This document provides details on the Babcock method for determining the fat content of raw milk. It describes the key equipment needed like Babcock bottles, pipettes, centrifuges, and dividers. It specifies parameters for the bottles like dimensions, graduations, and errors. It also gives procedures for testing the accuracy of bottles and pipettes, adding sulfuric acid to milk in the bottles, and centrifuging to separate the fat for quantification. Parameters are provided for centrifugation speed based on the centrifuge diameter.
This document provides details on the Babcock method for determining the fat content of raw milk. It describes the key equipment needed like Babcock bottles, pipettes, centrifuges, and dividers. It specifies parameters for the bottles like dimensions, graduations, and errors. It also gives procedures for testing the accuracy of bottles and pipettes, adding sulfuric acid to milk in the bottles, and centrifuging to separate the fat for quantification. Parameters are provided for centrifugation speed based on the centrifuge diameter.
This document provides details on the Babcock method for determining the fat content of raw milk. It describes the key equipment needed like Babcock bottles, pipettes, centrifuges, and dividers. It specifies parameters for the bottles like dimensions, graduations, and errors. It also gives procedures for testing the accuracy of bottles and pipettes, adding sulfuric acid to milk in the bottles, and centrifuging to separate the fat for quantification. Parameters are provided for centrifugation speed based on the centrifuge diameter.
This document provides details on the Babcock method for determining the fat content of raw milk. It describes the key equipment needed like Babcock bottles, pipettes, centrifuges, and dividers. It specifies parameters for the bottles like dimensions, graduations, and errors. It also gives procedures for testing the accuracy of bottles and pipettes, adding sulfuric acid to milk in the bottles, and centrifuging to separate the fat for quantification. Parameters are provided for centrifugation speed based on the centrifuge diameter.
AOAC Official Method 989.04 diameter of wheel to achieve 164 ´ g Fat in Raw Milk Diameter of wheel, cm Diameter of wheel, in. rpm Babcock Method 35.6 14 909 Final Action 1920 Revised First Action 1989 40.6 16 848 Revised First Action 1997 45.7 18 800 Final Action 2000 50.8 20 759 (Applicable to raw, whole milk.) 55.9 22 724 Results of the interlaboratory study supporting acceptance of the 61.0 24 693 method (for sections A–D): sr = 0.037; sR = 0.047; RSDr = 0.901%; RSDR = 1.147%; r = 0.105; R = 0.133 A. Principle (3) Testing.—Accuracy of each bottle shall be determined before Known weight of milk is placed into Babcock bottle and H2SO4 is use (usually by manufacturer or certified laboratory). Bottle added. Generated heat releases fat, which is then isolated by calibration accuracy is determined by placing bottle upside down on centrifuging and addition of H2O. Fat is quantitated in graduated Babcock bottle calibration apparatus (Nafis tester or modified Nafis portion of Babcock bottle. Result is expressed as % fat by weight. tester) that is capable of delivering known volumes of Hg into Babcock bottle neck. Bottle calibration apparatus delivery is B. Reagents cal ibrated and vol ume of Hg contained be tween 8 and 4% (a) Sulfuric acid.—Specific gravity 1.82–1.83 at 20°C. (0.800 mL), 4 and 0% (0.800 mL), and 8 and 0% (1.600 mL) marks (b) Mercury.—Purity ³99.99%. Used for testing accuracy of is de ter mined. Ac cu racy of bot tle neck cal i bra tion may be Babcock bottles. determined before or after attachment to bulb. Accuracy should be at C. Apparatus least ±0.04% on scale of 18 g milk-test bottle. [Reference: J. AOAC (a) Standard Babcock milk-test bottle.—8%, 18 g milk-test Int. 78, 463(1995).] bottle, total height 160–170 mm (6.3–6.7 in.). Bottom of bottle is (b) Pipet.—Stan dard milk pipet con forms to fol low ing flat, and axis of neck is vertical when bottle stands on level surface. spec i fi ca tions: (1) Bulb.