SER 148 Extraction apparatus for the quantitative separation of a substance from a mixture by the use of an organic solvent Operation manual

Transcription

1 SER 148 Extraction apparatus for the quantitative separation of a substance from a mixture by the use of an organic solvent Operation manual IMPORTANT: READ THE INFORMATION CONTAINED IN THE PRESENT MANUAL BEFORE USING THE UNIT. THE MANUFACTURER DOES NOT ACKNOWLEDGE ANY RESPONSABILITY FOR AN IMPROPER USE OF THE EQUIPMENT, NOT RESPONDING TO DIRECTIONS FOR U SE. The labels applied to the unit warn the user on the dangers to which he is exposed during the use or the maintenance. The labels must be left on the unit and substituted if they are no more readable. Warn of danger Read carefully the Safety Regulations here below Caution hot suface! Do not dispose of this equipment as urban waste This unit must only be used for laboratory applications SAFETY RULES 1) During operation do not use nearby open flames or electrical equipment which could give raise to sparks in order to avoid fires. 2) The heating plate can reach a temperature of 260 C during the heating phase but it is still hot also during the cooling phase. Hang the heat shield before starting a cycle, to reduce risk. 3) The containers and the materials used during the work must be compatible with the temperature set on the unit. 4) Conform to the data of the table located on the console and presented in the operation manual which prescribes the temperature to be set for the different solvents and which sealing gasket is to be used. CLEANING Always unplug the unit before cleaning. The test tube must be cool. Use a damp cloth and not flammable, not aggressive detergents. PERSONAL PROTECTION EQUIPMENT The equipment used for personal protection must be compatible with the possible dangers deriving from the working materials and from the glass vessels. MAINTENANCE Maintenance by the user must be limited to the aspects described by the manual. Always unplug the unit before performing maintenance. According to the law on products guarantee our instrument must be repaired at our factory, save different agreements with our agents. GUARANTEE Starts from the date of delivery note and is referred to the number of register of the single unit /A11

2 Contents Forewords 1 Instrument description 2 Checking the instrument as received 2 Assembling and installing 3 Safety rules 4 Description of control panel 6 Programming and starting 7 Print out of the program s data 9 Choice of heating temperature 10 Flash point 12 Choice of seals 12 Operation cycle 14 Operations at the end of work 16 Maintenance 17 Oxidizing procedure for cleaning glassware 18 Electric scheme 19 Hydraulic scheme 21 Spare parts and accessories 23 Technical Features /A11

3 Analytical methods Determination of crude fat in meat and meat products 27 Determination of total fat in meat, fish, cacao and derived products 28 Determination of the total fat content in whole egg 29 Determination of fat content in vegetable proteic substances (oil seeds) 30 Extraction of oil and grease from wastewater 32 Extraction of oil and grease from sludges 33 Extraction of petroleum source rocks 34 Extraction of plasticizer mixtures from vinyl chloride plastics 35 Extraction of rosin from paper and paperboard 36 Extraction of oils from leather 37 Determination of unsaponified and unsaponifiable fat in soaps 38 Extraction of ethanol soluble components from detergent formulations 39 Extraction of finishing oils from textiles and synthetic or natural fibres 40 Organic analytes extraction from soil, sediment, sludges and waste solids in preparation for GC/MS, GC o HPCL analysis 41 Extraction of total fat from milk powder with a preliminary hydrolysis 43 Determination of total fat in cheese 44 Determination of total fat in flour, bread, bakery products and pasta with preliminary acid hydrolysis 45 Determination of oil content in fried potatoes 46 Determination of the fat content in animal feed 47 Determination of the total fat content in dry animal feed /A11

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5 Forewords Extraction by an organic solvent allows in many cases the quantitative separation of a substance or a group of substances (e.g. fat) from a mixture of solids or semisolids. The most widely used technique, all over the world, takes name from the German chemist Franz Soxhlet ( ) professor of chemistry at the High School of Agricolture of Munich (Bayern); it draws by distillation a pure solvent into contact with the mixture to be extracted. In the case of solid or semisolid mixtures the sample is contained in a porous cellulose or glass fiber extraction thimble which is covered by the pure solvent which drops from an overhanging condenser. From time to time the solvent with the extracted substances is drawn to the distillation vessel by an automatic siphon. The porous thimble retains the fraction of the mixture not soluble in the solvent. The dissolved substance must not be volatile at the boiling temperature of the solvent. The Soxhlet extraction apparatus has been modified in different aspects, some of which attempting to increase the temperature of solvent coming into contact with the mixture to be extracted, in order to reduce the extraction time. The Velp extraction apparatus uses a two stage procedure, in the first one the thimble to be extracted is directly immersed in the boiling solvent contained in the distillation vessel; after a defined time the thimble is removed from the solvent and hangs below the cooling condenser from which the condensed solvent drops and washes the sample. This procedure reduces the time required for a complete extraction to 1/4 1/6 of that required by a classical Soxhlet analysis. The fields of application include the analysis of food and feed, detergents, rubber and plastic formulations, pharmaceutical products, soil and so on, for soluble compounds as e.g. fats, tensides, plasticizers, pesticides. The Velp apparatus allows the use of many different solvents, a large fraction of which is recovered at the end of the extraction cycle, reducing to a minimum the related air pollution. The Velp apparatus is manufactured in accordance with the safety standard IP55 that allows a risk free operation. The manufacturer, engaged in a continuous improvement of his products reserves the right to modify any of their characteristics without notice /A11 1

