an ICP-QMS with an octopole collision cell was employed for the multi-elemental speciation of essential elements (P, S, Cr, Mn, Fe, Co, Cu, Zn, Br, Se and I) and Al as a toxic element in premature human milk.28 Comparison of speciation results demonstrates that premature human mothers milk differs significantly from formula milks in terms of the element binding pattern to the biomolecules. It was found that premature human milk is very rich in high molecular weight species associated with metals. The authors concluded that more attention must be paid to the chemical form in which essential elements are added to the formulas, particularly those used for the nutrition of premature babies.28... 

Burger, R. L. and Allen, R. H. 1974. Characterization of vitamin Bi2-binding proteins isolated from human milk and saliva by affinity chromatography. J. Biol. Chem. 249, 7220-7227. 

Haroon, Y., Shearer, M. J., Rahim, S., Gunn, W. G., Mcenery, G. and Barkhan, P. 1982. The content of phylloquinone (vitamin Kd in human milk, cows milk and infant formula foods determined by high-performance liquid chromatography. J. Nutr. 112, 1105-1117. 

H Isshiki, Y Suzuki, A Yonekubo, H Hasegawa, Y Yamamoto. Determination of phylloquinone and menaquinone in human milk using high performance liquid chromatography. J Dairy Sci 71 627-632, 1988. 

B Zimmerli, R Dick. Determination of ochratoxin A at the ppt level in human blood, serum, milk and some foodstuffs by high-performance liquid chromatography with enhanced fluorescence detection and immunoaffinity column cleanup methodology and Swiss data. J Chrom B 666 85-99,1995. 

Y Pico, E Viana, G Font, J Manes. Determination of organochlorine pesticide content in human milk and infant formulas using solid phase extraction and capillary gas chromatography. J Agric Food Chem 43 1610-1615, 1995. 

MP Seymour, TM Jefferies, LJ Notarianni. An analysis of PCBs an organochlorine pesticides by capillary gas chromatography-a modern approach to PCB-OCP residue analysis of human milk. Anal Proc 23 260-261, 1986. 

MP Seymour, TM Jefferies, AJ Floyd, LJ Notarianni. Routine determination of organochlorine pesticides and polychlorinated diphenils in human milk using capillary gas chromatography-mass spectrometry. Analyst 112 427-431, 1987. 

The determination of TCDD in the final extract was by high resolution mass spectrometry multiple scan averaging. No gas chromatography was employed the sample was vaporized via the direct insertion probe (12.13). This method is inherently more prone to interferences than GC-MS (17.18) the response observed may not be due to TCDD. The results do serve as an indication, however, that if TCDD is present in human milk it is at 1 ppt or less. Thus any methods developed must be capable of this detection limit. 

Gas chromatography-low resolution mass spectrometry was used to determine if TCDD is in the samples (12). The GC column used was a 6 ft. x 2 mm i.d. glass 2.5% BMBT liquid crystal on 100/120 mesh Chromosorb WHP (Altech Assoc.) at 225°C. In Figure 3 is shown the output from a standard and typical human milk extract run on an LKB-9000 with a SI-150 (System Industries) data system equipped with a computer controlled multiple ion detector (Ledland, Inc.). The entire extract was... 

Figure 2. High Pressure reverse-phase liquid chromatography trace of 2,3,7,8-TCDD standard and a typical human milk extract...

Zarba A, Wild CP, Hall AJ, Montesano R, Hudson GJ, Groopman JD (1992) Aflatoxin Mi in human breast milk from The Gambia, west Africa, quantified by combined monoclonal antibody immunoaffinity chromatography and HPLC. Carcinogenesis, 13(5) 891-894. 

R. R. de la Flor St. Remy, M. L. Fernandez Sanchez, J. B. Lspez Sastre, A. Sanz-Medel, Multielemental distribution patterns in premature human milk whey and pre-term formula milk whey by size exclusion chromatography coupled to inductively coupled plasma mass spectrometry with octopole reaction cell, J. Anal. Atom. Spectrom., 19 (2004), 1104D1110. 

It has been shown that the major part of Ca and Mg are found in the soluble fraction (whey) of human milk, whereas only small amounts are present in insoluble caseins or in fat [11-13]. Calcium and Mg speciation studies in milk whey using size exclusion chromatography (SEC) combined with inductively coupled plasma atomic emission spectrometry (ICP-AES) [14] indicated that they were preferably associated with the milk nonprotein fraction. A further work [15] showed similar chromatographic profiles by SEC hyphenated with inductively coupled plasma mass spectrometry (ICP-MS), with Ca and Mg eluting in the lowmolecular-weight region (LMW) (<1.4 kDa). Thus, there seem to be weak associations, if any, of Ca2+ or Mg2+ with higher molecular mass biocompounds of milk. 

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