—Capacity of bulb to junction with neck is ³45 mL. Bulb may be either cylindrical or conical. If cylindrical, od of base Total length . . . . . . . . . . . . . . . . . . . . . . . £350 mm must be 34–36 mm; if conical, od of base must be 31–33 mm, and OD of suction tube . . . . . . . . . . . . . . . . . . . . 6–8 mm maximum diameter, 35–37 mm. Length of suction tube. . . . . . . . . . . . . . . . . . . 130 mm (2) Neck.—Cylindrical and of uniform diameter from ³5 mm OD of delivery tube [must fit into bottle, (a)] . . . . . 4.5–5.0 mm below lowest graduation mark to ³5 mm above highest. Top of neck Length of delivery tube is flared to diameter of ³10 mm. Graduated portion of neck is (slightly longer than bottle neck, must not touch ³75 mm long and is graduated in whole %, 0.5%, and 0.1% from 0.0 milk test portion when delivered) . . . . . . . . . 105–125 mm to 8.0%. Graduations may be etched, with black or dark pigment Distance of graduation mark above bulb . . . . . . . . 15–45 mm annealed to graduation, or may be unetched black or dark lines Nozzle parallel with axis of pipet, but slightly constricted so as to discharge in 5–8 s when filled permanently annealed to glass. Graduation line widths £0.2 mm. with H2O Tenth % graduations are ³3 mm long; 0.5% graduations are ³4 mm Graduation, marked to contain 17.6 mL H2O at 20°C long and project 1 mm to left; and whole % graduations extend at when bottom of meniscus coincides with mark on least half way around neck to right and project ³2 mm to left of 0.1% suction tube graduations. Each whole % is numbered, with numeral placed to left Maximum error in graduation . . . . . . . . . . . . . . £0.05 mL of scale. Vertical line must be etched and annealed with black or dark To test pipet, place tip of pipet against firm rubber surface, clamp pigment or must be etched into dark stripe permanently annealed to pipet in vertical position, and use buret (Class A—graduations glass. Etched vertical line is located 1 mm to right of 0.1% £0.05 mL) to fill pipet to graduation mark with H2O (at 20°C). graduation marks and extends ³1 mm above 8% line and ³1 mm (c) Acid measure.—Device used to measure H2SO4 should deliver below 0% line. If vertical line is etched into dark vertical stripe, then in range from 10 to 20 mL. Use device that can be set to consistently dark vertical stripe must extend ³1 mm above the 8% line and deliver appropriate amount of acid to obtain desired milk acid ³1 mm below 0% line. Dark vertical stripe should be £2.0 mm wide reaction temperature in D. and etched line must be £0.2 mm wide. (d) Centrifuge or “tester.”—Standard centrifuge, however Zero line (£0.2 mm wide) must be etched and annealed with black driven, must be constructed throughout and so mounted as to be or dark pigment, or not darkened but etched into dark horizontal capable, when filled to capacity, of rotating at necessary speed with stripe (£2.0 mm wide) permanently annealed to glass. Zero and minimum vibration and without liability of causing injury or vertical etched lines, with or without annealed pigment, should be of sufficient depth to hold point of dividers. accident. Centrifuge must be heated, electric or otherwise, to Capacity of neck for each whole % on scale is 0.200 mL. 46°–51°C during centrifuging. It must be provided with speed Maximum error of total graduation or any part thereof must not indicator, permanently attached, if possible. Proper rate of rotation exceed 0.008 mL. may be determined by reference to Table 989.04. Rotation speed
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with full centrifuge should be checked periodically with tachometer. (c) Testing milk test portions.—To test bottle from (a) containing “Diameter of wheel” means distance between inside bottoms of tempered milk, add appropriate amount of H2SO4 [as determined in opposite cups measured through center of rotation of centrifuge (b)] by delivering acid in one delivery that washes all traces of milk wheel while cups are horizontally extended. Relative centrifugal into bulb and cleanly layers acid under milk. Immediately shake bottle by hand rotation as in (b), until all traces of curd disappear. force is approximately 164 ´ g. Place bottle in Babcock bottle shaker set at medium speed. Continue (e) Dividers or calipers.