6 Instrument description The unit is designed for the extraction of solid or semisolid materials using solvents according to Randall technique. The technique reduces sharply the extraction times using two phases with a final recovery of the used solvent. In the first phase the sample containing thimble is immersed in boiling solvent, while in the second phase it is removed from solvent and reflux washed. Finally a large fraction of the solvent is distilled and recovered. The unit with 3 or 6 places is optimized for manuality and safety. The chosen materials and technical innovation allow the respect of CEI EN norms with an IP55 degree of protection allowing a use safe of explosions with a limited space requirement. Checking the instrument as received When received, after packing removal check the integrity of unit. The instrument is delivered with 3/6 extraction cups, 3/6 Butyl seals, 3/6 extraction thimbles Ø 33 x 80 mm, 3/6 thimble adapters Ø 33 mm, tubes for hydraulic for water connection, a heat shield and an operation manual /A11 2

7 Assembling and installing Connect the instrument to a water tap for cooling water and to a water sink according to the autoadhesive labels on the rear of the instrument (C1): INLET The plastic tube is made fix by a band fastener. The open water tap must give a flow at the outlet of about 3 l per minute. DISCHARGE Cooling water to sink. C1 Connect the instrument to electric main 230 V (or 115 V) with connections for power and ground, protected by an earth leakage circuit breaker, tared for 0,5 A (C2). The connection to socket by cable and plug must be at least 1.5 m far from the instrument. Use the plug just installed at factory. The power capacity should be 10 A. It is not necessary that the equipment is placed in a forced ventilation environment (e.g. a fume hood). Do not use it without an adeguate air circulation to vent or in presence of points of danger like flames or electric sparks. C /A11 3

8 Safety rules Safety devices All electric components are protected according to IP 55 standards, assuring a complete separation from solvents. The equipment is protected against overheating by an electronic circuit with an indipendent safety temperature probe. Moreover there is a continuous cross control of the values given by the two temperature probes: when the two temperatures differ more than 20 C thermostatting is interrupted. The following cases can occur: 1. Interrupted temperature probe circuit. In this condition the temperature of the main probe raises to more than 350 C. If the interrupted circuit belongs to the safety probe, the display shows alternately AL2 and Short circuited probe. The apparent temperature of the probe is less than 2 C. For the main probe the display shows alternately AL1 and 000. If the safety probe is in short circuit the display shows alternately AL2 and Overheating. In this case the temperature of probes corresponds to the value of set point +20 C. For the main probe the display shows alternately AL1 and the temperature of heating plate. If the overheating is noticed by safety probe the display shows alternately AL2 and the temperature of the heating plate. 4. Lack of cooling water. A suitable flowmeter verifies that during operation there is no lack of cooling water necessary for the solvent vapour s condensation in the glass cooler. If the water mains is not open at the start of a new cycle of work, the cycle will not start, and an acoustic signal is triggered warning of the lack of water, and the visualizer alternatively indicates AL3 and H 2 O. If a lack of water occurs during the work cycle, the acoustic and visual alarm go off for one minute after which the cycle of work is automatically interrupted and AL3 remains on this display. To restart the corrected operation, press any key /A11 4

9 Recommendations for handling organic solvents As in general when handling chemicals, the following precautions should always be observed: a) Control the risk phrases ( R ) as hazard warning on the containers (e.g. R10 = flammable or R19 = may form explosive peroxides). b) Always protect eyes and possibly also skin, wearing glasses and gloves. c) Carry out work as much as possible in a well ventilated environment (e.g. fume hood). d) Wash immediately with plenty of water any splash on skin or eyes. If necessary seek medical attention. The supply of organic solvents in the laboratory should be reduced to an indispensable minimum. The solvents not in use should be preserved in metallic cabinets with holes, outside the laboratory. As far as possible it is recommended to avoid the use of most volatile solvents if they can be replaced by a less volatile homologous (e.g. toluene instead of benzene, heptane instead of pentane). Remember that ethers, unsaturated hydrocarbons, aldehydes and ketones are peroxide forming solvents and explosions can occur handling them not only by distillation but also by friction or violent agitation. The control of the presence of peroxides can be performed by easy to use test kits (e.g. Macherey-Nagel Quantofix peroxide test strips, Code CM and CM ). The destruction of peroxides is not always easy because two phases are formed with water solutions in most of cases which do not allow polymeric peroxides to come into contact with the used reducing agents. WARNING! The heating plate and the extract collecting vessels can reach a temperature as high as 260 C (set point). Use tongs to remove the extract collecting vessels at the end of extraction and solvent recovery phases. Disposal Dispose of in a manner consistent with the laws and regulations of the country and locality in which the unit will be disposed /A11 5

10 Description of control panel Main switch Display 1 (upper) Display 2 (lower) Leds Set key Key Key Key Key Start/Stop Key Air Supplies electric power to the unit Shows continuously the temperature of heating plate Shows all the settable parameter: Work programs: P01, P02. P29 Set temperature for each program. Operation times for each program (Immersion, Washing and Recovery). The turning on of corresponding led located on side of display warns about the correspondence of each value. The turning on of a led shows the correspondence of the value shown by the display: C Set temperature I Immersion time in boiling solvent W Reflux washing time R Solvent recovery time. Pressing key Set the letter P (of P01) flashes for a few seconds on the second display, referred to the used work program allowing the choice of a new program by the keys and. During programming conferms the value shown by the display. During a work cycle confirms the passage from a time to another. Increases the value shown by the second display. Decreases the value shown by the second display. By pressing this key the chosen program starts. The start of the cycle is warned by the turning on of the corresponding led. By pressing this key during a work cycle this is interrupted. Turns on the pump that introduces air in the water cooled condensers. The operation is warned by the turning on of the corresponding led /A11 6

