Ascorbic acid vitamin measurement

How analytical methods deal with interferences is one of the more ad hoc aspects of method validation. There is a variety of approaches to studying interference, from adding arbitrary amounts of a single interferent in the absence of the analyte to establish the response of the instrument to that species, to multivariate methods in which several interferents are added in a statistical protocol to reveal both main and interaction effects. The first question that needs to be answered is to what extent interferences are expected and how likely they are to affect the measurement. In testing blood for glucose by an enzyme electrode, other electroactive species that may be present are ascorbic acid (vitamin C), uric acid, and paracetamol (if this drug has been taken). However, electroactive metals (e.g., copper and silver) are unlikely to be present in blood in great quantities. Potentiometric membrane electrode sensors (ion selective electrodes), of which the pH electrode is the... 

Figure 5-5 shows data for an experiment in which ascorbic acid (vitamin C) was measured in orange juice by an electrochemical method. The current between a pair of electrodes immersed in the juice is proportional to the concentration of ascorbic acid. Eight standard additions increased the current from 1.78 to 5.82 p,A (column C), which is at the upper end of the desired range of 1.5- to 3-fold increase in analytical signal. 

One problem with glucose monitors is that species such as ascorbic acid (vitamin C), uric acid, and acetaminophen (Tylenol) found in blood can be oxidized at the same potential required to oxidize the mediator in Reaction 17-13. To correct for this interference, the test strip in Figure 17-10 has a second indicator electrode coated with mediator, but not with glucose oxidase. Interfering species that are reduced at electrode 1 are also reduced at electrode 2. The current due to glucose is the current at electrode 1 minus the current at electrode 2 (both measured with respect to the reference electrode). Now you see why the test strip has two working electrodes. 

Fumed silica A-200 (Pilot plant at the Institute of Surface Chemistry, Kalush, Ukraine specific surface area Ascorbic acid (vitamin C) and all-rac-a-Tocopheryl acetate (vitamin E acetate) were used as adsorbates. Folin-Ciocalteu s phenol reagent (Merck) was used to measure the total polyphenolic index. Silica samples with different degree of surface silylation were obtained using gas-phase chemical modification of highly disperse silica (A-200) surface by trimethylchlorosilane.6... 

Diced turkey bologna was dipped in water (none) or different antioxidants solutions before irradiated at 3 kGy. The antioxidants tested were 20 mM of ascorbic acid, vitamin E, and sesamol, or 0.75% of rosemary extract (rosemary). Volatile compounds were measured the next day, and expressed as square root of peak area count. The numbers were means of four replicates... 

Figure 20.4 Pulsed polarography. NPP and DPP techniques. The diagram shows at which instants the measurements are made (see the arrows a and h). Examples of measurements. The measurement of ascorbic acid (vitamin C) in a fruit juice (see conditions) corresponds to an oxidation on the working electrode.

The physicochemical properties of the water-soluble vitamins are extensively utilized in chemical methods. A method for quantitative vitamin C (ascorbic acid, AA) measurement in food and physiological samples is based on a reaction of the keto groups in dehydroascorbic acid (DHA) with o-phenylenediamine (OPD) to give a fluorescent quinoxaline. This method involves the oxidation of AA to DHA, followed by the measurement of total AA in the sample. The reductive capabilities of AA can especially be utilized for direct electrochemical (amperometric or coulometric) measurement when coupled with HPLC separation. 

This blood fraction contains white blood cells and platelets. Ascorbic acid (vitamin C) is carried mainly in the white blood cells and so its measurement in the buffy layer gives an indication of the vitamin C status of the body. 

However, this is true only for the sample solutions which contain a single component, H2O2 in this case. The electrochemical response is more complicated if sample solutions contain two or more redox species because both species would be oxidized or reduced, and thus they contribute concurrently to the output current. This is often the case for the electrochemical determination of drugs and metabolites or other biological components in blood because many kinds of redox-active species are intrinsically contaminated in blood. Ascorbic acid (vitamin C) and uric acid (UA) are redox species found in blood, and these compounds often disturb electrochemical measurements of blood because the redox potential... 

Food process optimi2ation measurements may link a single chemical such as a vitamin, or a physical change such as viscosity, to process conditions and to consumer acceptance. Retention levels of ascorbic acid [50-81-7] C HgO, or thiamine can often be used as an indicator of process conditions (see... 

Under stress conditions, such as cutting or light exposure, ascorbate oxidase has been described as promoting the transformation of ascorbic acid to dehydroascorbic acid (Wright and Kader 1997b). However, because ascorbic acid can be easily converted into dehydroascorbic acid, it is necessary to measure both ascorbic and dehydroascorbic acids to observe that the content of vitamin C was well preserved in fresh-cut fruit. 

The main value of ACU measurement is that it makes it possible to give a fast estimate of vitamin C deficiency because in healthy subjects ascorbic acid is about 90% of total ACU [33], A strong deficit of ACU, possibly due to vitamin C deficiency, was found in patients with breast tumors (Fig. 11). 