—For measuring fat column. to add acid to all test portions, and then shake full set 1 additional (f) Water bath for test bottles.—Provided with thermometer and min. Temperature of milk plus acid in first bottle should not be less device to maintain temperature of fat column at 48° ± 1°C. than 50.5°C at time bottles are transferred to centrifuge. (g) Wa ter bath for tem per ing milk sam ples prior to Place bot tles in heated cen tri fuge, coun ter-balance, and weighing.—Provided with thermometer and device to maintain centrifuge 5 min after proper speed is reached. Add distilled water at temperature of milk at 38° ± 1°C. 50°–52°C until bulb of bottle is filled. Centrifuge 2 min. Add (h) Supply of hot water added after 1st and 2nd distilled water at 50°–52°C until top of fat column approaches 7% centrifugations.—Provided with thermometer, device to maintain mark on bottle neck. Centrifuge 1 min longer at ca 51°C. Transfer temperature of distilled H2O at 50°–52°C, and device to deliver H2O bottle to water bath maintained at 48° ± 1°C, immerse bottle to level into Babcock bottles. slightly above top of fat column, and leave in water bath until (i) Bottle shaker.—Variable speed and matched to maximum column is at equilibrium and lower fat surface assumes final convex capacity of centrifuge. form (³5 min). (j) Digital thermometer for measurement of milk-acid reaction Remove 1 bottle from bath, wipe dry, and with aid of reading light temperature.—Capable of reading temperature to the nearest degree and magnification, C(k), use dividers or calipers to quickly measure in range of 100°–120°C. Use acid-resistant probe, with small fat column (before it begins to cool and contract). Place caliper diameter (£0.5 mm) to ensure rapid response time. Length of points in vertical line on neck of bottle, with 1 point at lowest surface temperature probe should be such that its tip is ca 1 cm above bottom of lower meniscus and other point at top of upper meniscus. Without of bottle, when fully inserted. changing distance between 2 points on calipers, move calipers down (k) Reading light with magnifying glass.—As background when bottle neck until lower point rests in etched horizontal graduation measuring fat columns. Light should be diffused (soft white color) mark at 0%. Place upper point of calipers against bottle graduation and provide illumination from angles above and below level of fat and read test in % to nearest 0.05%. Repeat for each bottle. column. Magnification device must be used to aid reading. Fat column, at time of measurement, should be translucent, (l) Tachometer.—For determining rotation speed of centrifuge golden-yellow or amber, and free of visible suspended particles. (capable of measuring between 600–1000 rpm). Reject all tests in which fat column is milky or shows presence of D. Determination curd or charred matter, or in which meniscus is indistinct or (a) Test sample preparation and temperature adjustment.—Place distorted; repeat test, adjusting volume of H2SO4 added, to obtain test sample in water bath at 38° ± 1°C. Level of H2O should be at or proper color and milk-acid reaction temperature. above milk level. Mix test sample 10´ by inversion. If fat line References: JAOAC 8, 4(1924); 8, 471(1925); 56, 1401(1973); remains on inside surface of container, run hot tap water (ca 58, 949(1975); 71, 898(1988). 50°–60°C) over outside surface for 15–20 s. Mix test sample J. AOAC Int. 80, 845(1997). thoroughly by inversion and pipet immediately. Do not allow test E. Alternative Apparatus (First Action 1986; Final Action 1991) samples to remain in water bath more than 15 min after reaching 38°C. (a) Gercock-style milkfat test bottle.