11 Programming and starting Turn on the unit by pressing the lighted main switch located on control panel. The upper main display shows the temperature, as C, of the heating plate, while the lower display shows the number of the last set program. Choice of program: It is possible to set and choose among 29 work programs composed by: set temperature and three times developed successively with a warning by acoustic signal at the end of each (Immersion, Washing and Recovery). Pressing key Set, letter P flashes on the lower display; afterwards it is possible to select a new program with keys and. Setting of program: Each program is composed by four parameters: 1 Work temperature settable from 030 to 260 C 2 Immersion time settable from 000 to 999 minutes 3 Washing time settable from 000 to 999 minutes 4 Recovery time settable from 000 to 999 minutes Choose the work program to be set and then press the ky, the led C turns on and the lower display shows the set temperature, as C, which can be modified by the keys and from 030 to 260 C. Confirm the displayed value by the key, the led I turns on and the display shows the immersion time, in minutes, which can be modified by the keys and from 000 to 999 minutes. Confirm the displayed by the key and repeat the same procedure for the other two times (W Washing and R Recovery). After confirming the third time, the display shows the number of set program. If only the temperature of heating plate is to be set without the counting of time all the three times are to be setted to 000. If during the setting of a program no key is pressed, after 10 seconds the display returns to show the number of program. If a work cycle is developing it is not possible to modify the parameters of memorized programs nor those of the program in use. Pressing key brings the lower display to show in succession all the set parameters for the developing program /A11 7

12 Starting After choosing the desired work program, to start it press the key Start/Stop; the corresponding led will turn on showing the start of activity. The led C shows that the heating plate is reaching the set temperature, while the lower display shows the number of program in use. When the set boiling temperature is reached the lower display starts to show the decrease of immersion time, in minutes, while the led I (Immersion) turns on (knobs in Immersion position). At the end of immersion time (I Immersion) an acoustic signal warns abort the end of time. To start the count of washing time (W Washing) press the key, the led W turns on and the display shows the decrease of time (knobs in Washing position). At the end of washing time (W Washing) an acoustic signal warns about the end of time. To start the count of recovery time (R Recovery) press the key, the led R turns on and the display shows the decrease of time (closed stopcock = horizontal). At the end of the last time an acoustic signal warns about the end of cycle and the display shows end ; at the same time the power to heating plate is turned off. If it is wanted to move from one phase to the following without waiting the end of the set time it is sufficient to press the key. If the temperature of heating plate, at the start of a new cycle, is higher than the set value of the chosen program the cycle does not start with an acoustic warning and with the upper display showing EEE. Black-Out If during the development of a cycle a lack of electric supply occurs, the restoring of power restarts the equipment in the phase and time existing at the moment of interruption /A11 8

13 Print out of the program s data The instrument is provided with RS232 serial connection through a suitable socket in the front panel in order to print or save the data in files. So all information referring to a programme of work can be printed or saved on file. In addition to the cycle of work the following report can be printed by pressing the following keys at the same time: first and then SET The Print Report is as follows:: Solvent Extractor Version 1.5 Date:.. Time:.. Laboratory name:.. Operator Name: Sample name:... Lot number:... Program N : 01 Temperature ( C) 110 Immersion time (min.) 30 Washing time (min.) 20 Recover time (min.) 15 The optional accessories are the following: A A RS232 serial cable for the printer Printer The communication protocol of the serial socket is the following: Bits per second 4800 Data bits 8 Parity None Stop bit 1 Flow control No - Xon/Xoff If the print report is to be saved to the computer, the cable code A and the connector code A are necessary /A11 9

14 Choice of heating temperature The temperature of heating plate is to be chosen in relation to the used solvent (see Table). The difference between the temperature of heating plate and the boiling point of solvent varies from 100 C for those boiling at lower temperatures (e.g. diethyl ether, B.P C) to 260 C for those boiling at higher temperatures. This is due to the resistance to heat transmission presented by the glass of extraction vessels. It is to be noted that anyway the temperature of heating plate does never reach the ignition temperature of the used solvent. To avoid to overcome the boiling point, which gives rise to a tumultuous start of boiling, it is indispensable to add to extraction vessels some boiling chips before weighing for tare determination. Boiling chips (or bumping stones) = small spheres or pieces of glass, small pieces of pumice or silicon carbide. Temperature of heating plate to be set and type of seal required for some of the mostly used solvents: Solvent Glass cup Recommended Temperature C Recommended Seal Set Boiling Point C Ignition Point C Acetone 160 Butyl Chloroform 160 Viton Not combustible Diethyl ether 110 Butyl Ethanol 210 Butyl Ethyl acetate 190 Butyl Methanol 210 Viton/Butyl n-hexane 130 Viton Petroleum ether 110 Viton Not specified Toluene 210 Viton Trichloroethylene 200 Viton 86.7 Not combustible If the analysis takes too long the recommended temperature for each solvent can be increased by about 20% /A11 10

15 Commonly used solvents listed by increasing boiling point. SOLVENT BOILING POINT C FLASH POINT C AUTO IGNITION TEMPERATURE C DENSITY g/ml at 20 C VAPOUR PRESSURE mm Hg a 25 C SPECIFIC HEAT gcal/g/ C EVAPORATION SPECIFIC HEAT gcal/g Petroleum ether >200 C >400 ~0.500 ~80 Diethyl ether Pentane (C) Methylene chloride (605) (*) Acetone Chloroform (*) Methyl alcohol Tetrahydrofuran n-exane Carbon tetrachloride (*) Ethyl acetate Ethyl alcohol Methyl ethyl ketone Benzene (1) Cyclo hexane Acetonitrile Isopropyl alcohol Trichloroethylene (*) n-heptane Isoctane Dioxane Isobutyl alcohol Toluene n-butyl alcohol Acetic acid Octane n-amyl alcohol (C) Xilene (*) not flammable (C) closed cup (1) definitely known to be cancinogenic /A11 11

