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Bread contributes about one-sixth of our daily salt intake. Salt is used to improve flavor and texture. The amount of sodium in bread can differ significantly, depending upon brand and type of grain used, ranging from 592mg/100g to 748mg/100g. The level of sodium in bread is primarily due to the amount of salt added to the dough during manufacture. The amount of potassium is determined by the type of flour and the washing and bleaching operations during cereal processing. Bread does not make a significant contribution to maximum daily requirements for potassium. Method followed for this analysis is AOAC Official Method 990.23 (refer to BWB Method number 20). Procedure All solutions are prepared using deionized distilled water. A sample and pre-dried at 60ºC for 24 hours. The dried sample is then ground and homogenized in a suitable grinder. The test portion and crucible is weighed and dry ashed in an open inert vessel such as a crucible, then reweighed after ashing. Transfer the dried test sample to an airtight container of capacity about twice test portion size. Close container and mix sample thoroughly by shaking and inverting. Weigh sample to nearest 1mg, add a 1.25g portion of the sample into 50ml polypropylene beaker. Dissolve sample into approximately 20ml warm water while mixing. Quantitatively transfer contents to a volumetric flask and made up to volume. Alternatively a steam bath can be used to aid dissolution. Stock solutions are then made up of potassium and sodium; it is recommended that a single point calibration is carried out prior to examination to determine a rough estimated range in which the sample will fall. Once th is expected sample concentration is determined, at minimum 5 standard solutions should be produced for calibration of the instrument. The after calibration the sample can then be aspirated into the flame photometer, to decrease the likelihood of random errors, multiple samples should be aspirated. To obtain the concentration of sodium and potassium in the sample, it should be noted that a dilution step has been performed. The result of this experimental procedure is a value of concentration when dry ashed sample is diluted in 20ml water. Preparation of Standard Graph Set the flame photometer in accordance to MultiPoint/Single Ion Calibration found on page 24 of the BWB Technologies Installation and Operation Manual, to measure potassium emission. Nebulize the working standard solutions and adjust the controls until steady zero and maximum readings are obtained. Nebulize the intermediate working standard solutions and construct a graph relating raw emission data (known as RAW in BWB the flame photometer) to concentration of all the standard solutions. Measurement of Potassium and Sodium in Bread

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Measurement of Potassium in Unashed Plant Leaves 1 This procedure uses a TCA (trichloroacetic acid) extraction of the tissue to release all of the calcium. It was found that there was no significant interference from sodium, potassium or phosphate present in a sample at normal physiological levels. Calcium concentration in the range of 0.5 – 3.0 μg Ca / 100mg sample is readily determined. Procedure Solution and Standard Preparation Deionised distilled water is used throughout to prepare the following solutions: Organic Diluting Fluid containing 500ml isopropyl alcohol, 300 ml water, 0.2 ml Non-Ion-Ox (ex. Aloe Scientific Co., St. Louis, Mo. USA), 74.6 mg KCl and 292 mg NaCl. TCA solution for tissue extraction was 6.2% w/v and for standard preparation was 8.3% w/v and also contained 19.5 mg NaCl / 100ml. 6.25% trichloroacetic acid solution is first prepared. To do this accurately weigh out 6.25g of trichloroacetic acid and add it to a 100ml volumetric flask, then make up to the mark with deionised or milliq water. To make up 8.3% w/v TCA solution, 8.3g of TCA was dissolved in the same manner. Primary Calcium Standard Solution was prepared by dissolving 250 mg CaCO3 in 25ml of 2M HCl and diluted to 1l to give 100 μg Ca / ml Standards and reagent blanks containing 0 – 3 μg Ca /ml were prepared by mixing 80ml of Organic Diluting Fluid, 15ml of 8.3% TCA, 0 – 3ml of Primary Calcium Solution and deionised water to a final volume of 100ml. When carrying out analysis and calibration, you will be prompted to either analyse or skip blank analysis prior to calibration and sample measurement. The Organic Diluting Fluid prepared previously should be run as the blank for this analysis. Tissue Extraction Prepare a Slice of tissue approximately 0.5mm thick. Quickly dip the tissue slice in a calcium-free incubation solution and carefully blot on filter paper. Accurately weigh, to yield samples in the range of 100 mg wet weight. Transfer sample to centrifuge tube, add sufficient 6.2% TCA is give a volume of 2 ml. Vigorously shake the tube 3 times over a 30 minute period. Add a volume of 8 ml of Organic Diluting Fluid to the tube and centrifuge. The supernatant is analysed directly. Preparation of Standard Graph Set the flame photometer in accordance to MultiPoint/Single Ion Calibration found on page 24 of the BWB Technologies Installation and Operation Manual, to measure potassium emission. Nebulise the working standard solutions and adjust the controls until steady zero and maximum readings are obtained. Nebulise the intermediate working standard solutions and construct a graph relating raw emission data (known as RAW in BWB the flame photometer) to concentration of all the standard solutions. 1 Geyer, R.P. and Bowie, E.J., ‘The Direct Microdetermination of Tissue Calcium by Flame Photometry’, Anal. Biochem., 2, (1961), 360-369.

