The proteases in cow’s milk have a great influence on the flavor and quality, especially on the shelf life of dairy products, but there is no uniform reference standard for the detection of protease activity in dairy products. This paper elaborates the sources, types and characteristics of protease in cow’s milk, and summarizes recent progress in the detection of the activity of different types of protease in cow’s milk, pointing out that the precise detection of protease activity in milk has great significance for dairy production and quality assurance.

An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous quantitative determination of fumonisin B1 (FB1), fumonisin B2 (FB2) and fumonisin B3 (FB3) residues in milk powder and milk has been established. The samples were extracted with methanol-acetic acid (98:2, V/V), diluted, cleaned up by immunoaffinity chromatography (IAC), and blown to dryness under nitrogen. The residue was re-dissolved and detected using an electrospray ionization source (EIS) in the positive ion mode with multiple reaction monitoring (MRM) and quantitative analysis was performed by the isotope-labeled internal standard method. The limit of quantification was 10 μg/kg for FB1, FB2 and FB3 in milk powder and milk. The recoveries at spiked concentration levels of 10–400 μg/kg were 92.8%–107.5%, with relative standard deviations (RSD) of 2.0%–4.3%. The matrix effects were 96.25%–122.84%. This method was characterized by short analysis time and was suitable for the determination of fumonisin residues in different dairy products.

Abnormal analysis of trace elements and heavy metal over-standard in dairy products are major concerns in the dairy industry, and how to detect elements of concern in dairy products quickly and accurately has become the focus of attention. Detection of elements in milk and dairy products is a prerequisite to ensure product quality and safety. Therefore, the pretreatment and detection methods specified in the Chinese and international standards for elements in milk and dairy products are reviewed. The principle, pros and cons of each method are elaborated so as to provide a reference for the detection of elements in the dairy industry.

A method for the determination of stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside F, dulcoside A, rubusoside and steviolbioside in fermented milk by high performance liquid chromatography (HPLC) was developed and validated. The chromatographic conditions and sample pretreatment conditions were optimized. Samples were ultrasonically extracted with water for 20 min and for another 10 min after addition of 30% acetonitrile in water bath at 45 ℃. Potassium ferrocyanide and zinc acetate were added to precipitate proteins. The chromatographic separation was achieved on a C18 with gradient elution using a mobile phase consisting of sodium phosphate buffer and acetonitrile, and the analytes were detected by an ultraviolet (UV) detector at 210 nm. The results showed that the method was linear in the concentration range of 1–50 μg/mL, the limit of detection (LOD, RS/N = 3) was 3.0 mg/kg, and the limit of quantitation (LOQ, RS/N = 10) was 10.0 mg/kg. The average recoveries of all analytes spiked at 15–200 mg/kg were between 92.0% and 103.1%, with relative standard deviation (RSDs) (n = 6) less than 3.0%. This method was rapid, accurate, stable, sensitive, and suitable for the analysis of the content of steviol glycosides in fermented milk.

A reversed-phase high performance liquid chromatography (RP-HPLC) method was established for the determination of 6S-5-methyltetrahydrofolate calcium in milk powder. After protein precipitation with trichloroacetic acid solution, samples were extracted with ascorbic acid solution in a hot water bath, and the extract was separated on a C18 reversed-phase column by gradient elution using a mobile phase composed of 0.1% trifluoroacetic acid solution and methanol, and the detection wavelength was set at 280 nm. Quantitative analysis was carried out by the single standard method. The results showed that the limit of quantification and detection of 6S-5-methyltetrahydrofolate calcium in milk powder samples were 150 and 50 μg/100 g, respectively. The recoveries for samples spiked at levels ranging from 150 to 600 μg/100 g were 95.0%–109.1% with relative standard deviations (RSDs) between 1.26% and 4.04%. This method is simple, accurate, with high recovery and reproducibility, and suitable for the determination of 6S-5-methyltetrahydrofolate calcium in milk powder.