16 Flash point The temperature at which a liquid gives off a quantity of vapour sufficient to form an ignitable mixture with the air near the surface of the liquid or within the test vessel. There are two groups of experimental devices for flash point determination. For the purposes of the official shipping regulations (ICC, OG, IATA) the flash point is determined by the Tagliabue open cup method (ASTM D-1310/63) usually abbreviated TOC. IATA also admits the Abel or the Abel-Pensky closed cup tester. Other methods used, generally for the higher flash points, are the Tag closed cup and Cleveland open cup. The open cup methods approximate better actual conditions. Choice of seals The building materials of retaining seals located between the extraction vessels and the upper parts of the equipment (water cooled condensers) ask for a particular attention due to their chemical resistance to different solvents. Two materials were chosen: Viton (fluorinated polymer) and Butyl rubber (synthetic rubber) both provided with the instrument. In general terms Viton is easily dissolved by chetones and esthers, while butyl rubber is attacked by hydrocarbons and halogenated solvents. Before starting an extraciton it is to be controlled the compatibility of seals for the used solvent (see Table page 12). Butyl rubber seal WHITE Viton seal BLACK /A11 12

17 It s available, on request, the Vaflon retaining seal (particular elastomer), resistant to all types of solvents used in extractions, for applications that need particular solvents (or a combination of solvents that would need separately the use of different gaskets). The Vaflon seal and the Butyl seal are white. The butyl seal shows a little sign on the external diameter that helps to mark the two materials. Vaflon seal WHITE A Butyl rubber seal WHITE /A11 13

18 Operation cycle After the installation of the new equipment it is suggested to perform a blank cycle of work with a solvent. This is aimed to remove traces of grease deriving from the assembling which could impair the results of first extractions. 1. Turn on the unit by the general switch located on control panel. 2. Open cooling water tap. 3. Set the temperature of heating plate in relation to the used solvent (see Table page 12). Verify the compatibility of retaining seals. As delivered the unit is equipped by Viton seals. If the used solvent requires Butyl seals proceed to change as described in Maintenance. 4. Introduce samples into thimbles. Connect thimbles to thimble adapters (C3) wearing gloves to avoid contamination (fat). Introduce thimbles with thimble adapters into the unit (C4) with front knobs in Immersion position. Then place the knobs in Washing position (C5). 5. Place extraction vessels, preweighted with boiling chips and filled with solvent, into position on the heating plate (C6). 6. Pull down the lever retaining firmly the extraction vessels. 7. Place the glass tap located on condensers in vertical position (open) (C7). 8. Place front knobs in Immersion position. 9. Start heating. 10. At the end of extraction in boiling solvent, move the knobs in Washing position. 11. At the end of reflux washing, place the glass tap located on condensers in horizontal position (closed) and wait for solvent recovery into lower part of glass condenser. 12. Disconnect the retaining lever and remove glass extraction vessels. Remove extraction thimbles and thimble connectors. Recover solvent placing empty beakers on the heating plate and opening the stopcocks. C3 C4 C /A11 14

19 C6 C7 Randall system Extraction Washing Recovery /A11 15

20 Operations at the end of work At the end of instrument use: 1. Close the cooling water tap (A6). 2 Turn off the unit by the general switch. Leave always empty glass beackers on the heating plate to avoid dripping. A /A11 16

21 Maintenance Cleaning of cooling tap water filter Unplug the unit from electric supply and remove the connection to tap water. Remove the cooling water tube from the unit and pull out the filter using small pincers. Clean the filter under flowing water or by compressed air (A11). Replacement of seals Retaining seals between extraction vessels and glass condensers are to be changed if required by the used solvent. See the table located on the unit or in the present operation manual. Anyway seals are to be replaced if wearied or no more elastic. Get hold of a seal by a small pincer or screwdriver and pull downward (C9). A11 C /A11 17

22 Oxidizing procedure for cleaning glassware Glassware can be washed normally using a solution prepared dissolving 30 g of sodium hydroxide (NaOH, technical grade), 10 g of sodium tripolyphosphate (Na 5 P 3 O 10, technical grade) and 2 g of sodium lauryl sulfate (C 12 H 25 NaSO 4, commercial mixture). Glassware can be cleaned by chromic acid oxidation (dichromate and concentrated sulfuric acid), sulfonitric acid oxidation (warm concentrated sulfuric acid with small amounts of potassium or sodium nitrate or nitric acid), or by nitrous oxide oxidation (sodium nitrite and concentrated sulfuric or hydrochloric acids). Chromic-sulfuric acid mixture is prepared adding slowly 1 l of concentrated sulfuric acid (H 2 SO 4, d.1.84, technical grade) to 35 ml of saturated sodium dichromate acqueous solution (Na 2 Cr 2 O 7, 73% w/w at 20 C, technical grade) while stirring. The cold mixture must be left into contact with glassware to be cleaned at least 15 minutes, better overnight. The freshly prepared solution is orange red and becomes green after repeated use due to the reduction of Cr VI to Cr III. Exausted chromic-sulfuric mixture must be disposed of in a proper manner due to environmental toxicity of chromium. Nitrous oxide oxidation of organic residues can be obtained washing the glassware with hot concentrated sulfuric acid (H 2 SO 4, 70%, d technical grade) to which a few drops of saturated sodium nitrite acqueous solution (NaNO 2, technical grade, 100 g into 150 ml of tap water) are added before use. Another method is based on concentrated nitric acid oxidation: the addition of a few drops of ethanol produces after minutes an exotermic reaction and the evolution of oxidizing nitrous oxide. Attention: all these treatments must be performed under a fume hood due to the extreme toxicity of evolved nitric-nitrous fumes. After oxidizing treatment the filters must be thoroughly rinsed with tap water and finally with deionized water and then dried. Using nitric acid protect hands with PVC gloves and not rubber which is attacked. Rusting of components Chlorinated hydrocarbons contain and release on heating traces of very aggressive hydrochloric acid. As a consequence thimble holders and thimble adapters can show some rusting. The very special steel used for these component is slightly attaked by hydrochloric acid. The rust can be easily removed by immersion of the components in a solution composed by nitric acid, technical grade, concentrated, 2 volumes; hydrochloric acid, technical grade, concentrated, 1 volume; tap water, 5 volumes. By heating the solution to C the rust is removed in a few minutes. After rinsing dry the components /A11 18

