Evaluation of APHA and AOAC II methods for phosphatase in butter and differentiation of milk and microbial phosphatases by agarose-gel electrophoresis

Salted and unsalted butters with 3 levels of phosphatase were prepared with both raw and pasteurized cream containing 36% fat. Test samples were analyzed for phosphatase by the modified method of the American Public Health Association (APHA) and the official AOAC method, 16.256 (1984, 14th Ed., 1990 15th Ed., 946.02). In the APHA method, weighing of solid frozen butter for testing yielded repeatable results. Addition of 0.0-1.0 mg magnesium to the butter had little effect on phosphatase activity in the APHA modified rapid colorimetric method (MRCM), but caused the phosphatase activity to decrease in the AOAC method. Phosphatase in salted and unsalted butters was quite stable at -17 +/- 1 degrees C and at 3.0 +/- 0.5 degrees C; however, within 2 to 4 days, freshly prepared butters stored at 22 +/- 1 degrees C developed reactivated and/or microbial phosphatases that were both heat-labile and heat-stable. At 22 +/- 1 degrees C, frozen butters showed decreased milk phosphatase activity before producing microbial phosphatase. Heat-labile phosphatases in salted and unsalted butters were inactivated at 62.8 degrees C for 10 min, and the phosphatase lability was partially due to the heat-denaturing effect of NaCl in salted butter. Some heat-stable phosphatases in unsalted butter survived at 66 degrees C for 30 min. Differentiation of milk phosphatase from microbial phosphatases was difficult by both methods; however, they were successfully differentiated by the agarose-gel electrophoretic technique.     

Comparison of chemiluminescent and AOAC methods for determining nitrite in commercial cured meat products

A chemiluminescent detector was used to measure nitrite in a variety of commercial cured meat products, using the AOAC sample preparation procedure, 24.041. A comparison of the NaNO2 values obtained by using sulfanilamide/N-(1-naphthyl)ethylenediamine (SAN/NED) reagent, sulfanilamide/1-naphthylamine reagent, and chemiluminescent detection (CLD) revealed no significant differences between the latter 2 detection methods. The AOAC SAN/NED reagent combination gave an average of 24.6% lower NaNO2 results than CLD. Examination of the sample preparation extraction and heating steps indicated that the procedure could not be made more rapid because of the need to destroy residual reductants and release "bound-complexed-reacted" nitrite from the meat samples.     

Evaluation of laboratory performance of the AOAC method for PSP toxin in shellfish

Laboratory performance of the official AOAC method for paralytic shellfish poison (PSP) toxin in shellfish was evaluated. Two series of naturally toxic shellfish split samples were distributed (15 in 1979 and 19 in 1982) to state shellfish-monitoring laboratories which participate in the National Shellfish Sanitation Program. The laboratories performed bioassays on duplicate 100 g portions of each 220 g split sample. Bioassays were consistent among the laboratories and compared favorably with those of previous studies.     

Rapid screening method for alkaline phosphatase activity in cheese: collaborative study

The method developed for developed for determining alkaline phosphatase activity in cheese, in which phenolphthalein monophosphate is used as the substrate, was collaboratively studied. A 7.5% butanol extract of cheese is reacted with phenolphthalein monophosphate; phenolphthalein is released and yields a red solution that is compared visually with a standard (s) prepared from the same extract. Seven collaborators analyzed 8 samples of cheese, in duplicate, by the screening method and Scharer I method. Of the 208 observations returned, only 4 were incorrect. The alkaline phosphatase method has been adopted as official first action.     

Comparison of APHA and elevated temperature enrichment methods for recovery of Vibrio cholerae from oysters: collaborative study

Two methods (the American Public Health Association (APHA) method and the elevated temperature method) for recovery of Vibrio cholerae from oysters were compared in a collaborative study. Oysters were inoculated with high (about 150 cells/g) and low (about 20 cells/g) levels of V. cholerae. The elevated temperature method gave a significantly higher (P less than 0.05) recovery rate and showed greater specificity (P less than 0.01), as determined by the confirmation rate of suspect colonies. The elevated temperature method required only 25% of the labor and materials necessary for the APHA method. The elevated temperature method has been adopted official first action.     

Comparison of paired-ion liquid chromatographic method with AOAC fluorometric and microbiological methods for riboflavin determination in selected foods

A paired-ion liquid chromatographic (LC) technique coupled with fluorometric detection to determine riboflavin in various food matrices is described. Chromatograms of many foods showed 2 peaks of interest due to presence of riboflavin and flavin mononucleotide (FMN). Relatively high levels of FMN were found in raw beef, corned beef, chicken liver, and canned mushrooms. When riboflavin and FMN contents were summed, LC values were comparable to those obtained by the AOAC standard procedures. The LC technique was sensitive, rapid, and simple, yielding a mean standard deviation of 3.1% which was comparable to the AOAC fluorometric method (3.0%) and better than the AOAC microbiological assay (9.6%). Mean spike recoveries were 91.8% for LC compared to 90.5% and 89.6% for the AOAC fluorometric and microbiological methods, respectively.     