The objective of this study was to establish a method for detecting the residue of dicamba in cow milk by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The sample was extracted with 0.5% formic acid in acetonitrile, and the extract was purified by sodium chloride salting out, and separated on an Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm) by gradient elution using a mobile phase composed of acetonitrile and 0.01% formic acid aqueous solution. Identification and quantification were achieved using an electrospray ionization source in the negative ion mode with multiple reaction monitoring (MRM). Under optimized pretreatment and instrumental conditions, good linearity was observed in the concentration range of 5.0–250.0 μg/L with correlation coefficient (r) more than 0.999. The limit of detection (LOD) and limit of quantification (LOQ) of the proposed method were 5 and 10 μg/kg, respectively. The recoveries of dicamba at spiked concentration levels of 10, 20 and 200 μg/kg were 104.3%, 101.5% and 96.2% with relative standard deviations (n = 6) of 3.0%, 2.0% and 1.5%, respectively. This method is simple, accurate, repeatable, and suitable for the rapid detection of dicamba in milk.

Being rich in nutrients, dairy products provide a good environment for the growth of Staphylococcus aureus, Salmonella, Listeria monocytogenes, Cronobacter and other foodborne pathogens, which poses a serious threat to the safety of dairy products. Therefore, rapid and accurate detection of foodborne pathogens in dairy products is essential to the prevention of foodborne pathogens. In recent years, analytical techniques including molecular biology and immunoassays have rapidly developed and attracted much attention due to their advantages of high sensitivity, rapidity and simplicity compared with the traditional food pathogen detection methods, which are cumbersome and time-consuming. In this paper, recent developments and applications of foodborne pathogen detection techniques are reviewed, and future trends were discussed in order to provide a reference for the rapid detection of foodborne pathogens in dairy products.

A reversed-phase high performance liquid chromatographic (HPLC) method was established for the determination of erythrosine in fermented milk. After enzymatic hydrolysis, the sample was extracted with anhydrous ethanol, and purified by solid phase extraction using an HLB column under alkaline conditions. The separation was performed on a C18 reversephase column by gradient elution using a mobile phase comprising 20 mmol/L ammonium acetate buffer solution (pH 6.5) and methanol. The detection wavelength was set at 520 nm, and quantification was performed by the external standard method. The results showed that the linear range for erythrosine was 0.05–20.00 μg/mL. The limit of quantitation (LOQ) and the limit of detection (LOD) of erythrosine in fermented milk samples were 0.2 and 0.1 mg/kg, respectively. The recoveries from fermented milk samples spiked at levels of 0.2–2.0 mg/kg were 96.5%–105.6%, and the relative standard deviations (RSD) were 1.87%–2.21%. The method is simple and accurate, has high recovery and good repeatability, and is suitable for the determination of erythrosine in fermented milk.

The measurement uncertainty was estimated for the determination of 1-oleic-2-palmitic-3-linoleic acid triglyceride (OPL) content in milk powder by gas chromatography (GC). According to the Measurement and Expression of Uncertainty in Measurement (JJF 1059.1—2012) and the Guidance on Quantifying Uncertainty in Chemical Analysis (CNAS-GL 006: 2019), a mathematical model was established to analyze the sources of measurement uncertainty. The major factors affecting the measurement uncertainty were evaluated. Results showed that the average OPL content of six milk powder samples was measured to be 1.172 g/100 g. The relative expanded uncertainty (urel) was 0.023 6 with a coverage probability of approximately 95%. The standard uncertainty arising from sample preparation was the main source of uncertainty.

A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for the determination of nonylphenol in milk powder. Samples were extracted with acetonitrile and cleaned up with a solid phase extraction column. The analytes were separated on a Waters ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm), detected by electrospray ionization in the multiple reaction monitoring (MRM) mode, and quantified by an external standard method. The results showed that good linearity was observed in the concentration range of 1–200 ng/mL (R2 > 0.99). The limit of detection（LOD, RS/N = 3）was 2 μg/kg and the limit of quantitation (LOQ, RS/N = 10）was 5 μg/kg. The recoveries at different spiked levels were between 81.10% and 107.39% with relative standard deviation (RSD) ranging from 1.97% to 9.73%. This method has the advantages of accurate results, small background interference, simple operation, good stability and short pretreatment time, and is useful for the detection of nonylphenol in milk powder.