23 Electric scheme 230 V/50 Hz 1) General lighted switch 2) Electric board 3) Air pump 4) Heating plate 5) Working Pt100 probe 6) Safety Pt100 probe 7) Male connector 8) Female connector 9) Flow rate 10) RS232 socket /A11 19

24 Electric scheme 115 V/60 Hz 1) General lighted switch 2) Electric board 3) Air pump 4) Heating plate 5) Working Pt100 probe 6) Safety Pt100 probe 7) Male connector 8) Female connector 9) Flow rate 10) RS232 socket 11) Transformer /A11 20

25 Hydraulic scheme Ser 148/3 N Material Diameter Lenght Q.ty (mm) (cm) 1 EPDM 6.4x EPDM 6.4x EPDM 6.4x MQ/MVQ 3x MQ/MVQ 3x EPDM = Butyl MQ/MVQ = Silicon /A11 21

26 Hydraulic scheme Ser 148/6 N Material Diameter Lenght Q.ty (mm) (cm) 1 EPDM 6.4x EPDM 6.4x EPDM 6.4x EPDM 6.4x EPDM 6.4x MQ/MVQ 3x MQ/MVQ 3x MQ/MVQ 3x EPDM = Butyl MQ/MVQ = Silicon /A11 22

27 Spare parts Cod. Art. Cod. Art. Description: 230V/50 Hz 115V/60 Hz Electronic box IP Heating plate IP 55 (SER148/3) Heating plate IP 55 (SER148/6) Air pump IP 55 Accessories Cod. Art. 230V/50 Hz Description: A Extraction cups 6 pcs/box A Thimble connector Ø 33 mm Viton retaining seal Butyl rubber retaining seal A Handling device for extraction cups A Thimble weighing cup A Pincer for weighing cups CM Pure cellulose extraction thimbles Ø 33x80 mm A Thimbles stand (6 places) Water cooled glass condenser A Vaflon retaining seal /A11 23

28 Technical features SER148/3 General F Power supply V/Hz 230/ /50-60 Total electric power W 500 Dimensions (WxHxD) mm 480 x 620 x 390 Inch 18,9x24,4x15.5 Weight kg 30 Lb 66 Display 2 display with 3 digit Pollution degree CEI EN Measurement category CEI EN Max. admitted humidity T % 85 Storage temperature range C +5 C +60 C F Operating temperature range C +5 C +40 C F Fuse (T) A 5 10 Performances Reproducibility (RSD) % 1 Solvent recovery % from 50 to 75 Consumption Cooling water lt/1 2 Sample volume g from 0.5 to 15 Solvent volume ml from 30 to 100 for each position Programs Number of programs N 29 Settable parameters and values: Working temperature from 30 to 260 C Immersion time from 0 to 999 minutes Washing time from 0 to 999 minutes Recovery time from 0 to 999 minutes Safety International protection (EN 60529) Over temperature Cooling water IP55 2 internal circuit with 2 separate probe Alarm for absence or insufficiency of cooling water /A11 24

29 Technical features SER148/6 GENERAL F F Power supply V/Hz 230/ /50-60 Total electric power W 950 Dimensions (WxHxD) mm 700 x 620 x 390 Inch 27,5x24,4x15.5 Weight kg 40 Lb 88 Display 2 display with 3 digit Pollution degree CEI EN Measurement category CEI EN Max. admitted humidity T % 85 Storage temperature range C +5 C +60 C F Operating temperature range C +5 C +40 C F Fuse (T) A 5 10 Performances Reproducibility (RSD) % 1 Solvent recovery % from 50 to 75 Consumption Cooling water lt/1 2 Sample volume g from 0.5 to 15 Solvent volume ml from 30 to 100 for each position Programs Number of programs N 29 Settable parameters and values: Working temperature from 30 to 260 C Immersion time from 0 to 999 minutes Washing time from 0 to 999 minutes Recovery time from 0 to 999 minutes Safety International protection (EN 60529) Over temperature Cooling water IP55 2 internal circuit with 2 separate probe Alarm for absence or insufficiency of cooling water /A11 25