Measurement of Calcium in Biological Samples 1 A number of methods have been reported for the measurement of calcium in biological materials by flame photometry or titration. This method employs trichloroacetic acid (TCA) to precipitate the proteins and has been found to be adequate to extract all of the calcium from tissue samples. Sodium, potassium and phosphate caused no significant interference and calcium concentration in the range of 0 to 2 µg/ml is readily determined. Solutions Deionised distilled water is used throughout. Diluting Solutions is 500ml isopropyl alcohol, 300ml deionised water, 0.2ml Non-Ion-Ox (Aloe Scientific Co, St. Louis, Mo.), 74.6 mg KCl and 292 mg NaCl. Trichloroacetic acid solution is 6.2% w/v for tissue extraction. Procedure Tissue samples were prepared and carefully blotted on filter paper, then accurately weighed to yield samples of in the range of 100mg wet weight. The sample is transferred to a small centrifuge tube and sufficient 6.2% TCA solutions is added to bring the volume to 2 ml. The tube is tightly covered and vigorously shaken three times over a thirty minute period. A volume of 8 ml of Diluting Fluid is added to the centrifuge tube and the tube is centrifuged. The supernatant solution can be analysed directly without further dilution. When measuring samples, an initial test to estimate the range of calcium concentration via a single point calibration of 100ppm should be carried out. With the estimated value of sample concentration, standards should then be made up of at least 4 standards covering ±50ppm from the rough guide sample value. When producing standard solutions it is of vital importance that they are diluted in the Diluting Solution and the Diluting Solution should be ran as a blank so that the flame photometer accounts for chemical interferences in the matrix as well as the influence of isopropyl alcohol being present in the flame. Normal physiological concentrations of sodium, potassium and phosphate were not found to cause significant interferences. However methods for reducing the chemical interference found from these could be avoided by methods utilising EDTA, found in Measurement of Calcium in Serum and Urine. Phosphate interference can also be reduced by increasing flame temperature and methods have been proposed that the addition of trace quantities of Lanthanum and Strontium, however the extent by which the interference is reduced is not currently quantified. 1 Geyer, Robert & Bowie, E.‘The Direct Microdetermination of Tissue Calcium by Flame Photometry’,Anal Biochem, 2, (1961), p. 360-369.

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The BWB FLASH Photometer The newest feature to flame photometry. The BWB XP Flame Photometer Our Award winning 5- channel instrument! The XP is the first and only 5 channel flame photometer with simultaneous detection and display of all 5 elements. Find Out More New for 2020-2021 New for late 2020-2021 ‘ACT’ (Administrative Control Tool). This Unique BWB development provides data output controls with a Single Administrator password protected access code, and then 19 additional operator access passwords with a cascading access to the operation and calibration methods from greatest to least access. This controls both the on board Printer (ideal for a paper trail) as well as driving the AFHS (Automated Fluid Handling System) and the output from our Flame Photometer to clients Personal Computer or LIMS systems (FP-PC). This output allows for the creation of .csv formated Excel spreadsheets for sharing with Intranet or Internet multiple users. This ‘ACT’ has been developed at the request of our more sophisticated and demanding users embracing compliance standards of CFR 21 Part 11 and in Process Control QA, QC laboratories for stringent reporting requirements. For all new Generation 3 BWB Technologies Flame Photometers this is a no charge option at point of ordering. BWB Flame Photometers and Accessories Bringing you the best in Flame Emission Spectrometers and Accessories Go to link Go to link Go to link Go to link 1/1 View All Products Why choose BWB Flame Photometers? Simultaneous Detection Lithium, Sodium, Potassium, Calcium and Barium are measured simultaneously Advanced Calibration Stores calibration data and performs multi-mode calibrations -reducing analysis time by 65% Cost Savings The lowest "cost per use" and "cost of ownership" photometer in the industry. Animated Brochure View our online animated products brochure to read all specifications in detail. View Frequently Asked Questions Have questions about our products? Others might have had them too. Search through our FAQs section. View Installation, training, and scheduled maintenance available anywhere in the USA and Canada