A small amount of bacterial spores with strong tolerance still exist in raw milk after sterilization and can multiply under certain conditions, causing a serious impact on product quality and consumer health. In this study, raw milk samples containing Bacillus were collected to isolate aerobic Bacillus and thermophilic aerobic Bacillus. The isolates obtained were identified based on their morphological, physiological, biochemical and molecular biological characteristics, and the heat resistance of the isolated thermophilic aerobic Bacillus was studied. Results showed that 10 strains of Bacillus were isolated from the milk samples, of which three were thermophilic aerobic Bacillus and seven were aerobic Bacillus. Among them, Bacillus coagulans and Bacillus clausii could tolerate heat treatment at 115 ℃ for 15 min. All Bacillus in raw milk could be killed by heat treatment at 121 ℃ for 20 min which decreased the number of spores by four orders of magnitude. The dominant Bacillus species in raw milk were Bacillus subtilis, Bacillus cereus, Bacillus pumilus, Bacillus licheniformis, Bacillus circulans, Bacillus gelatini, Bacillus amyloliquefaciens, Bacillus coagulans, and Bacillus clausii. Bacillus coagulans and Bacillus clausii had stronger heat resistance and could tolerate heat treatment at 115 ℃ for 15 min.

A high performance liquid chromatographic (HPLC) method was developed for the determination of naringin in milk and dairy products. Samples were extracted with methanol, and the analytes were separated on a C18 column using a mobile phase consisting of 0.1% acetic acid aqueous solution and acetonitrile (75:25, V/V), detected by a diode array detector at 284 nm and quantified by an external standard method. The results showed that the linearity of this method was good in the concentration range of 5–500 μg/mL with a correlation coefficient of 0.999 9. The recoveries of powdered and liquid milk power and yogurt spiked at 0.02, 0.04, and 0.08 g/100 g were 96.2%–103.2%, 96.3%–103.7%, and 99.7%–104.1%, respectively, with relative standard deviations (n = 6) of 1.12%–2.31%, 1.25%–1.85%, and 1.24%–1.66%, respectively. The reported method is simple, rapid, accurate, repeatable, and suitable for the determination of naringin in milk and dairy products.

Dairy products are favored by more and more consumers because of their rich nutrients and good taste, and have become an important part of consumers’ dietary structure. At the same time, the food safety of dairy products has become the focus of consumers’ attention. Therefore, controlling dairy food safety is a top priority for every dairy company. This article expounds on the food safety control measures for ensuring quality and safety in each link of the dairy product production process. To ensure the safety of milk sources, pasture construction and management and risk control during breeding activities need to be considered. The control of raw and auxiliary materials needs to be carried out by considering the acceptance system, storage of raw and auxiliary materials, and upstream supplier management and control. The control of the production process needs to be carried out from the aspects of personnel, equipment, ingredient and environmental management. In addition, this review highlights the importance of big data technology in dairy food safety risk analysis. We expect that this review will be helpful to relevant enterprises in improving their work on food safety management.

A gas chromatographic (GC) method for the determination of 1-oleic-2-palmitic-3-linoleic-triglyceride (OPL) in milk powder was established. The samples were extracted with ether and petroleum ether, and the extract was concentrated to near dryness, re-dissolved in n-heptane, separated on a CP-TAP CB elastic quartz capillary column, detected by a hydrogen flame ionization detector, and quantified by an external standard method. The column temperature was programmed to rise. The inlet temperature was set at 330 ℃. The samples (1.0 μL) were injected using the split mode (1:10). The detector temperature was set at 360 ℃. Nitrogen was used as the carrier gas. The makeup flow rate (nitrogen) was 25 mL/min. The recovery of OPL from spiked milk powder at 1–6 g/100 g ranged from 95.2% to 101.9%, and the relative standard deviations (RSDs) for precision ranged from 1.289% to 1.998%. The limit of detection (LOD) and limit of quantitation (LOQ) of the proposed method were 0.2 and 1.0 g/100 g, respectively. This method, therefore, is simple, accurate, repeatable, and suitable for the determination of OPL in milk powder.