30 Flow sheet for a typical extraction of soluble matter by an organic solvent from solid samples A B C D E F G Grind and homogenize the sample Weigh the sample, from 3 to 10 g in relation to the presumed content of extractable matter When vegetable or animal protein containing products are examined a preliminary hydrolysis with hydrochloric acid should be performed. Boil one hour in a flask and separate the residue on a paper filter Transfer quantitatively the sample or the residue from hydrolysis with the paper filter into the extraction thimble and cover it with a defatted cotton plug Introduce the thimble into the extraction equipment Weigh the extract collecting vessel with some boiling stones, beforehand dried to constant weight, and introduce it into the extraction equipment Add the solvent ( ml) H Close the extraction unit, start the cooling water flow and the heating. When the solvent starts boiling immerse the thimble by placing the slider into Immersion position I L M After minutes extract the thimble from solvent placing the slider into Washing position After minutes of reflux washing close the stopcock located under the water cooled condenser and evaporate completely the solvent from the extraction vessel. If the used solvent has heavy vapour turn on the air pump to improve the displacement to the condenser Dry the extract collecting vessel with boiling stones in an oven during 30 minutes at a temperature high enough to evaporate completey solvent residues N O Cool the extract collecting vessel in a dessicator and weigh it Compute the result. The weight of extract is given by the weight of point N less the weight of extract collecting vessel with boiling stones of point F and is to be referred to sample weight as point B /A11 26

31 Determination of crude fat in meat and meat products The solvent extraction is performed on dried samples. 1) Sample: 3 g of grounded and homogenized material is weighed in an extraction thimble with 1 mg proximation. Total weight-thimble weight = wet sample weight. 2) Chemicals: Petroleum ether C, with an evaporation residue not higher than 10 mg/l. Sand: Sea sand or siliceous sand with an extractable content lower than 4 mg/ 5 g. Cotton: defatted cotton wadding Boiling stones. 3) Sample drying: Add some sand to the weighed sample in thimble and mix with a glass rod. Clean the rod with a small flock of defatted cotton and add this to thimble. Dry the thimble 1 hour in a 125 C oven. Remove the thimble from oven and let it cool in a desiccator. Loosen the sample and sand mixture with a glass rod. Clean the rod with a small flock of defatted cotton and place this on top of thimble. 4) Extraction: Accurately weigh the glass extraction cup with a few boiling stones (1 mg proximation). Add 40 ml of petroleum ether. Transfer thimble with sample and sand, assembled with thimble connector to extraction unit. 30 minutes with thimble immersed in boiling solvent 60 minutes of reflux washing 5) Weighing: Dry the extract containing cup with boiling stones in an oven at 125 C during 30 minutes. Let it cool in a desiccator. Weigh to 1 mg proximation. 6) Expression of results: Refer to wet sample as weighed and give the results as crude fat/100 g with two decimal figures. 7) Reference: AOAC. Official Methods of Analysis (1990) 15th Ed. 3rd Supplement Arlington, Virginia, USA /A11 27

32 Determination of total fat in meat, fish, cacao and derived products A preliminary acid hydrolysis is aimed to free occluded or bound lipids 1) Sample: 2 to 10 g of grinded and homogenized matter, weighed in a 250 ml Erlenmeyer flask (with reflux condenser) with 1 mg proximation 2) Chemicals: Hydrochloric acid (HCl) about 4 N, 100 ml of concentrated hydrochloric acid, reagent grade, d. 1.19, mixed with 200 ml of water. Exane or petroleum ether C with an evaporation residue not higher than 10 mg/l. Boiling stones. 3) Sample hydrolysis: Add 100 ml of 4 N hydrochloric acid to Erlenmeyer flask with sample. Mix well and after assembling the reflux condenser boil 1 hour, add 100 ml of hot water and filter on a double folded paper filter. Wash three times the Erlenmeyer flask with ml of hot water and add the washings to the double filter with hydrolyzed sample. Wash with water until no more acid reaction is shown by filtrate. 4) Sample drying: Place the double filter paper in a 300 ml beaker and place 1-2 hours in an oven at 103 C, together with the Erlenmeyer flask used for the hydrolysis. The dry filters are rolled and introduced in an extraction thimble. The internal of Erlenmeyer flask and beaker are cleaned with a cotton plug soaked in solvent, using forceps. The plug is added to thimble. 5) Extraction: ml of the chosen solvent 30 minutes with thimble immersed in boiling solvent 60 minutes of reflux washing Boiling chips 6) Extract drying: Extraction vessel with boiling stones in an oven at 103 C for 30 minutes. Cool in a desiccator. 7) Weighing: The extract containing vessel with boiling stones is weighed to 1 mg proximation after 30 minutes of heating in an oven at 103 C. 8) Expression of result: Refer to wet sample as weighed and give the result as fat g/100 g with one decimal figure 9) Reference: Amtliche Sammlung von Untersuchungsverfahren (1982). (Official Collection of German Analytical Procedures). L Bestimmung des Gesamtfettgehaltes (Determination of total fat content) /A11 28

33 Determination of the total fat content in whole egg 1) Sample: 5-10g of liquid pasteurized egg are weighed into an Erlenmeyer flask connectable to reflux condenser. 2) Chemicals: hydrochloric acid (HCl) 4N: 100ml of concentrated hydrochloric acid, d.1.19, mixed together with 200ml of deionized water; chloroform (temperature for the heating plate: 160 C) or 1:1 mixture toluene/ethanol (temperature for the heating plate: 210 C), reagent grade. 3) Sample hydrolysis: from 50 (5g sample) up to 100ml (10g sample) of 4N hydrochloric acid. After assembling the water cooled reflux condenser the Erlenmeyer flask is heated to boiling for one hour.at the end of hydrolysis 150ml of deionized water are added to the sample. 4) Sample filtration: a double paper filter soaked with water is used to filter the hydrolyzed sample (Whatman n.o. 42 filtering disks). The hydrolyzed sample is to be cooled at C about (using an ice bath) before filtration. The flask used to hydrolyze the sample is to be washed using deionized cool water and the washings are to be poured on the filters. Wash by using cool water until the washings don t show acid reaction to litmus paper (500ml of washing water are required about). 5) Sample drying: The double paper filter is placed on a watch glass and desiccated in an oven at 102 C for 2 hours together with the flask used for the hydrolysis. The dry filter is rolled and introduced into an extraction thimble. The internal part of flask and watch glass are wiped with filter paper soaked in solvent which is also introduced into thimble. Use forceps. 6) Extraction: 60-70ml of the chosen solvent (according to sample weight). 15 minutes with thimble immersed into boiling solvent 30 minutes of reflux washing 7) Extract drying: place the extraction vessel in an oven at 100 C for 30 minutes. Let cool in a desiccator up to room temperature. 8) Weighing: the extract containing vessel is weighed with a tolerance margin of 1mg. 9) Expression of the results: % of fat = W3 W2 / W1 X 100 W1 = sample weight (g); W2 = extraction vessel weight (tare,g); W3 = vessel weight+ residue weight (g). 10) Reference: AOAC. Official Methods of Analysis. Arlington, Virginia, USA, method /A11 29

