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Aplikasi Kemometrika Yang Dikombinasikan Dengan Spektroskopi FTIR Dan HPLC Untuk Evaluasi Aktivitas Antioksidan Dan Antibakteri Rimpang Jahe Emprit (Zingiber officinale var. Amarum)

ANITA AGUSTINA STYAWAN, Prof. Dr. apt. Abdul Rohman, M.Si; Prof. Dr. apt. Ratna Asmah Susidarti, MS; Dr. apt. Purwanto, M.Sc

2024 | Disertasi | S3 Ilmu Farmasi

Jahe emprit atau Zingiber officinale var. Amarum digunakan oleh masyarakat sebagai obat tradisional dengan tujuan terapi. Penggunaan bahan herbal seringkali tidak konsisten efeknya, karena adanya perbedaan lokasi tumbuh yang dapat mempengaruhi kandungan kimia obat. Oleh karena itu, data mengenai profil kandungan kimia menjadi   penting sebagai proses standardisasi. Salah satu upaya dalam hal standardisasi bahan alam dengan analisis profil metabolitnya. Penelitian ini bertujuan untuk mengevaluasi aktivitas antioksidan rimpang jahe emprit, mengklasifikasikan serbuk jahe emprit dari lokasi tumbuh berbeda, mengkorelasikan antara spektroskopi FTIR dengan pendekatan kemometrik, dan mengevaluasi aktivitas anti bakteri dari minyak atsiri jahe emprit.

Serbuk jahe emprit diuji aktivitas penangkapan radikal bebas menggunakan metode 2,2-difenil-1-pikrilhidrazil (DPPH), ditetapkan kadar flavonoid total dan fenolik totalnya. Serbuk jahe emprit juga dilakukan pengukuran spektra FTIR yang dikombinasikan dengan HPLC untuk dikorelasikan dengan kadar gingerol. Profil minyak atsiri jahe emprit dari 14 lokasi tumbuh diperoleh menggunakan Gas Chromathography Mass Spectometry (GC-MS) serta dianalisis dengan kalibrasi multivariat Principal Component Analysis (PCA) dan Orthogonal Partial Least Squares (OPLS) menggunakan software SIMCA. Aktivitas antibakteri minyak atsiri jahe emprit dilakukan dengan metode mikrodilusi pada bakteri Staphylococcus aureus (ATCC 25923) dan Escherichia coli (ATCC 25922). Analisis aktivitas antibakteri ditentukan dengan metode analisis probit untuk mendapatkan nilai Kadar Hambat Minimum50 (KHM50).

Hasil penelitian menunjukkan bahwa jahe emprit dari daerah Jatiyoso, Karanganyar mempunyai aktivitas antioksidan tertinggi dengan IC50 0,150 ± 0,003 mg/g serbuk dan juga kadar flavonoid total tertinggi yaitu 0,0755 ± 0,027 mg/g serbuk. Jahe emprit dari Ambarawa, Semarang memilik kadar fenolik total tertinggi sebesar 1,1069 ± 0,046 mg/g serbuk. Hasil kombinasi FTIR dan kemometrik PCA dapat mengklasifikasikan jahe emprit dari 14 lokasi tumbuh berdasarkan profil metabolit sekundernya. Partial Least Square (PLS) menggunakan spektra FTIR normal dengan bilangan gelombang 3200-850 cm-1 dapat memprediksi kadar gingerol dengan nilai R2 0,9868 untuk kalibrasi dan 0,9735 untuk validasi dengan tingkat kesalahan pada RMSEC dan RMSEP masing-masing sebesar 0,0728 dan 0,123. Minyak atsiri jahe emprit dari Boyolali menunjukkan aktivitas antibakteri tertinggi terhadap S. aureus dengan nilai KHM50 0,2011% v/v, sedang jahe emprit dari Kulonprogo mempunyai aktivitas antibakteri tertinggi terhadap E. coli dengan nilai KHM50 sebesar 0,1893% v/v.

Kombinasi spektra FTIR dan kemometrik dapat mengklasifikasikan jahe emprit dari berbagai lokasi tumbuh berdasarkan profil metabolit sekunder, aktivitas antioksidan, kadar fenolik total, dan flavonoid total. Kombinasi antara kromatogram HPLC dan spektra FTIR dapat memprediksi kadar gingerol jahe emprit, sedangkan kombinasi GC-MS dan OPLS menggunakan software SIMCA dapat menunjukkan pengaruh diskriminatif yang signifikan terhadap senyawa yang berpengaruh pada bakteri S. aureus dan E. coli.

 

 

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Emprit ginger or Zingiber officinale var. Amarum is used by the the community as a traditional medicine with the aim of therapy. The use of herbs is often inconsistent in its effect, due to differences in growing locations that can affect the chemical content of the drug. Therefore, data on chemical content profiles are important as a standardization process. standardization process. One of the efforts in terms of standardization of natural materials is the analysis of their metabolite profiles. analysis of its metabolite profile. This study aims to evaluate the antioxidant activity of of emprit ginger rhizomes, classify emprit ginger powder from different growing locations, correlate the FTIR spectroscopy with chemometric approach, and evaluate the anti-bacterial activity of ginger emprit essential oil.

Ginger powder was tested for free radical scavenging activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, and total flavonoid and total phenolic levels were determined. The powder of ginger emprit was also measured by FTIR spectra combined with HPLC to correlate with gingerol content. Essential oil profiles of ginger emprit from 14 growing sites were obtained using Gas Chromathography-Mass Spectometry (GC-MS) and analyzed by multivariate calibration of Principal Component Analysis (PCA) and Orthogonal Partial Least Squares (OPLS) using SIMCA software. Antibacterial activity of emprit ginger essential oil was performed by microdilution method on Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) bacteria. Analysis of antibacterial activity was determined by probit analysis method to obtain the Minimum Inhibitory Level50 (KHM50) value.

The results showed that emprit ginger from Jatiyoso, Karanganyar had the highest antioxidant activity with IC50 of 0.150 ± 0.003 mg/g powder and also the highest total flavonoid content of 0.0755 ± 0.027 mg/g powder. Emprit ginger from Ambarawa, Semarang has the highest total phenolic content of 1.1069 ± 0.046 mg/g powder. The combination of FTIR and PCA chemometrics can classify emprit ginger from 14 growing locations based on their secondary metabolite profiles. Partial Least Square (PLS) using normal FTIR spectra with wave numbers 3200-850 cm-1 can predict gingerol content with R2 values of 0.9868 for calibration and 0.9735 for validation with error rates in RMSEC and RMSEP of 0.0728 and 0.123, respectively. Emprit ginger essential oil from Boyolali showed the highest antibacterial activity against S. aureus with KHM50 value of 0.2011% v/v, while ginger emprit from Kulonprogo had the highest antibacterial activity against E. coli with KHM50 value of 0.1893% v/v.

The combination of FTIR spectra and chemometrics can classify ginger pith from various growing locations based on secondary metabolite profiles, antioxidant activity, total phenolic content, and total flavonoids. The combination of HPLC chromatograms and FTIR spectra can predict gingerol content of ginger emprit, while the combination of GC-MS and OPLS using SIMCA software can show a significant discriminatory effect on compounds that affect S. aureus and E. coli bacteria.

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Kata Kunci : antibakteri, antioksidan, FTIR, GC-MS, HPLC, rimpang jahe emprit, minyak atsiri

  1. S3-2024-468049-abstract.pdf  
  2. S3-2024-468049-bibliography.pdf  
  3. S3-2024-468049-tableofcontent.pdf  
  4. S3-2024-468049-title.pdf