Sample pretreatment is a key step in element testing, which directly affects the precision and accuracy of analytical results. In order to meet the requirements for the detection of various elements, in this study, we used microwave digestion combined with inductively coupled plasma-mass spectrometry (ICP-MS) operated in the collision reaction mode to determine the content of multiple elements in milk powder, and investigated the influence of sample pretreatment conditions (acid removal time, sample size, and the amount of residual acid) on the detection results. Compared to removing acid mist only, better results were obtained under the following conditions: acid removal temperature was 160 ℃, acid removal time was 97.5 min, and sample size was 0.250 0 g, and the amount of residual acid was 1.0 mL. This pretreatment process is characterized by good stability, high precision and high accuracy.

Inductively coupled plasma-mass spectrometry (ICP-MS) was used to measure the aluminum content in the quality control sample of milk powder, and an X-control chart was established for runaway judgement and handling during the operation process. The precision and median line of the control chart for new data points obtained one year later were evaluated using Minitab software with test for equal variance and analysis of variance. The standard deviation for the previous 25 data points was 0.186 mg/kg compared to 0.126 mg/kg for the 35 new ones, while the median values of the two groups did not differ from each other. More reliable control limit was calculated based on all the data points, and the new median value was 1.334 mg/kg with a standard deviation of 0.154 mg/kg. The decrease in standard deviation suggested that the precision was improved with smaller data fluctuations. In conclusion, calculation of new and more reliable control limit from more data is useful to find obvious systematic errors, abnormal data and slow changing trend in a timely and effective way and can provide evidence to decide whether data should be retained or removed so as to directly reflect the stability and tendency of the detection.

A gas chromatographic (GC) method to determine the contents of 16 trans fatty acids in ice cream was established. The 2.0 g samples were pretreated sequentially with 8 mL of hydrochloric acid and 10 mL of anhydrous ethanol, separated on an SP-2560 capillary column, detected by GC with a flame ionization detector (FID) and quantified by the peak area normalization method. Under optimized chromatographic conditions, the limit of detection and limit of quantification of the proposed method were 0.012% and 0.024%, respectively, and it allowed effective separation of trans fatty acid methyl esters from cis fatty acid methyl esters and saturated fatty acid methyl esters.

Iodine is an important index of the nutritional quality of milk. Milk and dairy products are important dietary sources of iodine for humans, which has always aroused extensive concern. With the continuous exploration of nutrients in foods, research on iodine in milk has been increasingly intensified and refined. This paper reviews the recent literature on the factors that affects the iodine content of milk and the analytical methods that have been applied to determine it, which is expected to provide a reference for researchers in this industry and also provide a theoretical basis for the labelling of iodine in infant formula preparation.

In order to provide a scientific rationale for the quality control in the determination of aerobic plate count (APC) in milk, the measurement uncertainty was estimated. According to the national standard of China (GB 4789.2—2016), APC in samples was detected. The sources of uncertainty were analyzed according to Evaluation and Expression of Uncertainty in Measurement (JJF 1059.1—2012) and a mathematical model for uncertainty evaluation was established. The introduced uncertainty components were evaluated and finally the total combined uncertainty was established by combination of individual components. The results showed that the extended uncertainty for APC quantification in milk samples was 0.043 4. The logarithm of APC was [3.924, 4.010], and from this, APC was calculated to be 8 395–10 233 CFU/mL. In conclusion, this procedure can effectively assess the measurement uncertainty in the determination of APC in milk.

The detection of pesticide residues in dairy products is an important measure to ensure the quality and safety of dairy products. Currently, QuEChERS (quick, easy, cheap, effective, rugged, safe) is widely used as a sample pretreatment for all existing methods specified in the national standards for the detection of pesticide residues in dairy products. In this paper, we review these methods with a special focus on the principle and advantages of QuEChERS as well as its application in the detection of pesticide residues in dairy products.