34 Determination of fat content in vegetable proteic substances (oil seeds) Fat content is defined as the fraction extractable with a solvent in the conditions suggested for the utilized method. Occluded or bound lipids are freed before extraction by acid hydrolysis. 1) Sample: 5 to 10 g of product grinded to less than 1 mm size, weighed in a 250 ml Erlenmeyer flask (with reflux condenser). 2) Chemicals: Hydrochloric acid (HCl), about 4 N, 100 ml of concentrated hydrochloric acid, d. 1.19, mixed with 200 ml of water. Diatomaceous flour as filter aid. Exane or petroleum ether C with an evaporation residue not higher than 10 mg/l. Boiling stones. 3) Sample hydrolysis: From 50 (for a 5 g sample) to 100 ml (for a 10 g sample) of 4 N hydrochloric acid. After assembling the water cooled reflux condenser the Erlenmeyer flask is heated to boiling for one hour. At the end of hydrolysis 150 ml of water and about 1 g of filter aid are added to the sample still hot. 4) Sample filtration: A double paper filter soaked with water is used to filter the hydrolyzed sample. The Erlenmeyer flask used for the hydrolysis is washed with hot water and the washings are poured on the filter. Wash with water until the washings do not show acid reaction to turmeric indicator paper. 5) Sample drying: The double paper filter is placed on a watch glass and essiccated in an oven at 102 C for 2 hours together with the flask used for the hydrolysis. The dry filters are rolled and introduced in an extraction thimble. The internal of flask and watch glass are wiped with filter paper soaked in solvent which is also introduced in thimble. Use forceps. 6) Extraction: ml of the chosen solvent (according to sample weight) Boiling stones 15 minutes with thimble immersed in boiling solvent 30 minutes of reflux washing. 7) Extract drying: Extraction vessel with boiling stones in an oven at 100 C for 30 minutes. Cool in a desiccator. 8) Weighing: The extract containing vessel with boiling stones is weighed to 1 mg proximation. 9) Control of extraction: Repeating the extraction on the extracted sample in the same conditions should give an extract of less than 2 mg; if not, use longer extraction time /A11 30

35 10) Expression of result: As % of fat, with 2 decimal figures, stating which solvent was used. Fat g of extract % g of sample x ) Reference: Experimental Station for oil and fat industries (1988). Determination of fat. Method N 4. Milano, Italy /A11 31

36 Extraction of oil and grease from wastewater The method determines groups of substances with similar physical characteristics on the basis of their common solubility in carbon tetrachloride. Time ago the used solvent was a freon (trichlorotrifluoroethane) dismissed for ecological reasons. Compounds volatilized at or below 103 C will be lost when the filter is dried. The method is empirical andd reproducible results are obtained only following exactly all details. 1) Sample: 1 l of wastewater collected in a wide mouth glass bottle and immediately acidified to ph 2 or lower with hydrochloric acid (HCl) to hydrolyze soluble soaps. 2) Chemicals: Carbon tetrachloride (CCl 4 ), reagent grade, with an evaporation residual not higher than 5 mg/l (0.005 ). Distilled water Diatomaceous flour filter aid suspension 10 g/l in distilled water Boiling stones 3) Sample filtration: Prepare a 12 cm Buchner funnel with a disk of muslin cloth overlaid with a filter paper disk. Filter under vacuum 100 ml of filter aid suspension on the Buchner and wash with 1 l of distilled water. Filter acidified sample until no more water passes the filter. Transfer paper filter to a watch glass using forceps. Wipe walls and bottom of collecting bottle and Buchner funnel with pieces of filter paper soaked in solvent and add them to paper filter on watch glass. Roll the paper filter and introduce it into extraction thimble. Dry thimble in an oven at 103 C for half an hour. 4) Extraction: ml of solvent Boiling stones 15 minutes with the sample immersed in solvent 30 minutes of reflux 5) Extract drying: Extraction vessel in an oven at 80 C for 15 minutes. Cool in a desiccator. 6) Weighing: The extract containing vessel with boiling stones is weighed to 0.1 mg proximation. 7) Expression of result: As mg/l with one decimal figure 8) Reference: APHA-AWWA-WPCF (1985) Standard Methods for the Examination of Water and Wastewater. 16th Ed. Washington D.C., USA /A11 32