Salinity in Processed Foods 1 Sodium chloride is an essential ingredient of all processed foods; it is needed to enhance flavour and increase shelf life. In many processed foods, additional sodium is added as a flavour enhancer; potassium and calcium are also naturally present but not normally used as processing additives. The optimum linear range for sodium measurement is within the range of 5 to 20ppm. Dilution of food samples to fall within this concentration range also causes the concentration of normally occurring levels of potassium and calcium to fall below an interfering threshold. High levels of calcium fortification should not interfere with sodium measurement (equivalent to 10g calcium per 100g of starting material). However, relatively high levels of potassium could interfere with the measurement of sodium. Procedure Samples of both plant and animal origin can be analysed; in cases where packing liquid and solid matter are packed together, the entire contents are homogenised. A 10g portion of the slurry is sampled and mixed with deionised double distilled water (40ml) and centrifuged (15,000g, 40C, 10 min). The supernatant was diluted and used for analysis. The resulting solution should be stored in polyethylene bottles prior to analysis. n.b. The bottles should be washed thoroughly and bathed in 5% HNO3 and rinsed twice with double distilled water prior to use. For more detailed procedures reference should be made to other applications on the www.BWBTech.com webpage under the applications tab. Preparation of Standard Graph Set the flame photometer in accordance to MultiPoint/Single Ion Calibration found on page 24 of the BWB Technologies Installation and Operation Manual, to measure potassium emission. Nebulise the working standard solutions and adjust the controls until steady zero and maximum readings are obtained. Nebulise the intermediate working standard solutions and construct a graph relating raw emission data (known as RAW in BWB the flame photometer) to concentration of all the standard solutions. 1 Chen, M-J, et al., Flame photometrica determination of salinity in processed foods’, Food Chemistry, 91 (2005) p. 765-770.

FP-PC and BWB Drive FP-PC App The BWB flame photometer has been configured to easily satisfy the most sophisticated IT requirements. The FP-PC app provides a useful interface to the BWB Flame Photometer (FP) facilitating the following operations: Normal calibration Report production Logging of streaming data for continuous on-line applications. (A 4-20 mA signal can also be output proportional to a calibrated channel on the FP). Extraction of data from the flame photometer when operated in automatic or semi-automatic mode with or without auto-sampler / auto-diluter known as Automatic Fluid Handling System (AFHS) Calibration manipulation including: 1. Upload from / download to the FP 2. Storage / retrieval of calibrations using the PC or system hard drive These functions are provided by the Welcome, Calibrate, Report, Logging, AFHS tabs together with one more (XP-Cal or BIO-Cal) that is dependent on the mode that the FP is being operated in. The FP-PC app, included with every purchase, has been written with GLP compliance in mind and facilitates the creation of data and reports in PDF format. BWB Drive New for 2014, BWB have created an industry leading USB flash drive packed full of features and resources designed to aide your daily analysis and BWB experience. Exclusively for BWB customers, this self-contained resource includes: Tutorial videos in 10 languages PC and Mac compatibility FP-PC app installation package (PC only) The XP Story – A short film about the technical wizards at BWB Links to BWB’s growing online video library (internet connection required) Manuals, guides, warranty information and registration PDF A list of international support and distributor contact details 13 GB of free space for you to use as you please! This wealth of information is provided in a beautifully designed BWB branded USB memory stick, which includes a built in flash light!

