You are a chemist in-charge in evaluating the raw materials used in a

Halal certified hotdog factory in Singapore. One of the major ingredients you
purchased is minced beef. Your boss is suspecting that your supplier is mixing
it with cheaper meat (pork and horsemeat). Proposed an analytical method
that will determined if indeed the minced beef is adulterated with pork.

Methods:

1. Detection of horse and donkey meat using PCR detection

In this method real-time PCR assays were developed specifically for horse
and donkey DNA, which are applicable to the detection of even low levels of
horse or donkey meat in commercial products through the aid of optimized
horse and donkey primer probes. Prior to the PCR experimental stage, DNA
was extracted from samples using a special DNA extraction kit, as the collected
samples were subjected to elution and extraction with certain buffers. For
samples in complex meat matrices, DNA was also extracted through another
special method using a DNA purification system. Afterwards, the prepared
samples were subjected to a real-time polymerase chain reaction, which
includes the addition of a TaqMan Buffer, and the presence of the specific
probes that were prepared beforehand. After the reactions, the yielded results
present signals which respond to the optimized probes. The use of DNA, rather
than protein, was proven to be a better method of analysis as proteins are
denatured during heat and pressure processing, making the detection of
species in a sample more difficult.

1. Detection of pork fat in processed foods using HPLC

Another way of detecting of pork fat in processed foods is through HPLC

which separates the triacylglycerols (TG) of animal fats using refractive index
detection. TG is isolated from the pork fat by column chromatography. HPLC
is a good separation technique and has application to the analysis of animal
and vegetable fats and oils. HPLC-profiling of derivatized TG have been
reported for the detection of pork in beef but major disadvantage of the
method is lengthy and needs tedious sample preparation.

1.

FTIR spectroscopy combined with a chemometric technique is an

emerging reliable analytical method to identify meat types present in
processed foods. In the field of halal analysis, the combination of FTIR
spectroscopy and chemometrics has been used for analysis of lard in the binary
mixture with other animal fats such as analysis of pork fat in meatball products
and meatball broth. The FTIR spectra of evaluated samples were acquired in
the mid infrared region (650-4000 cm1) using an ABB MB3000 FTIR
spectrophotometer. The instrument was equipped with a deuterated triglycine
sulfate (DTGS) detector and KBR as the beam splitter. The spectra were
scanned at a resolution of 4 cm1 with 32 scanning. The FTIR spectra were
processed using FTIR software of Horizon MB version 3.013.1 (ABB, Canada).
Quantitative analysis of fat extracted from processed food containing pork
meat was performed using Partial Least Square (PLS) calibration. The major
advantage of this method is that FTIR is rapid, non-destructive and does not
involve laborious sample preparation.