This experiment is flexible in that students may choose the samples they wish to analyze. It is suggested that several fruits and vegetables as well as an unknown ascorbic acid sample be available for analysis. The vitamin C content of raw and cooked vegetables can also be measured and compared. Commercially available vitamin C tablets or multivitamin pills provide interesting samples for analysis. The time required for this experiment can be adjusted by controlling the number and type of samples. 

B. Measurement of Vitamin C Standard Ascorbic Acid Solution... 

This is one vitamin that most laboratories can measure. There are a number of old-fashioned approaches that use 2,6-dichloroindophenol in a titrimetric method such as AOAC 985.33. This works well in some systems but can give rise to false positive results if there are other reducing substances present. It will not detect dehydroascorbic acid (DHA) and so it may well underestimate the actual vitamin C activity if a product contains a significant level of DHA. However, even with for these shortcomings, it is often used as a quick and rough method. In the AOAC there is also a fluorometric method (AOAC 984.26) where ascorbic acid is oxidised to DHA and this is reacted with o-phenylenediamine to give a fluorometric compound which can be detected. This is a robust method that has general applicability. 

A study on the photoprotective effect of the topical application of 2% vitamin E and 5% vitamin C in humans showed no effect with the application of each substance alone, but an enhanced photo-protective effect after applying vitamins E and C combined, which was attributed to the regeneration of vitamin E by vitamin C.20 This enhanced effect has also been shown with the topical application of a combination of 15% ascorbic acid and 1% a-tocopherol to porcine skin.21 The combined systemic supplementation of vitamins C and E was similarly able to reduce sunburn reactions22 and to increase the minimal erythema dose (MED), a measure for individual photosensitivity, more than supplementation with either vitamin E or vitamin C alone.23,24 An oral supplementation with an anti-oxidative combination of carotenoids (P-carotene and lycopene), vitamin C, vitamin E, selenium, and proanthocyanidins (Seresis , Pharmaton SA, Lugano, Switzerland) also reduced the development and grade of UVB-induced erythema 25... 

In reconstructed human epidermis the presence of vitamin C was required to normalize stratum corneum lipids, which was accompanied by an improvement of skin barrier formation.36 Interestingly the ascorbic acid concentration in the skin of atopic dermatitis37 and psoriatic patients38 measured in vivo by microdialysis was significantly lower than in healthy subjects. In psoriasis there was no significant difference in lesional versus nonlesional skin. There has also been demonstrated a decrease of ascorbic acid concentration in skin with increasing age.39... 

Lowering the pH is useful up to a point. It may favour the initiation of alternative mechanisms, but these tend to be less rapid. If the system contains ascorbic acid, then any measures taken to conserve vitamin C will help to prevent initiation of nonenzymic browning by its oxidation. Even the addition of extra ascorbic acid will prove useful for some time, but, once oxidised, browning will be worse than without it. 

Antioxidant activity of silica nanocomposites with immobilized vitamin C was tested using the polyphenolic activity index.8 After adsorption of ascorbic acid on the silica surface and centrifugation, the excess solution was removed to obtain the suspension of a fixed volume (2 ml). Distilled water, sodium carbonate solution, and Folin-Ciocalteu s phenol reagent were subsequently added to suspensions and to the reference Vitamin C solution. The suspensions were then stored for 30 min, and the optical density of supernatant was measured at X = 750 nm. The reference solution of ascorbic acid was used to compare antioxidant activity of vitamin-containing nanocomposites with the activity of dissolved vitamin C. 

A method of determining airborne iodine has also been reported.241 Here, iodine is absorbed into 5% aqueous KI and spectrophotometrically determined at 590 nm in the form of its complex with starch. This method is selective with respect to bromine and chlorine, and the sensitivity of this method is 0.25 mg of I2 per m3 of air. The concentration of the, 31I isotope in water can be determined by a method involving isotope exchange in the starch-iodine complex.242 Flow-injection determination of ascorbic acid (0.1-40 mg/mL) has been proposed.243 Iodine is generated in the flow system as I3- ions, which are in turn exposed to starch to produce a steady signal at 350 and 580 nm. Ascorbic acid provides inversed maxima which are measured. This method is recommended for analysis of ascorbic acid in fruit juice, jam, and vitamin-C preparations. Use of the blue complex has also been reported for determination of sodium dichloro-isocyanurate in air.244 Obviously the blue reaction is applicable in the determination of amylose, amylopectin, and starch,245-252 as well as modified starches.245,253-255... 

A coulometric titration method was introduced for sequential determination of sulfite, thiosulfate and ascorbic acid (1) in solutions containing sulfite-thiosulfate or sulfite-ascorbic acid couples. Formaldehyde or acetaldehyde can be used to mask the sulfite component. Two sequential measurements of coulometric time, one for both components in the mixture and one for the sample solution in which sulfite is masked, can be used to determine the concentrations of sutfite-thiosulfate and sulfite-ascorbic acid couples. The method is linear for 0.5-60 p,M 1 in the presence of 0.44-13 (xM sulfite, with RSD 0.1-4% and current efficiency of ca 98.0%. The method can be used for determination of the presence of sulfite and 1 in real sample matrices such as mineral waters and vitamin C tablets . 

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