In order to develop a new method to determine nitrite and nitrate in milk and milk products, some improvements were made on the ion chromatography method specified in the national standard GB 5009.33-2016 Determination of Nitrite and Nitrate in Food. Samples were extracted with ultra-pure water and added with acetic acid to precipitate proteins before being chromatographed on AS11-HC column and detected with a conductivity detector. Under the above conditions, the calibration curves for nitrite and nitrate were linear with correlation coefficients greater than 0.99. The limit of quantitation (LOQ) for nitrite and nitrate was 0.25 and 0.8 mg/kg in milk, and 1 and 5 mg/kg in milk powder, respectively. The recoveries for spiked samples of milk powder and milk were from 80% to 110%. The improved method was simple to operate and cheap with ideal recovery and high precision. It could avoid potential problems encountered in the use of the traditional method and allow for more rapid and accurate detection of low contents of nitrite and nitrate in milk and milk products.

A method for the determination of zeaxanthin in milk powder was developed using high performance liquid chromatography (HPLC). The sample was saponified with potassium hydroxide solution to release zeaxanthin, extracted with a mixed solvent of ethyl ether, n-hexane and cyclohexane and frozen for degreasing. The chromatographic separation was performed on a C30 column with gradient elution. The analyte was detected with an ultraviolet detector and quantified by an external standard method. The limit of quantification was 100 μg/100 g and the limit of detection was 30 μg/100 g for zeaxanthin in milk powder. The method recoveries at spiked concentration levels of 90–811 μg/100 g were 85.8%–102.2%, with relative standard deviations ranging from 1.43% to 2.76%. This method was accurate, repeatable, precise and sensitive.

A spectrophotometric method for the determination of tea polyphenols in milk tea powder was developed by using Folin-phenol reagent. Crucial operating parameters such as extraction reagent, number of extraction cycles, temperature and derivatization reagent were investigated. Extraction using 70% methanol solution as the extraction reagent in a hot water bath at 70 ℃ performed twice and oxidization of the resulting extract with Folin-phenol reagent proved optimal. The detection wavelength was selected as 765 nm, and gallic acid was used as the calibration standard for the quantitation of tea polyphenols. The recoveries for whole milk powder spiked at concentration levels of 850–4 500 mg/kg were in the range of 98.9%–104.9%. Good stability and accuracy were achieved. When milk tea powder samples with different contents of tea polyphenols were detected in six replicates, relative standard deviations (RSDs) of 0.29%–0.84% were obtained, indicating good precision. This method proved to be accurate, easy to operate and repeatable with simple and easily available instruments, and thus suitable for the analysis of tea polyphenols in milk tea powder.

By analyzing some factors involved in the determination of lead in milk powder by inductively coupled plasma mass spectrometry (ICP-MS) such as containers and reagents used, an ultrasonic cleaning method was established to effectively reduce the blank value. The method gave a detection limit of 0.005 mg/kg, which was much lower than that (0.02 mg/kg) of traditional methods. The recoveries of samples spiked at concentration levels of 0.005–0.015 mg/kg ranged from 86.9% to 119.0% with relative standard deviations (RSDs) of 3.1%–12.3%, meeting the requirements for methodological validation. Hence, the method proved to be overall feasible and suitable for the detection of lead in milk powder with reduced detection limit.

In recent years, the addition of probiotics to milk powder has gradually become a research focus. In order to ensure the product quality and safety, a microbiological detection method for Lactobacillus fermentum CECT5716 in milk powder has been established. It was found that the population of Lactobacillus fermentum in milk powder could be accurately counted by spreading serial 10-fold dilutions in buffered peptone water (BPW) onto MRS plates and culturing them under anaerobic conditions at (36 ± 1) ℃ for (48 ± 2) h, with clear discrimination from inoculated bifidobacteria. This method was more suitable for counting Lactobacillus fermentum than the Chinese national standard method (GB 4789.35?2016), and had the advantages of high discrimination capacity, short culture time, simple operation, good repeatability, and high accuracy. This method also included 16S rDNA sequence analysis of the strain for its subsequent molecular identification. There was no significant difference in the accuracy of this method versus the national standard method in detecting 20 samples prepared in our laboratory (P = 0.127).