37 Extraction of oil and grease from sludges Acidified sludge essiccated by heating gives low results. Drying is performed mixing the sludge with magnesium sulfate monohydrate which combines with water for a 75% of its weight giving the eptahydrate. 1) Sample: About 200 g of wet sludge collected in a wide mouth glass bottle. Weigh 20 ± 0.5 g of sample in a 150 ml beaker. Acidify to ph 2.0 with hydrochloric acid (HCl) to hydrolyze soluble soaps. 2) Chemicals: Carbon tetrachloride (CCl 4 ) reagent grade with an evaporation residue not higher than 5 mg/l (0.005 ) Magnesium sulfate monohydrate. Dry overnight a thin layer of heptahydrate (MgSO 4. 7 H 2 O) in an oven at 150 C. 3) Sample dehydration: Add 25 g of magnesium sulfate monohydrate to the acidified sample in the beaker. Stir to a smooth paste and spread on the sides of beaker. In minutes the paste solidifies. Remove the solids and grind in a mortar. Add the powder to an extraction thimble. Wipe mortar and beaker with filter paper wetted with solvent and add to thimble. 4) Extraction: ml of solvent Boiling stones 15 minutes with the sample immersed in solvent 30 minutes of reflux washing 5) Extract drying: Extraction vessel in an oven at 80 C for 15 minutes. Cool in a desiccator. 6) Weighing: The extract containing vessel with boiling stones is weighed to 0.1 mg proximation. 7) Expression of results: Refer to wet solids, or dry solids if known, as mg/kg. 8) Hydrocarbons: Fatty acids as polar materials can be selectively removed by silica gel (Si0 2 ). Dissolve the residue from oil and grease extraction in 10 ml of carbon tetrachloride and pass through a small glass column (Ø 1 cm 20 cm high) filled with mesh silica gel, only mineral oils are not retained. 9) Reference: APHA-AWWA-WPCF (1985). Standard Methods for the Examination of Water and Wastewater. 16th Ed. Washington D.C., USA /A11 33

38 Extraction of petroleum source rocks During the explorative process in search of petroleum sources special importance assumes the qualitative analysis of petroleum source rocks. In order to evaluate the petroleum potential of a particular drilling site, is carried out an analysis of hydrocarbon compounds contained in soil, clay or in crushed rock samples, coming from petroleum sources. Usually, these tests are also performed on sea bed samples in monitoring marine pollution at various distances from the borehole. The extraction is performed on relatively high sample amounts, because of the low content of extractable matter (usually lower than 1%). Reasons for extraction: -gravimetric and qualitative analysis -sample preparation for GC-MS 1) Sample: Crush the sample and grind it (particles dimension lower than 1 mm). Weigh 15g of sample, with a tolerance margin of +/- 10mg, into an extraction thimble and put a small amount of defatted cotton wadding over the sample. 2) Chemicals: methylene chloride, reagent grade. Cotton: defatted cotton wadding. 3) Extraction: accurately weigh the glass extraction vessel with a tolerance margin of 1 mg. Add 60 ml of methylene chloride. Transfer the thimble with the sample in the extraction unit. Program, for the heating plate, a temperature of 120 C. 40 minutes with thimble immersed in boiling solvent 80 minutes of reflux washing 4) Weighing: dry the vessel containing the extract in an oven at 80 C for 15 minutes. Let cool in a desiccator up to room temperature. Weigh with a tolerance margin of 1 mg. 5) Expression of the results: % extractable matter = W 3 W 2 / W 1 x 100 W 1 = sample weight (g), W 2 = glass extraction vessel weight (tare,g), W 3 = glass extraction vessel weight +residue weight (g) /A11 34

39 Extraction of plasticizer mixtures from vinyl chloride plastics The method is devised for the extraction of monomeric and polymeric plasticizers containing simple phtalate esters from polyvinyl chloride compositions for following gas chromatographic analysis. 1) Sample: 1 to 5 g of polymer grinded to 0.02 mm size (20 US mesh) or 5 by 5 mm squares cut from 0.6 to 2 mm thick sheets. 2) Chemicals: 2+1 mixture by volume of carbon tetrachloride (CCl4) and methanol (MetOH) both reagent grade. Boiling chips 3) Extraction: The sample weighed to 0.1 mg proximation is placed in an extraction thimble and extracted 30 minutes immersed in boiling solvent 30 minutes under reflux washing 4) Extract drying: Extraction vessel with boiling stones in an oven at 80 C for 30 minutes 5) Weighing: The extract containing vessel with dried extract and boiling stones is weighed to 0.1 mg proximation 6) Gas chromatography: For following gas chromatographic analysis the extract is dissolved with chloroform 7) Reference: The American Society for Testing Materials (1975). Extraction and Analysis of Plasticizer Mixtures from Vinyl Chloride Plastics. ASTM Standard Practices. Philadelphia, USA /A11 35

40 Extraction of rosin from paper and paperboard The extracted matter is really colophony but in paper manufacture the term rosin is commonly used. 1) Sample: 5-7 g weighted to 0.01 g proximation, cut into 6x40 mm strips. Heavy paperboard can be cut into thinner layers to improve extraction 2) Sample pretreatment:: If the paper contains carbonates the sample must be immersed 5 minutes into 1 N hydrochloric acid (HCl), washed with water and dried at room temperature. 3) Chemicals: Acidified ethanol, 4 ml of concentrated hydrochloric acid (HCl) into 1 l of 95% ethanol (EtOH) Boiling stones 4) Extraction: ml of solvent Boiling stones 15 minutes with the sample immersed in solvent 30 minutes of reflux washing. 5) Extract drying: Extraction vessel in an oven at 105 C for minutes 6) Weighing: The extraction vessel with the dry extract and boiling stones is weighed to 0.01 g proximation 7) Calculation: % rosin = extract and vessel weight - vessel sample weight weight x 100 8) Reference: TAPPI Standard T Rosin in Paper and Peperboard TAPPI Standards and Suggested Methods. Technical Association of the Pulp and Paper Industry. Atlanta, Georgia, USA /A11 36