**5. Fourier transform infrared spectrum and 2DIR correlation spectroscopy for natural product analysis**

Analysis of natural product with FTIR and 2DIR have to confront with the obstacles coming from the natural texture of the subjects, since their contents consist of water in living cells found leaf, fruit, stem, flower, seed or fruiting body, stalk and sclerotium from medicinal mushroom. The spectroscopy analysis of natural product needs to avoid the interrupting H2 O background and preserve the nature of the features. Therefore, the analysis must apply the correct method chosen as well as appropriate spectrum interpretation on natural product.

There are three choices of FTIR spectrum scanning for natural product. Firstly, mixture with KBr if the natural product is completely dried and without the need for further drying process, such as seed, some rhizome or hash portion of fruiting body. Secondly, ATR is the best choice if the sample is sensitive to the environment conditions when they are being removed or still contain water and mucous secretions. Even though there is a slight refection on the sample surface through the diamond platform, the spectrum created is still sufficient to represent the actual profile when using air as a background. In comparison, the juice of the natural product containing water can be analysed using liquid sample cell with calcium fluoride window. The third method is the FTIR imaging attached to a microscope with a spectroscopy system. The combination enriches the scope of analysis and is more powerful and user-friendly.

No doubt that human error is unavoidable when dealing with natural product analysis. The content(s) may have been lost during the sample processing, poor quality of extraction, physical deterioration and extended period of storage. The more the natural content of the subject model is preserved, the more accurate it can be analysed. These kinds of natural product are found in Malay, Chinese [6] and Ayurvedic medicine such as medicinal mushroom. Many remedies in these communities contain ready to use formulae of the natural products for therapy, e.g. use of turmeric product [7] for blood clotting. FTIR is one of the methods to study the key chemical contents in such treatment.

### **5.1. Analysis of natural product and raw material**

### *5.1.1. Appropriate for quantification of chemical marker analysis*

In order to determine the exact quantity of the chemical(s) involved in the treatment, the Spectrum Quant is especially effective for this purpose, using different concentrations of standards that will generate the range of spectra for the targeted compound. The other algorithms such as PCR+, PLS and Quant C can also be chosen depending on the respective objectives of the study. The results of analysing an unknown sample can be used to predict its concentration by comparing with the range of standards.

### *5.1.2. Exposure to various factors*

The original natural product is the best type of sample for chemical fingerprint profiling, maturity or storage period determination. Time factors such as oxidation, exposure to air, etc., could affect the whole experiment. The sample of natural product analysed using FTIR and 2DIR is prevented from the detrimental effects of these factors.

### *5.1.3. Determination of different geographical origin of natural product using principal component analysis or SIMCA*

**5. Fourier transform infrared spectrum and 2DIR correlation spectroscopy** 

Analysis of natural product with FTIR and 2DIR have to confront with the obstacles coming from the natural texture of the subjects, since their contents consist of water in living cells found leaf, fruit, stem, flower, seed or fruiting body, stalk and sclerotium from medicinal mushroom. The spectroscopy analysis of natural product needs to avoid the interrupting

O background and preserve the nature of the features. Therefore, the analysis must apply the correct method chosen as well as appropriate spectrum interpretation on natural product. There are three choices of FTIR spectrum scanning for natural product. Firstly, mixture with KBr if the natural product is completely dried and without the need for further drying process, such as seed, some rhizome or hash portion of fruiting body. Secondly, ATR is the best choice if the sample is sensitive to the environment conditions when they are being removed or still contain water and mucous secretions. Even though there is a slight refection on the sample surface through the diamond platform, the spectrum created is still sufficient to represent the actual profile when using air as a background. In comparison, the juice of the natural product containing water can be analysed using liquid sample cell with calcium fluoride window. The third method is the FTIR imaging attached to a microscope with a spectroscopy system. The

combination enriches the scope of analysis and is more powerful and user-friendly.

No doubt that human error is unavoidable when dealing with natural product analysis. The content(s) may have been lost during the sample processing, poor quality of extraction, physical deterioration and extended period of storage. The more the natural content of the subject model is preserved, the more accurate it can be analysed. These kinds of natural product are found in Malay, Chinese [6] and Ayurvedic medicine such as medicinal mushroom. Many remedies in these communities contain ready to use formulae of the natural products for therapy, e.g. use of turmeric product [7] for blood clotting. FTIR is one of the methods to study

In order to determine the exact quantity of the chemical(s) involved in the treatment, the Spectrum Quant is especially effective for this purpose, using different concentrations of standards that will generate the range of spectra for the targeted compound. The other algorithms such as PCR+, PLS and Quant C can also be chosen depending on the respective objectives of the study. The results of analysing an unknown sample can be used to predict its concentra-

The original natural product is the best type of sample for chemical fingerprint profiling, maturity or storage period determination. Time factors such as oxidation, exposure to air, etc., could affect the whole experiment. The sample of natural product analysed using FTIR and

**for natural product analysis**

186 Fourier Transforms - High-tech Application and Current Trends

the key chemical contents in such treatment.

**5.1. Analysis of natural product and raw material**

tion by comparing with the range of standards.

*5.1.2. Exposure to various factors*

*5.1.1. Appropriate for quantification of chemical marker analysis*

2DIR is prevented from the detrimental effects of these factors.

H2

Natural product especially the medicinal plants and mushroom of different geographical origin would have a variety of chemical contents with respect to the quality and the quantity. Mean spectral data from randomly chosen plants from a plantation and those from individual plants can be compared. This kind of statistical value could be plotted using a software such as Assured ID which will provide overall comparison of the similar type of natural product collected from different geographical locations. The differences are due to factors such as different types of soil, water, weather, pH of the soil, water, etc. These factors will influence the distance and the percentage of rejection of SIMCA created by three principle components. **Figure 8** showed the example of SIMCA result of *H. sabdariffa* [8, 9] sample from two different locations in Malaysia.

**Figure 8.** Statistic result checking with SIMCA on *H. sabdariffa* collected from two different locations in Malaysia. Thirty five plants were randomly selected from a plantation in Johor (JHR) and Penang (P.P) where there are separately 340 km away. The fruits from each plant were collected in different net bag. Thirty-five net bags of *H. sabdariffa* fruits from two locations were processed and produced the pulverised dry powder for the ATR spectroscopy method. Two locations of samples spectrum were grouped in the software Assure ID with triplicates. The result showed that there are some factors caused the samples from two locations was successfully discriminated with the inter-material distance 3.06. Two spheres sharp of diagrams created ever combined at a portion that joined them together under Principal Component (PC) 1, 2 and 3. The conclusion is the samples from two different locations are different in term of some factors such as water, weather, air, soil, but they still contain certain factors that they are having the same. The data produced can be used for further prediction for geographical origin of the unknown sample.