The BWB XP Flame Photometer Our award winning 5-channel instrument Ready to use straight out of the box! General Purpose Flame Photometer! T he BWB XP is the first and only 5 channel flame photometer with simultaneous detection and display of all 5 elements. The BWB XP was designed from the ground up, using new, innovative and modern technology to give unsurpassed levels of accuracy, usability and reliability while significantly reducing analysis time. Our customers do not need to waste valuable time changing filters and re-calibrating when a different measurement is required, as is a common requirement in all other Flame Photometer designs from 1951 to the present. With clean lines and contemporary bright colour schemes, the XP looks exceptionally modern and will fit into any laboratory. The instrument has a large LCD display, which can be viewed easily from any angle. Our metal casework provides a sturdy portable instrument and enables future recycling opportunities. ROHS compliant components and materials are used wherever possible. The BWB Carbon Footprint is exceptionally low, in keeping with corporate objectives of being a responsible manufacturer. Unlike other manufacturers who charge extra for vital accessories, BWB include everything required to use the product straight out of the box. Just add Gas!! (Propane, LPG or butane). We also provide as standard, data sharing via USB from the BWB XP to your PC. This ties the output of the BWB Flame Emission Spectrometer (FES) to the Internet or your company intranet systems and facilitates Distributed Control which is vital for Multi-National companies, Universities and various Governmental agencies. The BWB XP is the greatest advance in flame photometry for 30 years! FEATURES SIMULTANEOUS DETECTION AND DISPLAY OF ALL 5 ATOMS OF INTEREST INTUITIVE USER INTERFACE FOR TRUE EASE OF USE DISPLAY PROMPTS STEP BY STEP OPERATION BUILT-IN AIR COMPRESSOR SOLUTIONS AND LABWARE INCLUDED DATA SHARING VIA PC LINK VIA USB OPERATOR INDEPENDENT DETERMINATION OF RESULTS 5 USER SELECTABLE UNITS OF MEASURE USER SELECTABLE DECIMAL PLACES INTEGRATED PRINTER OPTION USES READILY AVAILABLE PAPER IQ, OQ, PQ WEB-BASED CERTIFICATION AVAILABLE: IQ OQ PQ is completely unique in the analytical instrumentation arena. Available in English, Russian, German, Mandarin, French and Spanish.More languages in development CORRECTION OF CA FOR THE INTERFERENCE FROM HIGH LEVELS OF NA Technical Data Sample rate 2 – 3.5ml/min Tubing materials Silicone and Tygon® Required desk space for Instrument 50cm (H) x 45cm (W) x 45cm (D)* For Safety reasons the Flame photometer requires 1m of unobstructed space above to allow dissipation of heat from the chimney. Instrument size XP Flame Photometer 51cm (H) x 38cm (W) x 41cm (D) (20in x 14in x 16in) Shipping 62cm (H) x 55cm (W) x 47cm (D) (24in x 22in x 19in) Weight XP Flame Photometer – 15.3kg (33.7lbs) Shipping – 25kg (55.1 lbs) Optimal range Single point calibration Na 0.1 – 60ppm K 0.05 – 100ppm Li 0.05 – 50ppm Ca 1.0 – 100ppm Ba 1.6 – 100ppm Multi Point Calibration Na 0.1 – 1000ppm K 0.05 – 1000ppm Li 0.05 – 1000ppm Ca 1.0 – 1000ppm Ba 5.0 – 3000ppm Reproducibility <1% Coefficient of variability for 20 consecutive samples over ten minutes at concentrations of 100ppm or less. (After instrument stabilisation). Specificity Na/K/Li = <0.5% to each other when equal in concentration at <100ppm Limit of detection (LOD) and limit of quantification (LOQ) LOD LOQ Na – 0.03ppm Na – 0.1ppm K – 0.02ppm K – 0.05ppm Li – 0.02ppm Li – 0.05ppm Ca – 0.3ppm Ca – 1ppm Ba – 1.6ppm Ba – 5.0ppm Time to stability Less than 15 seconds after sample is introduced into the flame. Ions measured Simultaneous measurement of Na, K, Li, Ca, Ba Interfaces USB 0-2.5 volt output (based on sample concentration linked to element of users choice) Optional 4-20mA output in place of the above Optional integrated printer .csv and .pdf generated reports and files via FP-PC software Recommended minimum warm up time based on ambient temperature 21ºC – 40 minutes Power requirements 100V – 250V AC at 50 or 60Hz automatically selecting Fuel requirement Propane, Butane or Natural Gas* regulated to 19Bar. Flow rate of 0.4l/min *with modifications. BWB Technologies recommends either Propane or Butane for optimum results. Readout LCD, four line, alpha numeric, back lit. WHAT’S IN THE BOX PC Leads USB and RS232 Selection of 3 Power cables to suit all regions Gas Hose Propane Adapter Aspiration Kit Manuals covering all aspects of the XP FP Quick Start guides Warranty Registration Form Certificates of analysis for all provided Fluids Material Safety Data Sheets for all provided Fluids The FP-PC Software installation USB 1L of BWB recommended cleaning solution 150ml calibration fluids for all 5 elements at 10,000ppm. 100ml conical flask with screw lid 100 disposable sample cups 1L of De-Ionised water 2m of waste tube Set of spare fuses 10 x 10ml pipettes 10 x 1ml pipettes

Determination of Sodium in Biodiesel by Flame Atomic This method allows the analyte to be pre-concentrated in the final solution, decreases the volume of strong acid used in the dissolution stage and does not use organic solvents. Procedure The sample of biodiesel is weighed into a crucible and placed in a heating muffle furnace. Two temperatures are used in the heating program for the dry decomposition: the first temperature (250ºC) is maintained for 1 hour to dry the sample. After this the temperature is progressively elevated to 600ºC and maintained for 4 hours to eliminate the remaining organic material. After cooling, the resulting inorganic residue is dissolved in a nitric acid solution (1.0% v/v). The resulting solution is transferred to a 100ml volumetric flask, and diluted to volume with dilute nitric acid (1.0% v/v). The LOD (limit of detection) for this method is given as 1.3 μg g-1 sodium and the LOQ (limit of quantitation) is 4.3 μg g-1 sodium. Preparation of Standard Graph Set the flame photometer in accordance to MultiPoint/Single Ion Calibration found on page 24 of the BWB Technologies Installation and Operation Manual, to measure potassium emission. Nebulize the working standard solutions and adjust the controls until steady zero and maximum readings are obtained. Nebulize the intermediate working standard solutions and construct a graph relating raw emission data (known as RAW in BWB the flame photometer) to concentration of all the standard solutions.