—(Available from Forcoven With pipet, C(b), transfer 17.6 ± 0.05 mL prepared test portion at Products, Inc., PO Box 1556, Humble, TX 77338, USA.) 5%, 38°C to milk-test bottle. Also, transfer 17.6 ± 0.05 mL milk at 38°C 162 ± 3 mm high. Bottom of bottle is flat, and axis of flat graduated to another milk test bottle for measurement of milk-acid reaction tube is vertical when bottle stands on level surface. Quantity of milk temperature. Blow out milk in pipet tip ca 30 s after free outflow charge is 18 g; 18 g is acid-etched and pigmented in clear space above 5% graduated line. Specifications: ceases. Adjust temperature of milk in bottle to 20°–22°C. Adjust (1) Bulb.—Capacity of bulb to junction with neck must be temperature of H2SO4 to 20°–22°C. ³45 mL. Shape of bulb is cylindrical, 36 ± 0.5 mm od. Wall thickness (b) Mea sure ment of milk-acid re ac tion tem per a ture and determination of amount of H2SO4 to use.—Before testing group of 1.4 ± 0.2 mm. Word “Sealed”, manufacturer’s number or name, and test portions, determine correct amount of acid to be used in analysis matted marking spot must be legibly and permanently marked on by measuring milk-acid reaction temperature. Add 17.5 mL H2SO4 surface. to bottle from (a) containing tempered milk test portion prepared for (2) Side filling tube.—8 mm od, 0.1 mm wall thickness. To enter measurement of milk-acid reaction temperature. Add acid in one bulb below junction of bulb and neck of flat graduated tube along delivery that washes all traces of milk into bulb and cleanly layers curved portion of bulb and with minimum angle extend to center of acid under milk. Fully insert digital thermometer probe down bottle bulb and maintain vertical axis for 40 mm to height of 3 ± 2 mm neck and immediately shake by hand rotation until all traces of curd above bottom of bottle. disappear. Peak reaction temperature should be 108° ± 2°C. Adjust (3) Neck.—Graduated portion of flat tube is uniform cross amount of H2SO4 added, until reaction temperature is within this section ³4 mm beyond extremes of graduated scale. Length of range and fat column is translucent golden-yellow to amber. graduated portion of flat neck ³75 mm. Graduated at whole %, 0.5, Amount of acid required may vary between technicians and between 0.1, and 0.05%. Graduations are acid-etched, with black pigment batches of acid. annealed to graduation. Graduation widths ³0.2 mm. Each whole %
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and 0.5% line is numbered on left above line and these lines extend Length of nozzle . . . . . . . . . . . . . . . . . . . . 15–20 mm across flat surface. Smallest graduated line is ³2.5 mm. Intermediate Delivery time . . . . . . . . . . . . . . . . . . . . . . . . 5–8 s graduation line extends 1 ± 0.2 mm equally on both sides of smallest graduated line. Graduation: For raw milk. . . . . . . . . . . . . To deliver 17.6 mL at 20°C (4) Vertical line.—Vertical line is acid-etched and pigmented on when bottom of meniscus coincides right side of graduated flat tube. This line touches intermediate lines with mark on suction tube at their extreme right end and extends beyond 0 and 0.5% lines For processed milk . . . . . . . . . To deliver 17.6 mL at 38°C ³3 mm. Width of vertical lines £0.2 mm. when bottom of meniscus coincides (5) Error.—Maximum error of total graduation or any part with mark on suction tube thereof must not exceed volume of smallest unit of graduation (= Maximum error . . . . . . . . . . . . . . . . . . . . £ 0.05 mL 0.01 mL). Permanent markings on bulb: (b) Gercock-style pipet for raw and processed milk.—To conform For raw milk . . . . . . . . . . . . . . . . . . . . . “Sealed,” to following specifications: manufacturer’s No. or name, TD 17.6 mL at 20°C, Total length: tolerance 0.05 mL, 5–8 s For raw milk . . . . . . . . . . . . . . . . . . . . . £280 mm For processed milk . . . . . . . . . . . . . . . . . . . “Sealed,” For processed milk . . . . . . . . . . . . . . . . . . ³280 mm manufacturer’s No. or name, TD 17.6 mL at 38°C, Suction tube diameter . . . . . . . . . . . . . . . . 7 ± 0.2 mm od tolerance 0.05 mL, 5–8 s Wall thickness . . . . . . . . . . . . . . . . . . . 0.9 ± 0.04 mm Suction tube length . . . . . . . . . . . . . . . . . . . . 130 mm Reference: JAOAC 69, 831(1986). Delivery tube diameter . . . . . . . . . . . . . . . 5 ± 0.2 mm od Distance of graduation mark above bulb . . . . . . . . 15–45 mm Revised: March 2002