An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous quantitative determination of 21 β-agonists in milk. The samples were hydrolyzed and cleaned up a solid-phase extraction column. The analytes were separated by UPLC with an Acquity UPLC BEH C18 column (50 mm × 2.1 mm, 1.7 μm) using a gradient mobile phase consisting of 0.1% formic acid aqueous solution and methanol (10 : 90, V/V). Identification and quantification were achieved by UPLC-MS/MS with multiple reaction monitoring (MRM) under the positive ion mode The limit of quantitation of all β-agonists was 0.05 μg/kg. The average recovery varied from 61.0% to 124.3% at spiked concentration levels of 0.05, 0.10 and 0.50 μg/kg with relative standard deviations (RSDs) of 2.39%–10.66%. Our results demonstrate that the method is stable, accurate and sensitive, and can be used for the determination of the 21 β-agonists residues in milk.

A reversed-phase high performance liquid chromatography (RP-HPLC) method for the simultaneous quantitative determination of vanillin, methyl vanillin, ethyl vanillin and coumarin in milk and dairy products has been established. Vanillin compounds from samples were extracted with acetonitrile, blown to dryness under nitrogen, and then cleaned up with n-hexane. The chromatographic separation was achieved using a mixture of 20 mmoL/L ammonium acetate (pH 5.6), acetonitrile and methanol as the mobile phase with gradient elution on a C18 column. The analysis was carried out using a UV detector at a wavelength of 267 nm and the analytes were quantified by an external standard method. Good linear relationships in the range of 0.05–5.00 μg/mL were observed for all analytes. The limit of quantification was 0.2 mg/kg and the limit of detection was 0.06 mg/kg for the four analytes in liquid milk, milk powder, yogurt, lactic acid fermented beverage, ice cream, and cheese. The recoveries for vanillin, methyl vanillin, ethyl vanillin and coumarin in these samples at spiked concentration levels of 0.2–2.0 mg/kg were 80.6%–110.0%, 80.1%–109.6%, 80.4%–109.5% and 80.1%–105.9%, with relative standard deviations of 1.87%–7.70%, 1.45%–9.80%, 1.66%–9.52% and 1.16%–9.52%, respectively. This method was rapid, accurate, repeatable and simple and could be applied to determine vanillin compounds in milk and dairy products.

This paper summarizes and compares the maximum residue limits for pyrethroid pesticides in milk products in China and several other countries. At the same time, the main factors affecting the accuracy of the determination of pyrethroid pesticide residues in dairy products, including calibration curve fitting and experimental repeatability, are determined through calculation of the measurement uncertainty. Furthermore, it can be argued that the most fundamental influencing factor is the sample pretreatment method.

The residues of ochratoxin A, deoxynivalenol, aflatoxins B1 and zearalenone in dairy feeds were simultaneously detected using biochip array technology. The method accuracy, precision and reproducibility were evaluated. The results obtained that the correlation coefficient of the calibration curve for each analyte was greater than 0.99. The recoveries for quality control sample and negative sample at two spiked levels varied from 80% to 120%. The coefficient of variance (CV) was less than 15% for accuracy and reproducibility and was less than 10% for reproducibility. Compared with high performance liquid chromatography, this method gave consistent results with a simpler sample pretreatment procedure. In conclusion, the biochip array method was simple and accurate and it could provide an efficient and reliable tool for mass screening of samples.

Inductively coupled plasma emission spectrometry (ICP-OES) was used to determine the contents of 12 trace elements (K, Na, Ca, Mg, Fe, Zn, Cu, Mn, P, Sr, Al, and Cr) in premixed feed and crude feed for dairy cows. Samples were pretreated with microwave digestion before being analyzed. The calibration curves of all elements were linear with correlation coefficients above 0.999 8. The spiked recoveries for this method were 90.0%–106.9%, and the coefficients of variance (CV) for six replicate determinations were 0.69%–8.70%. With the advantages of low limits of detection and high sensitivity and accuracy, this method could be useful for the simultaneous determination of multiple trace elements in dairy feeds.