凝乳方式对奶油干酪品质的影响

凝乳是干酪加工的关键环节,对奶油干酪的品质具有重要影响。通过对奶油干酪理化成分、产率、涂抹性的测定,比较了酶凝、酸凝、酸-酶互作凝乳3种方式对奶油干酪涂抹性的影响。结果表明,随着酸化程度的加深,奶油干酪的脂肪含量和蛋白质含量逐渐减少,含水率显著增加,校正产率、脂肪回收率和酪蛋白回收率降低。有酶高酸组(RHA)涂抹性最佳,其剪切功、屈服应力都分别为最小(19.23N·s、195.67Pa),与传统酸凝型奶油干酪(NR)相比,RHA的剪切功和屈服应力分别降低了17.04%和27.88%,干酪涂抹性得到明显改善。这为涂抹型奶油干酪的研发提供了技术基础。     

Rapid methods for differentiating reactivated from residual phosphatase in milk and cream: collaborative study.

Three methods for differentiating reactivated from residual phosphatase in milk and cream were collaboratively tested using both magnesium acetate and magnesium chloride for reactivating phosphatase. The methods evaluated were the modified Scharer rapid test, the rapid colorimetric test, and the Rutgers method. Nine collaborators tested 6 unknown milk samples containing reactivated and/or residual phosphatase, and 16 collaborators tested 6 unknown cream samples containing reactivated and/or residual phosphatase. Results indicated that use of magnesium acetate in place of magnesium chloride for reactivating phosphatase improved test results. Visual tests (modified Scharer rapid and Rutgers) predicted correct results when the samples contained high levels of reactivated or residual phosphatase. In borderline cases where the reactivated phosphatase contents of the undiluted control sample and the diluted sample containing Mg were very close, the test results of the visual methods were significantly different from 100% correct results at the alpha=0.05 level. Use of a photoelectric colorimeter or its equivalent for measuring the absorbance in conjunction with the modified Scharer rapid test improved results considerably. The modified Scharer rapid test was adopted official first action.     

Spectrometric and liquid chromatographic determination of natamycin in cheese and cheese rind

Methods for determining natamycin content of cheese rind and cheese are presented. Cheese and rind samples are extracted with methanol and the fat precipitated by cooling the sample solution in methanol-water at -15 to -20 degrees C for ca 1 h. Natamycin levels are measured by UV spectrometric detection at absorbance minimum 311 nm, maximum 317 nm, and at exactly 329 nm, or by LC separation over Lichrosorb RP-8 column with detection at 303 nm. For measuring low levels, a concentration step is provided. The method is applicable to natamycin in cheese rind and in the interior of the cheese. Detection limit is 0.5 mg/kg. The method is suitable for controlling a maximum tolerance of natamycin on the cheese rind, at a level of 1 mg/dm2, and for detecting migration of natamycin into the cheese.     

Recommendations for preparing test samples for AOAC collaborative studies of microbiological procedures for foods

Preparation of test samples for microbial collaborative studies poses problems not encountered in studies on chemical analytes. For Associate Referees who are considering a collaborative study of a microbiological procedure for food analysis, these problems have not been adequately addressed. Types of contamination (natural or artificial), number of test samples required, analyte selection, proper controls, and container selection are addressed herein. The discussion is a supplement to the guidelines contained in the Handbook for AOAC Members.     

Evaluation of a method for assaying sulfonamide antimicrobial residues in cheese: hot-water extraction and liquid chromatography-tandem mass spectrometry

Several sulfonamide antimicrobials (SAAs) are largely used in veterinary medicine. A rapid, specific, and sensitive procedure for determining 12 SAAs in cheese is presented. The method is based on the matrix solid-phase dispersion technique followed by liquid chromatography (LC)-tandem mass spectrometry (MS) equipped with an electrospray ion source. Target compounds were extracted from Mozzarella, Asiago, Parmigiano, Emmenthal, and Camembert cheese samples by 6 mL of water modified with 10% methanol and heated at 120 degrees C. The addition of methanol to hot water served to improve remarkably extraction yields of the most lipophilic SAAs, that is, sulfadimethoxine and sulfaquinoxaline. After acidification and filtration, 100 microL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the multireaction monitoring mode, selecting two precursor-to-product ion transitions for each target compound. Methanol-modified hot water appeared to be an efficient extractant, because absolute recovery ranged between 67 and 88%. Using sulfamoxole as surrogate analyte, recovery of the 12 analytes spiked in the five types of cheese considered at the 50 ng/g level ranged between 75 and 105% with RSD not higher than 11%. Statistical analysis of the mean recovery data showed that the extraction efficiency was not affected by the type of cheese analyzed. This result indicates this method could be applied to other cheese types not considered here. The accuracy of the method was determined at three spike levels, that is, 20, 50, and 100 ng/g, and varied between 73 and 102% with relative standard deviations ranging between 4 and 12%. On the basis of a signal-to-noise ratio of 10, limits of quantification were estimated to be <1 ng/g.     

山东省粮食作物施肥状况的评价

对山东省不同地区1046个农户1997年施肥状况调查表明,山东省粮食作物化肥施用中存在许多不合理之处,农民偏重小麦施肥,小麦化肥氮全省平均280.0kg／hm2,有机肥氮90．4kg／hm2;玉米化肥氮208．5kg／hm2,有机肥氮33.0kg／hm2,均高于合理施氮量.计算结果表明,仅小麦超量施用化肥氮就达40万吨．合理施肥量范围内的农户比例不足20％,70％以上农户超量施肥．小麦玉米轮作周期施氮量已达到吨粮田的施氮水平,但产量只有吨粮田的70％山东省今后提高化肥增产效应需要重点解决这些问题．