Quantified extracts

Standardized extracts" are herbal preparations adjusted within an acceptable tolerance to a given content of constituents with known therapeutic activity standardization is achieved by adjustment of the herbal preparations with inert material or by blending batches of herbal preparations. "Quantified extracts" are herbal preparations adjusted to a defined range of constituents adjustments are made by blending batches of herbal preparations. "Other extracts" are herbal preparations essentially defined by their production process and their specifications. 

Quantified extracts In the case of quantified extracts, the equivalent quantity or the ratio of the herbal substance to the herbal preparation shall be stated. Furthermore, content of the quantified substance(s) shall be specified in a range or... 

Phillips, F., and Privett, O. S. (1979). A simplified procedure for the quantified extraction of lipids from brain tissue. Lipids 14 590-595. 

This technique is used to quantify one or more components in a mixture, i.e., extracting them from mixtures to facilitate their final analysis. An example is that for the asphaltenes, already described in the definition of these components in article 1,2.1. 

Gc/ftir has both industrial and environmental appHcations. The flavor and aroma components in fragrances, flavorings, and foodstuffs can be identified and quantified via gc/ftir (see Food additives). Volatile contaminants in air, water, and soil can be analy2ed. Those in air are usually trapped in a sorption tube then injected into the chromatograph. Those in water or soil are sparged, extracted, or thermally desorbed, then trapped and injected (63,64). 

Quantitative mass spectrometry, also used for pharmaceutical appHcations, involves the use of isotopicaHy labeled internal standards for method calibration and the calculation of percent recoveries (9). Maximum sensitivity is obtained when the mass spectrometer is set to monitor only a few ions, which are characteristic of the target compounds to be quantified, a procedure known as the selected ion monitoring mode (sim). When chlorinated species are to be detected, then two ions from the isotopic envelope can be monitored, and confirmation of the target compound can be based not only on the gc retention time and the mass, but on the ratio of the two ion abundances being close to the theoretically expected value. The spectrometer cycles through the ions in the shortest possible time. This avoids compromising the chromatographic resolution of the gc, because even after extraction the sample contains many compounds in addition to the analyte. To increase sensitivity, some methods use sample concentration techniques. 

Purifications of elfamycins have been described in the Hterature using Craig distribution (2,34), chromatography on Sephadex LH-20 (2,14,26) and Amberlite XAD-2 (10,17,19,26), supercritical fluid extraction (37), and chromatography on an Ito multilayer cod planet centrifuge (26,38). and nmr assignments of most elfamycins have been accompHshed (3,24,26,32). The characteristic uv spectra permits some differentiation (12) and bathochromic shifts associated with Al " complexation have been used to quantify efrotomycin (2, R = CH ) in feed premixes (39,40). 

Analytical Techniques. Sorbic acid and potassium sorbate are assayed titrimetricaHy (51). The quantitative analysis of sorbic acid in food or beverages, which may require solvent extraction or steam distillation (52,53), employs various techniques. The two classical methods are both spectrophotometric (54—56). In the ultraviolet method, the prepared sample is acidified and the sorbic acid is measured at 250 260 nm. In the colorimetric method, the sorbic acid in the prepared sample is oxidized and then reacts with thiobarbituric acid the complex is measured at - 530 nm. Chromatographic techniques are also used for the analysis of sorbic acid. High pressure Hquid chromatography with ultraviolet detection is used to separate and quantify sorbic acid from other ultraviolet-absorbing species (57—59). Sorbic acid in food extracts is deterrnined by gas chromatography with flame ionization detection (60—62). 

An hplc assay was developed suitable for the analysis of enantiomers of ketoprofen (KT), a 2-arylpropionic acid nonsteroidal antiinflammatory dmg (NSAID), in plasma and urine (59). Following the addition of racemic fenprofen as internal standard (IS), plasma containing the KT enantiomers and IS was extracted by Hquid-Hquid extraction at an acidic pH. After evaporation of the organic layer, the dmg and IS were reconstituted in the mobile phase and injected onto the hplc column. The enantiomers were separated at ambient temperature on a commercially available 250 x 4.6 mm amylose carbamate-packed chiral column (chiral AD) with hexane—isopropyl alcohol—trifluoroacetic acid (80 19.9 0.1) as the mobile phase pumped at 1.0 mL/min. The enantiomers of KT were quantified by uv detection with the wavelength set at 254 nm. The assay allows direct quantitation of KT enantiomers in clinical studies in human plasma and urine after adrninistration of therapeutic doses. 

Prediction methods attempt to quantify the resistances to mass transfer in terms of the raffinate rate R and the extract rate E, per tower cross-sectional area Af, and the mass-transfer coefficient in the raffinate phase and the extract phase times the interfacial (droplet) mass-transfer area per volume of tower a [Eqs. (15-32) and (15-33)]. 

The ability to identify and quantify cyanobacterial toxins in animal and human clinical material following (suspected) intoxications or illnesses associated with contact with toxic cyanobacteria is an increasing requirement. The recoveries of anatoxin-a from animal stomach material and of microcystins from sheep rumen contents are relatively straightforward. However, the recovery of microcystin from liver and tissue samples cannot be expected to be complete without the application of proteolytic digestion and extraction procedures. This is likely because microcystins bind covalently to a cysteine residue in protein phosphatase. Unless an effective procedure is applied for the extraction of covalently bound microcystins (and nodiilarins), then a negative result in analysis cannot be taken to indicate the absence of toxins in clinical specimens. Furthermore, any positive result may be an underestimate of the true amount of microcystin in the material and would only represent free toxin, not bound to the protein phosphatases. Optimized procedures for the extraction of bound microcystins and nodiilarins from organ and tissue samples are needed. 

The usual means of identifying and quantifying the level of these additives in polymer samples is performed by dissolution of the polymer in a solvent, followed by precipitation of the material. The additives in turn remain in the Supernatant liquid. The different solubilites of the additives, high reactivity, low stability, low concentrations and possible co-precipitation with the polymer may pose problems and lead to inconclusive results. Another sample pretreatment method is the use of Soxhlet extraction and reconcentration before analysis, although this method is very time consuming, and is still limited by solubility dependence. Other approaches include the use of supercritical fluids to extract the additives from the polymer and Subsequent analysis of the extracts by microcolumn LC (2). 

One example of normal-phase liquid chromatography coupled to gas chromatography is the determination of alkylated, oxygenated and nitrated polycyclic aromatic compounds (PACs) in urban air particulate extracts (97). Since such extracts are very complex, LC-GC is the best possible separation technique. A quartz microfibre filter retains the particulate material and supercritical fluid extraction (SPE) with CO2 and a toluene modifier extracts the organic components from the dust particles. The final extract is then dissolved in -hexane and analysed by NPLC. The transfer at 100 p.1 min of different fractions to the GC system by an on-column interface enabled many PACs to be detected by an ion-trap detector. A flame ionization detector (PID) and a 350 p.1 loop interface was used to quantify the identified compounds. The experimental conditions employed are shown in Table 13.2. 

Pigure 13.16 shows the LC separation of this extract and the GC/PID chromatogram of the oxy-PAC fraction. The different oxy-PACs can be quantified without interfering compounds with a non-selective detector such as an PID. 

Poloxamers are used primarily in aqueous solution and may be quantified in the aqueous phase by the use of compleximetric methods. However, a major limitation is that these techniques are essentially only capable of quantifying alkylene oxide groups and are by no means selective for poloxamers. The basis of these methods is the formation of a complex between a metal ion and the oxygen atoms that form the ether linkages. Reaction of this complex with an anion leads to the formation of a salt that, after precipitation or extraction, may be used for quantitation. A method reported to be rapid, simple, and consistently reproducible [18] involves a two-phase titration, which eliminates interferences from anionic surfactants. The poloxamer is complexed with potassium ions in an alkaline aqueous solution and extracted into dichloromethane as an ion pair with the titrant, tet-rakis (4-fluorophenyl) borate. The end point is defined by a color change resulting from the complexation of the indicator, Victoria Blue B, with excess titrant. The Wickbold [19] method, widely used to determine nonionic surfactants, has been applied to poloxamer type surfactants 120]. Essentially the method involves the formation in the presence of barium ions of a complex be-... 

Adenylyl Cyclases. Table 5 Prodrug inhibition of [3H] cAMP formation in intact cells. Cells were prelabeled for 2 h with [3H]adenine before 50 pM forskolin and pronucleotides were added. After a 15 min incubation the newly formed [3H]cAMP was extracted and quantified as in (7)... 

Testing is undertaken by several methods, including chloroform extraction and use of a sulfonphthalein dye (absorbance of yellow-colored complex using bromophenol blue and bromocresol green) or the use of eosin (sodium tetrabromofluorescein) solution in acetone and tetrachloroethane solvent. After shaking with a citric acid buffer and eosin addition, upon standing the lower layer turns pink if filmer is present. Subsequent titration with Manoxol OT (sodium dioctyl sulfosuccinate) quantifies the filmer, with loss of the pink color indicating the end point. 

Figure 15-12 is a schematic illustration of a technique known as acid volatile sulfides/ simultaneously extracted metals analysis (AVS/SEM). Briefly, a strong acid is added to a sediment sample to release the sediment-associated sulfides, acid volatile sulfides, which are analyzed by a cold-acid purge-and-trap technique (e.g., Allen et ai, 1993). The assumption shown in Fig. 15-12 is that the sulfides are present in the sediments in the form of either FeS or MeS (a metal sulfide). In a parallel analysis, metals simultaneously released with the sulfides (the simultaneously extracted metals) are also quantified, for example, by graphite furnace atomic absorption spectrometry. Metals released during the acid attack are considered to be associated with the phases operationally defined as "exchangeable," "carbonate," "Fe and Mn oxides," "FeS," and "MeS."... 

To quantify the concentration of a colorant, one must consider that linearity between the colorant concentration and the fluorescence emission intensity exists only at very low concentrations. The reason for deviation from linearity may be reabsorption of the emission light by other fluorophores or formation of dimers. If no extraction and controlled dilution of the fluorescent colorant are performed, the colorant quantification will be only qualitative. 

Rios, A.O. and Mercadante, A.Z., Optimization of the conditions to obtain bixin crystals and for extraction and saponification to quantify bixin in extruded snacks by HPLC, AZi m. Nutr., 15, 203, 2004. 

Carmine extracted from cochineal insects is one of the most used natural colorings for beverages and other foods. Some representative articles refer to isolation and spectrometric analysis or the use of HPLC or capillary electrophoresis (CE) to separate and characterize all cochineal pigments. Its active ingredient, carminic acid, was quantified by rapid HPLC-DAD or fluorescence spectrometry. Carminic acid, used as an additive in milk beverages, was separated within 9 min using a high-efficiency CE separation at pH 10.0 after a previous polyamide column solid phase extraction (SPE), ... 

Five synthetic and five natural colorants were identified and quantified in lyo-philized dairy products and fatty foods using an automatic method based on solid phase extraction using a stationary phase followed by RP-HPLC C,g columns for the sequential retention of colorants and diode array detection. Lyophilization of the samples coupled with the separation procedure provided clean extracts despite the complexity of the food matrices and preserved the sample for at least 2 months without changes in colorant concentrations. The detection limits achieved for the colorants were found in a wide range from 0.03 to 75 pg/g of the lyophilized sample, according to the limits established by the European Union. ... 

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